Journal articles: 'Alginate lyase'

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Yin, Rui, Yan-Jun Yi, Zhuo Chen, Bao-Xun Wang, Xue-Han Li, and Yan-Xia Zhou. "Characterization of a New Biofunctional, Exolytic Alginate Lyase from Tamlana sp. s12 with High Catalytic Activity and Cold-Adapted Features." Marine Drugs 19, no. 4 (March 28, 2021): 191. http://dx.doi.org/10.3390/md19040191.

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Alginate, a major acidic polysaccharide in brown algae, has attracted great attention as a promising carbon source for biorefinery systems. Alginate lyases, especially exo-type alginate lyase, play a critical role in the biorefinery process. Although a large number of alginate lyases have been characterized, few can efficiently degrade alginate comprised of mannuronate (M) and guluronate (G) at low temperatures by means of an exolytic mode. In this study, the gene of a new exo-alginate lyase—Alys1—with high activity (1350 U/mg) was cloned from a marine strain, Tamlana sp. s12. When sodium alginate was used as a substrate, the recombinant enzyme showed optimal activity at 35 °C and pH 7.0–8.0. Noticeably, recombinant Alys1 was unstable at temperatures above 30 °C and had a low melting temperature of 56.0 °C. SDS and EDTA significantly inhibit its activity. These data indicate that Alys1 is a cold-adapted enzyme. Moreover, the enzyme can depolymerize alginates polyM and polyG, and produce a monosaccharide as the minimal alginate oligosaccharide. Primary substrate preference tests and identification of the final oligosaccharide products demonstrated that Alys1 is a bifunctional alginate lyase and prefers M to G. These properties make Alys1 a valuable candidate in both basic research and industrial applications.

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Subaryono, Subaryono, Rosmawaty Peranginangin, Maggy Thenawidjaja Suhartono, and Fransiska Rungkat Zakaria. "Alginate Lyases: Sources, Mechanism of Activity and Potencial Application." Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology 8, no. 3 (December 21, 2013): 105. http://dx.doi.org/10.15578/squalen.v8i3.39.

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Alginate lyases are group of enzymes which catalyze depolymerization of alginate into oligosaccharides. Alginate lyase have been widely used in many applications such as in production of bioactive oligosaccharides, control of polysaccharide rheological properties, and polysaccharide structure analysis. The products of alginate lyase, polysaccharide structure analysis, alginate oligosaccharides (AOS) have many biological activities including act as prebiotics, immune modulator, anticoagulation, antioxidant, anticancer, growth promoting activities, promote production of antibiotics and ethanol. In relation to the importance of alginate lyases, their potential aplications and prospect in development of new bioactive products, we present review of the enzymes, sources, mechanism of activity and potential applications. This paper also discussed some new biological engineering in alginate lyase production.

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Wang, Yanan, Xuehong Chen, Xiaolin Bi, Yining Ren, Qi Han, Yu Zhou, Yantao Han, Ruyong Yao, and Shangyong Li. "Characterization of an Alkaline Alginate Lyase with pH-Stable and Thermo-Tolerance Property." Marine Drugs 17, no. 5 (May 24, 2019): 308. http://dx.doi.org/10.3390/md17050308.

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Alginate oligosaccharides (AOS) show versatile bioactivities. Although various alginate lyases have been characterized, enzymes with special characteristics are still rare. In this study, a polysaccharide lyase family 7 (PL7) alginate lyase-encoding gene, aly08, was cloned from the marine bacterium Vibrio sp. SY01 and expressed in Escherichia coli. The purified alginate lyase Aly08, with a molecular weight of 35 kDa, showed a specific activity of 841 U/mg at its optimal pH (pH 8.35) and temperature (45 °C). Aly08 showed good pH-stability, as it remained more than 80% of its initial activity in a wide pH range (4.0–10.0). Aly08 was also a thermo-tolerant enzyme that recovered 70.8% of its initial activity following heat shock treatment for 5 min. This study also demonstrated that Aly08 is a polyG-preferred enzyme. Furthermore, Aly08 degraded alginates into disaccharides and trisaccharides in an endo-manner. Its thermo-tolerance and pH-stable properties make Aly08 a good candidate for further applications.

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Kam, Natania, Yoo Jung Park, Eun Yeol Lee, and Hee Sook Kim. "Molecular identification of a polyM-specific alginate lyase from Pseudomonas sp. strain KS-408 for degradation of glycosidic linkages between two mannuronates or mannuronate and guluronate in alginate." Canadian Journal of Microbiology 57, no. 12 (December 2011): 1032–41. http://dx.doi.org/10.1139/w11-106.

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An alginate lyase gene of a newly isolated Pseudomonas sp. strain KS-408 was cloned by using PCR with the specific primers designed from homologous nucleotide sequences. A partial protein sequence of KS-408 alginate lyase was homology-modeled on the basis of the crystal structure of A1-III alginate lyase from Sphingomonas sp. strain A1. The proposed 3-D structure of KS-408 alginate lyase shows that Asn-198, His-199, Arg-246, and Tyr-253 residues are conserved for the catalytic active site. The recombinant KS-408-1F (with signal peptide) and KS-408-2F (without signal peptide) alginate lyases with the (His)6 tag consist of 393 (44.5 kDa) and 372 (42.4 kDa) amino acids with isoelectric points of 8.64 and 8.46, respectively. The purified recombinant KS-408 alginate lyase was very stable when it was incubated at 40 °C for 30 min. Alginate oligosaccharides produced by the KS-408-2F alginate lyase were purified on a Bio-Gel P2 column and analyzed by thin-layer chromatography, fast-protein liquid chromatography, and electrospray ionization mass spectrometry. 1H NMR data showed that the KS-408-2F alginate lyase cleaved the glycosidic linkages between two mannuronates (mannuronate-β(1–4)-mannuronate) or mannuronate and guluronate (mannuronate-β(1–4)-guluronate), indicating that the KS-408 alginate lyase is a polyM-specific lyase.

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Zhuang, Jingjing, Keke Zhang, Xiaohua Liu, Weizhi Liu, Qianqian Lyu, and Aiguo Ji. "Characterization of a Novel PolyM-Preferred Alginate Lyase from Marine Vibrio splendidus OU02." Marine Drugs 16, no. 9 (August 22, 2018): 295. http://dx.doi.org/10.3390/md16090295.

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Alginate lyases are enzymes that degrade alginate into oligosaccharides which possess a variety of biological activities. Discovering and characterizing novel alginate lyases has great significance for industrial and medical applications. In this study, we reported a novel alginate lyase, AlyA-OU02, derived from the marine Vibrio splendidus OU02. The BLASTP searches showed that AlyA-OU02 belonged to polysaccharide lyase family 7 (PL7) and contained two consecutive PL7 domains, which was rare among the alginate lyases in PL7 family. Both the two domains, AlyAa and AlyAb, had lyase activities, while AlyAa exhibited polyM preference, and AlyAb was polyG-preferred. In addition, the enzyme activity of AlyAa was much higher than AlyAb at 25 °C. The full-length enzyme of AlyA-OU02 showed polyM preference, which was the same as AlyAa. AlyAa degraded alginate into di-, tri-, and tetra-alginate oligosaccharides, while AlyAb degraded alginate into tri-, tetra-, and penta-alginate oligosaccharides. The degraded products of AlyA-OU02 were similar to AlyAa. Our work provided a potential candidate in the application of alginate oligosaccharide production and the characterization of the two domains might provide insights into the use of alginate of this organism.

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Badur, Ahmet H., Sujit Sadashiv Jagtap, Geethika Yalamanchili, Jung-Kul Lee, Huimin Zhao, and Christopher V. Rao. "Alginate Lyases from Alginate-Degrading Vibrio splendidus 12B01 Are Endolytic." Applied and Environmental Microbiology 81, no. 5 (January 2, 2015): 1865–73. http://dx.doi.org/10.1128/aem.03460-14.

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ABSTRACTAlginate lyases are enzymes that degrade alginate through β-elimination of the glycosidic bond into smaller oligomers. We investigated the alginate lyases fromVibrio splendidus12B01, a marine bacterioplankton species that can grow on alginate as its sole carbon source. We identified, purified, and characterized four polysaccharide lyase family 7 alginates lyases, AlyA, AlyB, AlyD, and AlyE, fromV. splendidus12B01. The four lyases were found to have optimal activity between pH 7.5 and 8.5 and at 20 to 25°C, consistent with their use in a marine environment. AlyA, AlyB, AlyD, and AlyE were found to exhibit a turnover number (kcat) for alginate of 0.60 ± 0.02 s−1, 3.7 ± 0.3 s−1, 4.5 ± 0.5 s−1, and 7.1 ± 0.2 s−1, respectively. TheKmvalues of AlyA, AlyB, AlyD, and AlyE toward alginate were 36 ± 7 μM, 22 ± 5 μM, 60 ± 2 μM, and 123 ± 6 μM, respectively. AlyA and AlyB were found principally to cleave the β-1,4 bonds between β-d-mannuronate and α-l-guluronate and subunits; AlyD and AlyE were found to principally cleave the α-1,4 bonds involving α-l-guluronate subunits. The four alginate lyases degrade alginate into longer chains of oligomers.

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Li, Qian, Fu Hu, Benwei Zhu, Yun Sun, and Zhong Yao. "Biochemical Characterization and Elucidation of Action Pattern of a Novel Polysaccharide Lyase 6 Family Alginate Lyase from Marine Bacterium Flammeovirga sp. NJ-04." Marine Drugs 17, no. 6 (May 31, 2019): 323. http://dx.doi.org/10.3390/md17060323.

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Alginate lyases have been widely used to prepare alginate oligosaccharides in food, agricultural, and medical industries. Therefore, discovering and characterizing novel alginate lyases with excellent properties has drawn increasing attention. Herein, a novel alginate lyase FsAlyPL6 of Polysaccharide Lyase (PL) 6 family is identified and biochemically characterized from Flammeovirga sp. NJ-04. It shows highest activity at 45 °C and could retain 50% of activity after being incubated at 45 °C for 1 h. The Thin-Layer Chromatography (TLC) and Electrospray Ionization Mass Spectrometry (ESI-MS) analysis indicates that FsAlyPL6 endolytically degrades alginate polysaccharide into oligosaccharides ranging from monosaccharides to pentasaccharides. In addition, the action pattern of the enzyme is also elucidated and the result suggests that FsAlyPL6 could recognize tetrasaccharide as the minimal substrate and cleave the glycosidic bonds between the subsites of −1 and +3. The research provides extended insights into the substrate recognition and degradation pattern of PL6 alginate lyases, which may further expand the application of alginate lyases.

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Pilgaard, Bo, Marlene Vuillemin, Jesper Holck, Casper Wilkens, and Anne S. Meyer. "Specificities and Synergistic Actions of Novel PL8 and PL7 Alginate Lyases from the Marine Fungus Paradendryphiella salina." Journal of Fungi 7, no. 2 (January 25, 2021): 80. http://dx.doi.org/10.3390/jof7020080.

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Alginate is an anionic polysaccharide abundantly present in the cell walls of brown macroalgae. The enzymatic depolymerization is performed solely by alginate lyases (EC 4.2.2.x), categorized as polysaccharide lyases (PLs) belonging to 12 different PL families. Until now, the vast majority of the alginate lyases have been found in bacteria. We report here the first extensive characterization of four alginate lyases from a marine fungus, the ascomycete Paradendryphiella salina, a known saprophyte of seaweeds. We have identified four polysaccharide lyase encoding genes bioinformatically in P. salina, one PL8 (PsMan8A), and three PL7 alginate lyases (PsAlg7A, -B, and -C). PsMan8A was demonstrated to exert exo-action on polymannuronic acid, and no action on alginate, indicating that this enzyme is most likely an exo-acting polymannuronic acid specific lyase. This enzyme is the first alginate lyase assigned to PL8 and polymannuronic acid thus represents a new substrate specificity in this family. The PL7 lyases (PsAlg7A, -B, and -C) were found to be endo-acting alginate lyases with different activity optima, substrate affinities, and product profiles. PsAlg7A and PsMan8A showed a clear synergistic action for the complete depolymerization of polyM at pH 5. PsAlg7A depolymerized polyM to mainly DP5 and DP3 oligomers and PsMan8A to dimers and monosaccharides. PsAlg7B and PsAlg7C showed substrate affinities towards both polyM and polyG at pH 8, depolymerizing both substrates to DP9-DP2 oligomers. The findings elucidate how P. salina accomplishes alginate depolymerization and provide insight into an efficient synergistic cooperation that may provide a new foundation for enzyme selection for alginate degradation in seaweed bioprocessing.

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Wei, Wei, Xin Zhang, Zhaozhi Hou, Xinyu Hu, Yuan Wang, Caizheng Wang, Shujing Yang, Henglin Cui, and Lin Zhu. "Microbial Regulation of Deterioration and Preservation of Salted Kelp under Different Temperature and Salinity Conditions." Foods 10, no. 8 (July 26, 2021): 1723. http://dx.doi.org/10.3390/foods10081723.

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High salinity is an effective measure to preserve kelp, but salted kelp can still deteriorate after long-term preservation. In order to clarify the key conditions and microbial behavior of salted kelp preservation, 10% (S10), 20% (S20), and 30% (S30) salt concentrations were evaluated at 25 °C (T25) and 4 °C (T4). After 30 days storage, these salted kelps showed different states including rot (T25S10), softening (T25S20), and undamaged (other samples). By detecting polysaccharide lyase activity and performing high-throughput sequencing of the prokaryotic 16S rRNA sequence and metagenome, we found that deteriorated kelps (T25S10 and T25S20) had significantly higher alginate lyase activity and bacterial relative abundance than other undamaged samples. Dyella, Saccharophagus, Halomonas, Aromatoleum, Ulvibacter, Rhodopirellula, and Microbulbifer were annotated with genes encoding endonuclease-type alginate lyases, while Bacillus and Thiobacillus were annotated as the exonuclease type. Additionally, no alginate lyase activity was detected in undamaged kelps, whose dominant microorganisms were halophilic archaea without alginate lyase-encoding genes. These results indicated that room-temperature storage may promote salted kelp deterioration due to the secretion of bacterial alginate lyase, while ultra-high-salinity and low-temperature storage can inhibit bacterial alginate lyase and promote the growth of halophilic archaea without alginate lyase, thus achieving the preservation of salted kelp.

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Xu, Fei, Xiu-Lan Chen, Xiao-Hui Sun, Fang Dong, Chun-Yang Li, Ping-Yi Li, Haitao Ding, Yin Chen, Yu-Zhong Zhang, and Peng Wang. "Structural and molecular basis for the substrate positioning mechanism of a new PL7 subfamily alginate lyase from the arctic." Journal of Biological Chemistry 295, no. 48 (September 23, 2020): 16380–92. http://dx.doi.org/10.1074/jbc.ra120.015106.

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Alginate lyases play important roles in alginate degradation in the ocean. Although a large number of alginate lyases have been characterized, little is yet known about those in extremely cold polar environments, which may have unique mechanisms for environmental adaptation and for alginate degradation. Here, we report the characterization of a novel PL7 alginate lyase AlyC3 from Psychromonas sp. C-3 isolated from the Arctic brown alga Laminaria, including its phylogenetic classification, catalytic properties, and structure. We propose the establishment of a new PM-specific subfamily of PL7 (subfamily 6) represented by AlyC3 based on phylogenetic analysis and enzymatic properties. Structural and biochemical analyses showed that AlyC3 is a dimer, representing the first dimeric endo-alginate lyase structure. AlyC3 is activated by NaCl and adopts a novel salt-activated mechanism; that is, salinity adjusts the enzymatic activity by affecting its aggregation states. We further solved the structure of an inactive mutant H127A/Y244A in complex with a dimannuronate molecule and proposed the catalytic process of AlyC3 based on structural and biochemical analyses. We show that Arg82 and Tyr190 at the two ends of the catalytic canyon help the positioning of the repeated units of the substrate and that His127, Tyr244, Arg78, and Gln125 mediate the catalytic reaction. Our study uncovers, for the first time, the amino acid residues for alginate positioning in an alginate lyase and demonstrates that such residues involved in alginate positioning are conserved in other alginate lyases. This study provides a better understanding of the mechanisms of alginate degradation by alginate lyases.

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Zhang, Zhelun, Luyao Tang, Mengmeng Bao, Zhigang Liu, Wengong Yu, and Feng Han. "Functional Characterization of Carbohydrate-Binding Modules in a New Alginate Lyase, TsAly7B, from Thalassomonas sp. LD5." Marine Drugs 18, no. 1 (December 26, 2019): 25. http://dx.doi.org/10.3390/md18010025.

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Alginate lyases degrade alginate into oligosaccharides, of which the biological activities have vital roles in various fields. Some alginate lyases contain one or more carbohydrate-binding modules (CBMs), which assist the function of the catalytic modules. However, the precise function of CBMs in alginate lyases has yet to be fully elucidated. We have identified a new multi-domain alginate lyase, TsAly7B, in the marine bacterium Thalassomonas sp. LD5. This novel lyase contains an N-terminal CBM9, an internal CBM32, and a C-terminal polysaccharide lyase family 7 (PL7) catalytic module. To investigate the specific function of each of these CBMs, we expressed and characterized the full-length TsAly7B and three truncated mutants: TM1 (CBM32-PL7), TM2 (CBM9-PL7), and TM3 (PL7 catalytic module). CBM9 and CBM32 could enhance the degradation of alginate. Notably, the specific activity of TM2 was 7.6-fold higher than that of TM3. CBM32 enhanced the resistance of the catalytic module to high temperatures. In addition, a combination of CBM9 and CBM32 showed enhanced thermostability when incubated at 80 °C for 1 h. This is the first report that finds CBM9 can significantly improve the ability of enzyme degradation. Our findings provide new insight into the interrelationships of tandem CBMs and alginate lyases and other polysaccharide-degrading enzymes, which may inspire CBM fusion strategies.

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Zhang, Yong-Hui, Yuan Shao, Chao Jiao, Qiu-Ming Yang, Hui-Fen Weng, and An-Feng Xiao. "Characterization and Application of an Alginate Lyase, Aly1281 from Marine Bacterium Pseudoalteromonas carrageenovora ASY5." Marine Drugs 18, no. 2 (January 31, 2020): 95. http://dx.doi.org/10.3390/md18020095.

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Alginate extracted from widely cultured brown seaweed can be hydrolyzed by alginate lyase to produce alginate oligosaccharides (AOS) with intriguing biological activities. Herein, a novel alginate lyase Aly1281 was cloned from marine bacterium Pseudoalteromonas carrageenovora ASY5 isolated from mangrove soil and found to belong to polysaccharide lyase family 7. Aly1281 exhibited maximum activity at pH 8.0 and 50 °C and have broad substrate specificity for polyguluronate and polymannuronate. Compared with other alginate lyases, Aly1281 exhibited high degradation specificity and mainly produced di-alginate oligosaccharides which displayed good antioxidant function to reduce ferric and scavenge radicals such as hydroxyl, ABTS+ and DPPH. Moreover, the catalytic activity and kinetic performance of Aly1281 were highly improved with the addition of salt, demonstrating a salt-activation property. A putative conformational structural feature of Aly1281 was found by MD simulation analysis for understanding the salt-activation effect.

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Afni, Fahriza Sri, Sri Purwaningsih, Mala Nurilmala, and Rosmawati Peranginangin. "Production of Alginate Oligosaccharides (AOS) as Prebiotic Ingredients through by Alginate lyase enzyme." Jurnal Pengolahan Hasil Perikanan Indonesia 20, no. 1 (April 28, 2017): 109. http://dx.doi.org/10.17844/jphpi.v20i1.16498.

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Prebiotics is indigestible foods that can not be digested but can stimulate the growth and activity of bacteria in the digestive tract effecting human health. Alginate oligosaccharides (AOS) can be used as a source of prebiotic. That compounds can be produced enzymatically by cutting long chain alginates using alginate lyase. The aim of this study was to produce alginate lyase enzyme then producing Alginate oligosaccharides (AOS) as a prebiotic ingredients. The alginate lyase enzyme can be produced from Bacillus megaterium bacteria using a discontinuous fermentor. The enzyme was optimum temperature of 45°C and an optimum pH of 7.0. Alginate oligosaccharides production was performed with the addition of different enzyme concentrations 25, 50, 75, and 100 U. The result of the addition of enzyme (25, 50,75 U) showed that the value of polymerization degrees (DP) were between 4-5. However, the addition of enzyme (100 U) was in the range of DP 3-4. Bacterial probiotic growth test results of Bifidobacteria and Lactobacillus showed that 1% added AOS media were able to increase the growth of probiotic bacteria compared to the media without addition of AOS. The addition Alginate lyase activity of 50 U in AOS production is the best treatment of both probiotic bacteria.

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Preston, Lori A., T. Y. Wong, Carol L. Bender, and Neal L. Schiller. "Characterization of Alginate Lyase from Pseudomonas syringae pv. syringae." Journal of Bacteriology 182, no. 21 (November 1, 2000): 6268–71. http://dx.doi.org/10.1128/jb.182.21.6268-6271.2000.

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ABSTRACT The gene encoding alginate lyase (algL) inPseudomonas syringae pv. syringae was cloned, sequenced, and overexpressed in Escherichia coli. Alginate lyase activity was optimal when the pH was 7.0 and when assays were conducted at 42°C in the presence of 0.2 M NaCl. In substrate specificity studies, AlgL from P. syringae showed a preference for deacetylated polymannuronic acid. Sequence alignment with other alginate lyases revealed conserved regions within AlgL likely to be important for the structure and/or function of the enzyme. Site-directed mutagenesis of histidine and tryptophan residues at positions 204 and 207, respectively, indicated that these amino acids are critical for lyase activity.

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Zhang, Keke, Tao Liu, Weizhi Liu, and Qianqian Lyu. "Structural insights into the substrate-binding cleft of AlyF reveal the first long-chain alginate-binding mode." Acta Crystallographica Section D Structural Biology 77, no. 3 (February 17, 2021): 336–46. http://dx.doi.org/10.1107/s205979832100005x.

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The products of alginate degradation, alginate oligosaccharides (AOS), have potential applications in many areas, including functional foods and marine drugs. Enzyme-based approaches using alginate lyases have advantages in the preparation of well defined AOS and have attracted much attention in recent years. However, a lack of structural insight into the whole substrate-binding cleft for most known alginate lyases severely hampers their application in the industrial generation of well defined AOS. To solve this issue, AlyF was co-crystallized with the long alginate oligosaccharide G6 (L-hexaguluronic acid hexasodium salt), which is the longest bound substrate in all solved alginate lyase complex structures. AlyF formed interactions with G6 from subsites −3 to +3 without additional substrate-binding site interactions, suggesting that the substrate-binding cleft of AlyF was fully occupied by six sugars, which was further confirmed by isothermal titration calorimetry and differential scanning calorimetry analyses. More importantly, a combination of structural comparisons and mutagenetic analyses determined that three key loops (loop 1, Lys215–Glu236; loop 2, Gln402–Ile416; loop 3, Arg334–Gly348) mainly function in binding long substrates (degree of polymerization of >4). The potential flexibility of loop 1 and loop 2 might enable the substrate to continue to enter the cleft after binding to subsites +1 to +3; loop 3 stabilizes and orients the substrate at subsites −2 and −3. Taken together, these results provide the first possible alginate lyase–substrate binding profile for long-chain alginates, facilitating the rational design of new enzymes for industrial purposes.

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Duan, Gaofei, Feng Han, and Wengong Yu. "Cloning, sequence analysis, and expression of gene alyPI encoding an alginate lyase from marine bacterium Pseudoalteromonas sp. CY24." Canadian Journal of Microbiology 55, no. 9 (September 2009): 1113–18. http://dx.doi.org/10.1139/w09-051.

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The alginate lyase encoding gene (alyPI) of marine bacterium Pseudoalteromonas sp. CY24 was cloned using a battery of PCR techniques. Gene alyPI was composed of a 1575 bp open reading frame encoding a protein of 57.4 kDa containing 524 amino acid residues with a signal peptide of 23 amino acids. The AlyPI protein was expressed in Escherichia coli with a His-tag sequence fused at the C-terminal end and purified to electrophoretic homogeneity using Ni-sepharose affinity chromatography. AlyPI was most active at 40 °C and pH 7.0 in the presencce of 0.1 mol/L NaCl and stable over a broad range of pH, 6.0–10.6. The presence of Na+, K+, Mn2+, Ca2+, and Fe3+ can enhance the enzyme activity. The alginate lyase consensus region YFKAGXYXQ, regarded as a striking feature at the C termini of several alginate lyase of ~30 kDa, was found in AlyPI, which belongs to the ~60 kDa group. Another nine amino acid consensus region, YXRSELREM, only found in G-specific alginate lyases previously existed in AlyPI, which could degrade sodium alginate, M blocks, and G blocks and appeared to be a broad substrate-specific alginate lyase.

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Subaryono, Subaryono, Rosmawati Perangiangin, Maggy Thenawidjaja Suhartono, and Fransiska Rungkat Zakaria. "Imunomodulator Activity of Alginate Oligosaccharides from Alginate Sargassum crassifolium." Jurnal Pengolahan Hasil Perikanan Indonesia 20, no. 1 (April 25, 2017): 63. http://dx.doi.org/10.17844/jphpi.v20i1.16434.

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Alginate oligosaccharides (AOS) are oligosaccharides produced from depolimerization of the alginate polymer, and is reported to have various biological activities. The study aims is to determine the effect of AOS production conditions and their effects on products and its activities as an immunomodulatory compound. Production of alginate oligosaccharides (AOS) enzymatically carried out with the help of alginate lyase enzyme produced from the bacterium Bacillus megaterium S245. Variation of incubation time is 2, 4, 6 and 8 hours at concentrations of alginate lyase enzyme addition of 25, 50, 75 and 100U. Treatment of enzyme concentration and the duration of incubation in the production of AOS produces a degree of polymerization (DP) 2-7. In vitro activity test showed AOS is have ability to induce cell proliferation of human lymphocytes. This type of cell lymphocytes proliferation induced by AOS is a CD 8 cells or cytotoxic T cell and non cell CD4 / CD8. AOS production conditions with the addition of alginate lyase enzyme 50 U and incubation period 2 hours has produce AOS with the highest index of lymphocyte proliferation 117.6+3.6% or an increase of 43.24% compared to the native alginat polymer.

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Manns, D., C. Nyffenegger, B. Saake, and A. S. Meyer. "Impact of different alginate lyases on combined cellulase–lyase saccharification of brown seaweed." RSC Advances 6, no. 51 (2016): 45392–401. http://dx.doi.org/10.1039/c6ra06669k.

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Alginate attack characteristics and impact on cellulase–lyase catalyzed saccharification of brown seaweed were compared for three microbial PL7 alginate lyases (EC 4.2.2.-) two of them heterologously expressed in Escherichia coli as part of the work.

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Hashimoto, Wataru, Osamu Miyake, Keiko Momma, Shigeyuki Kawai, and Kousaku Murata. "Molecular Identification of Oligoalginate Lyase ofSphingomonas sp. Strain A1 as One of the Enzymes Required for Complete Depolymerization of Alginate." Journal of Bacteriology 182, no. 16 (August 15, 2000): 4572–77. http://dx.doi.org/10.1128/jb.182.16.4572-4577.2000.

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ABSTRACT A bacterium, Sphingomonas sp. strain A1, can incorporate alginate into cells through a novel ABC (ATP-binding cassette) transporter system specific to the macromolecule. The transported alginate is depolymerized to di- and trisaccharides by three kinds of cytoplasmic alginate lyases (A1-I [66 kDa], A1-II [25 kDa], and A1-III [40 kDa]) generated from a single precursor through posttranslational autoprocessing. The resultant alginate oligosaccharides were degraded to monosaccharides by cytoplasmic oligoalginate lyase. The enzyme and its gene were isolated from the bacterial cells grown in the presence of alginate. The purified enzyme was a monomer with a molecular mass of 85 kDa and cleaved glycosidic bonds not only in oligosaccharides produced from alginate by alginate lyases but also in polysaccharides (alginate, polymannuronate, and polyguluronate) most efficiently at pH 8.0 and 37°C. The reaction catalyzed by the oligoalginate lyase was exolytic and thought to play an important role in the complete depolymerization of alginate in Sphingomonas sp. strain A1. The gene for this novel enzyme consisted of an open reading frame of 2,286 bp encoding a polypeptide with a molecular weight of 86,543 and was located downstream of the genes coding for the precursor of alginate lyases (aly) and the ABC transporter (algS,algM1, and algM2). This result indicates that the genes for proteins required for the transport and complete depolymerization of alginate are assembled to form a cluster.

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Lamppa, John W., and Karl E. Griswold. "Alginate Lyase Exhibits Catalysis-Independent Biofilm Dispersion and Antibiotic Synergy." Antimicrobial Agents and Chemotherapy 57, no. 1 (October 15, 2012): 137–45. http://dx.doi.org/10.1128/aac.01789-12.

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ABSTRACTMore than 2 decades of study support the hypothesis that alginate lyases are promising therapeutic candidates for treating mucoidPseudomonas aeruginosainfections. In particular, the enzymes' ability to degrade alginate, a key component of mucoid biofilm matrix, has been the presumed mechanism by which they disrupt biofilms and enhance antibiotic efficacy. The systematic studies reported here show that, in anin vitromodel, alginate lyase dispersion ofP. aeruginosabiofilms and enzyme synergy with tobramycin are completely decoupled from catalytic activity. In fact, equivalent antibiofilm effects can be achieved with bovine serum albumin or simple amino acids. These results provide new insights into potential mechanisms of alginate lyase therapeutic activity, and they should motivate a careful reexamination of the fundamental assumptions underlying interest in enzymatic biofilm dispersion.

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Daboor, Said M., Renee Raudonis, Alejandro Cohen, John R. Rohde, and Zhenyu Cheng. "Marine Bacteria, A Source for Alginolytic Enzyme to Disrupt Pseudomonas aeruginosa Biofilms." Marine Drugs 17, no. 5 (May 24, 2019): 307. http://dx.doi.org/10.3390/md17050307.

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Pseudomonas aeruginosa biofilms are typically associated with the chronic lung infection of cystic fibrosis (CF) patients and represent a major challenge for treatment. This opportunistic bacterial pathogen secretes alginate, a polysaccharide that is one of the main components of its biofilm. Targeting this major biofilm component has emerged as a tempting therapeutic strategy for tackling biofilm-associated bacterial infections. The enormous potential in genetic diversity of the marine microbial community make it a valuable resource for mining activities responsible for a broad range of metabolic processes, including the alginolytic activity responsible for degrading alginate. A collection of 36 bacterial isolates were purified from marine water based on their alginolytic activity. These isolates were identified based on their 16S rRNA gene sequences. Pseudoalteromonas sp. 1400 showed the highest alginolytic activity and was further confirmed to produce the enzyme alginate lyase. The purified alginate lyase (AlyP1400) produced by Pseudoalteromonas sp. 1400 showed a band of 23 KDa on a protein electrophoresis gel and exhibited a bifunctional lyase activity for both poly-mannuronic acid and poly-glucuronic acid degradation. A tryptic digestion of this gel band analyzed by liquid chromatography-tandem mass spectrometry confirmed high similarity to the alginate lyases in polysaccharide lyase family 18. The purified alginate lyase showed a maximum relative activity at 30 °C at a slightly acidic condition. It decreased the sodium alginate viscosity by over 90% and reduced the P. aeruginosa (strain PA14) biofilms by 69% after 24 h of incubation. The combined activity of AlyP1400 with carbenicillin or ciprofloxacin reduced the P. aeruginosa biofilm thickness, biovolume and surface area in a flow cell system. The present data revealed that AlyP1400 combined with conventional antibiotics helped to disrupt the biofilms produced by P. aeruginosa and can be used as a promising combinational therapeutic strategy.

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22

ZILDA, DEWI SESWITA, YULIYANTI YULIANTI, RIZKY FAUZIYAH SHOLIHAH, SUBARYONO SUBARYONO, YUSRO NURI FAWZYA, and HARI EKO IRIANTO. "A novel Bacillus sp. isolated from rotten seaweed: Identification and characterization alginate lyase it is produced." Biodiversitas Journal of Biological Diversity 20, no. 4 (March 30, 2019): 1166–72. http://dx.doi.org/10.13057/biodiv/d200432.

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Abstract. Zilda DS, Yulianti Y, Sholihah RF, Subaryono S, Fawzya YN, Irianto HE.2019. A novel Bacillus sp. isolated from rotten seaweed: identification and characterization alginate lyase it is produced. Biodiversitas 20: 1166-1172. Alginate lyase has been known as potential biocatalyst not only for industrial but also medicinal application especially for the production of oligosaccharides which have distinct bioactivities. An alginate lyase, AlgT513, has been isolated from rotten seaweed bacterium strain T513 and characterized. The bacterium showed low similarity (95%) with Bacillus tequilensis strain 10b based on 16S rDNA sequence indicating that Alg07 may be a novel Bacillus species. The bacterium forms a clear zone on solid medium with 0.5% sodium alginate addition. The optimum temperature and pH were 50ºC and 8 respectively. AlgT513 maintained stability at board pHs of 4-9 and temperature of 45ºC. Metal ions Mg2+, Ca2+ and K+ increase the activity of the enzyme while Zn2+, Co2+ and Li+ strongly inhibit it. NaCl inhibits AlgT513 activity where most of the alginate lyases need it to reach maximum activity. AlgT513 is suggested as a serine metalloenzyme due to inhibition of ethylenediaminetetraacetic acid (EDTA) and phenylmethylsulfonyl fluoride (PMSF).

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Ma, Yan, Jie Li, Xin-Yue Zhang, Hao-Dong Ni, Feng-Biao Wang, Hai-Ying Wang, and Zhi-Peng Wang. "Characterization of a New Intracellular Alginate Lyase with Metal Ions-Tolerant and pH-Stable Properties." Marine Drugs 18, no. 8 (August 9, 2020): 416. http://dx.doi.org/10.3390/md18080416.

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Alginate lyases play an important role in alginate oligosaccharides (AOS) preparation and brown seaweed processing. Many extracellular alginate lyases have been characterized to develop efficient degradation tools needed for industrial applications. However, few studies focusing on intracellular alginate lyases have been conducted. In this work, a novel intracellular alkaline alginate lyase Alyw202 from Vibrio sp. W2 was cloned, expressed and characterized. Secretory expression was performed in a food-grade host, Yarrowia lipolytica. Recombinant Alyw202 with a molecular weight of approximately 38.3 kDa exhibited the highest activity at 45 °C and more than 60% of the activity in a broad pH range of 3.0 to 10.0. Furthermore, Alyw202 showed remarkable metal ion-tolerance, NaCl independence and the capacity of degrading alginate into oligosaccharides of DP2-DP4. Due to the unique pH-stable and high salt-tolerant properties, Alyw202 has potential applications in the food and pharmaceutical industries.

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Hisano, Tomohiro, Minoru Nishimura, Yoshimasa Yonemoto, Shiro Abe, Tetsuo Yamashita, Kenji Sakaguchi, Akira Kimura, and Kousaku Murata. "Bacterial alginate lyase highly active on acetylated alginates." Journal of Fermentation and Bioengineering 75, no. 5 (January 1993): 332–35. http://dx.doi.org/10.1016/0922-338x(93)90129-v.

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25

Zhu, Benwei, Limin Ning, Yucui Jiang, and Lin Ge. "Biochemical Characterization and Degradation Pattern of a Novel Endo-Type Bifunctional Alginate Lyase AlyA from Marine Bacterium Isoptericola halotolerans." Marine Drugs 16, no. 8 (July 31, 2018): 258. http://dx.doi.org/10.3390/md16080258.

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Alginate lyases are important tools to prepare oligosaccharides with various physiological activities by degrading alginate. Particularly, the bifunctional alginate lyase can efficiently hydrolyze the polysaccharide into oligosaccharides. Herein, we cloned and identified a novel bifunctional alginate lyase, AlyA, with a high activity and broad substrate specificity from bacterium Isoptericola halotolerans NJ-05 for oligosaccharides preparation. For further applications in industry, the enzyme has been characterized and its action mode has been also elucidated. It exhibited the highest activity (7984.82 U/mg) at pH 7.5 and 55 °C. Additionally, it possessed a broad substrate specificity, showing high activities towards not only polyM (polyβ-d-mannuronate) (7658.63 U/mg), but also polyG (poly α-l-guluronate) (8643.29 U/mg). Furthermore, the Km value of AlyA towards polyG (3.2 mM) was lower than that towards sodium alginate (5.6 mM) and polyM (6.7 mM). TLC (Thin Layer Chromatography) and ESI-MS (Electrospray Ionization Mass Spectrometry) were used to study the action mode of the enzyme, showing that it can hydrolyze the substrates in an endolytic manner to release a series of oligosaccharides such as disaccharide, trisaccharide, and tetrasaccharide. This study provided extended insights into the substrate recognition and degrading pattern of the alginate lyases, with a broad substrate specificity.

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Younis, R. W. "PRODUCTION, PURIFICATION AND INHIBITION OF ALGINATE LY-ASE FROM LOCAL ISOLATE OF PSEUDOMONAS AERUGINOSA NA11." IRAQI JOURNAL OF AGRICULTURAL SCIENCES 51, no. 6 (December 23, 2020): 1726–39. http://dx.doi.org/10.36103/ijas.v51i6.1201.

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The point of this study was for determine of the optimum conditions and purification of alginate lyase from local isolate of Pseudomonas aeruginosa NA11, and inhibition of enzyme by various plant extracts. Forty local isolate s of pathogenic Pseudomonas aeruginosa were screened for their ability to produce alginate lyase. Local isolate of P.aeruginosa NA11 showed the maximum efficiency for produce of alginate lyase with high specific activity (14.4 U/mg). Several factors that influence on alginate lyase production from local isolate of P.aeruginosa NA11 were studied, these factors included the type of media, carbon source, nitrogen source, the incubation temperature, pH, and the incubation period. The highest yield of alginate lyase was obtained with the A medium supplemented with 0.5 % of glucose and sodium nitrate at pH 7.5 after 24 hr. incubation at 37 °C. Two chromatographic techniques were used for purification of alginate lyase after precipitation by ammonium sulphate with saturated ratio (0-70 %), including, ion exchange chromatography by DEAE-cellulose and gel filtration by Sephadex G-100. The two steps gave the specific activity of 155.8 U/mg protein, the purification fold was 4.15 and enzymatic yield was 64 %. The molecular weight of partial purified alginate lyase was 57 KDa. The results of antioxidant activity tests for different plants extracts utilizing DPPH radical scavenging activity were showed that the saad extract has high antioxidant activity (71 %) more than other plants extracts. While the results for inhibition experiment of alginate lyase were demonstrated that saad extract was the best inhibitor with inhibition ratio of 86 %.

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Han, Wenjun, Jingyan Gu, Yuanyuan Cheng, Huihui Liu, Yuezhong Li, and Fuchuan Li. "Novel Alginate Lyase (Aly5) from a Polysaccharide-Degrading Marine Bacterium, Flammeovirga sp. Strain MY04: Effects of Module Truncation on Biochemical Characteristics, Alginate Degradation Patterns, and Oligosaccharide-Yielding Properties." Applied and Environmental Microbiology 82, no. 1 (October 30, 2015): 364–74. http://dx.doi.org/10.1128/aem.03022-15.

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ABSTRACTAlginate lyases are important tools for oligosaccharide preparation, medical treatment, and energy bioconversion. Numerous alginate lyases have been elucidated. However, relatively little is known about their substrate degradation patterns and product-yielding properties, which is a limit to wider enzymatic applications and further enzyme improvements. Herein, we report the characterization and module truncation of Aly5, the first alginate lyase obtained from the polysaccharide-degrading bacteriumFlammeovirga. Aly5 is a 566-amino-acid protein and belongs to a novel branch of the polysaccharide lyase 7 (PL7) superfamily. The protein rAly5 is an endolytic enzyme of alginate and associated oligosaccharides. It prefers guluronate (G) to mannuronate (M). Its smallest substrate is an unsaturated pentasaccharide, and its minimum product is an unsaturated disaccharide. The final alginate digests contain unsaturated oligosaccharides that generally range from disaccharides to heptasaccharides, with the tetrasaccharide fraction constituting the highest mass concentration. The disaccharide products are identified as ΔG units. While interestingly, the tri- and tetrasaccharide fractions each contain higher proportions of ΔG to ΔM ends, the larger final products contain only ΔM ends, which constitute a novel oligosaccharide-yielding property of guluronate lyases. The deletion of the noncatalytic region of Aly5 does not alter its M/G preference but significantly decreases the enzymatic activity and enzyme stability. Notably, the truncated protein accumulates large final oligosaccharide products but yields fewer small final products than Aly5, which are codetermined by its M/G preference to and size enlargement of degradable oligosaccharides. This study provides novel enzymatic properties and catalytic mechanisms of a guluronate lyase for potential uses and improvements.

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Gimmestad, Martin, Helga Ertesvåg, Tonje Marita Bjerkan Heggeset, Olav Aarstad, Britt Iren Glærum Svanem, and Svein Valla. "Characterization of Three New Azotobacter vinelandii Alginate Lyases, One of Which Is Involved in Cyst Germination." Journal of Bacteriology 191, no. 15 (May 26, 2009): 4845–53. http://dx.doi.org/10.1128/jb.00455-09.

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ABSTRACT Alginates are polysaccharides composed of 1-4-linked β-d-mannuronic acid and α-l-guluronic acid. The polymer can be degraded by alginate lyases, which cleave the polysaccharide using a β-elimination reaction. Two such lyases have previously been identified in the soil bacterium Azotobacter vinelandii, as follows: the periplasmic AlgL and the secreted bifunctional mannuronan C-5 epimerase and alginate lyase AlgE7. In this work, we describe the properties of three new lyases from this bacterium, AlyA1, AlyA2, and AlyA3, all of which belong to the PL7 family of polysaccharide lyases. One of the enzymes, AlyA3, also contains a C-terminal module similar to those of proteins secreted by a type I secretion system, and its activity is stimulated by Ca2+. All three enzymes preferably cleave the bond between guluronic acid and mannuronic acid, resulting in a guluronic acid residue at the new reducing end, but AlyA3 also degrades the other three possible bonds in alginate. Strains containing interrupted versions of alyA1, alyA3, and algE7 were constructed, and their phenotypes were analyzed. Genetically pure alyA2 mutants were not obtained, suggesting that this gene product may be important for the bacterium during vegetative growth. After centrifugation, cultures from the algE7 mutants form a large pellet containing alginate, indicating that AlgE7 is involved in the release of alginate from the cells. Upon encountering adverse growth conditions, A. vinelandii will form a resting stage called cyst. Alginate is a necessary part of the protective cyst coat, and we show here that strains lacking alyA3 germinate poorly compared to wild-type cells.

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Subaryono, Subaryono, Yuwanita Ardilasari, Rosmawaty Peranginangin, Fransisca Rungkat Zakaria, and Maggy Thenawidjaja Suhartono. "Alginate Lyase from Indonesian Bacillus megaterium S245 Shows Activities Toward Polymannuronate and Polyguluronate." Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology 11, no. 2 (February 28, 2017): 45. http://dx.doi.org/10.15578/squalen.v11i2.250.

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Screening of alginate lyase producing bacteria associated with seaweed Sargassum crassifolium was carried out, and isolate S245, identified as Bacillus megaterium S245 was found to produce high alginate lyase activity. This research was conducted to evaluate activity of the alginate lyase enzyme at various pHs, temperatures and substrates. Polymannuronate and polyguluronate were used to evaluate substrate specificities. Alginate lyase activity was assayed by analysis of reducing sugar released using the 3,5 dinitrosalicylic acid (DNS) method. The research showed that the activity of alginate lyase was optimum at pH of 7.0 and temperature of 45 0C. This enzyme was active for both polymannuronate and polyguluronate susbtrates. The Vmax and Km of this enzyme for polymannuronate and polyguluronate were 200 unit/ml/min and 79.8 mg/ml for polymannuronate substrate and 27.78 unit/ml/min and 13.17 mg/ml for polyguluronate substrate. This enzyme showed unique characteristic in working toward the two substrates.

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Yan, Junjun, Peng Chen, Yan Zeng, Yan Men, Shicheng Mu, Yueming Zhu, Yefu Chen, and Yuanxia Sun. "The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from Marinimicrobium sp. H1." Marine Drugs 17, no. 10 (September 23, 2019): 545. http://dx.doi.org/10.3390/md17100545.

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Alginase lyase is an important enzyme for the preparation of alginate oligosaccharides (AOS), that possess special biological activities and is widely used in various fields, such as medicine, food, and chemical industry. In this study, a novel bifunctional alginate lyase (AlgH) belonging to the PL7 family was screened and characterized. The AlgH exhibited the highest activity at 45 °C and pH 10.0, and was an alkaline enzyme that was stable at pH 6.0–10.0. The enzyme showed no significant dependence on metal ions, and exhibited unchanged activity at high concentration of NaCl. To determine the function of non-catalytic domains in the multi-domain enzyme, the recombinant AlgH-I containing only the catalysis domain and AlgH-II containing the catalysis domain and the carbohydrate binding module (CBM) domain were constructed and characterized. The results showed that the activity and thermostability of the reconstructed enzymes were significantly improved by deletion of the F5/8 type C domain. On the other hand, the substrate specificity and the mode of action of the reconstructed enzymes showed no change. Alginate could be completely degraded by the full-length and modified enzymes, and the main end-products were alginate disaccharide, trisaccharide, and tetrasaccharide. Due to the thermo and pH-stability, salt-tolerance, and bifunctionality, the modified alginate lyase was a robust enzyme which could be applied in industrial production of AOS.

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Cao, Thi Thuy Hang, Mai Nhu Hieu Vo, Duc Thinh Pham, Ngoc Linh Nguyen, Dinh Thuat Nguyen, and Thi Thanh Van Tran. "Study on culture conditionsfor extracellular alginate lyase production by Bacillus velezensis AlgSm1." Ministry of Science and Technology, Vietnam 63, no. 3 (March 30, 2021): 19–23. http://dx.doi.org/10.31276/vjst.63(3).19-23.

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Alginate lyase is an enzyme that degrades alginate to create bioactive oligoalginate for application in medicine, industry, agriculture... In this study, cultural conditions of marine bacteria, Bacillus velezensis AlgSm1 have been investigated for producing alginate lyase with high content and activity. The nutritional components such as carbon source, nitrogen source, initial pH value, and cultural time were investigated. The results showed that the cultural conditions of B. velezensis AlgSm1 to biosynthesize alginate lyase with high activity were 5 mg/ml alginate, 0.8 mg/ml yeast extract; pH 5.5 and 18-hours fermentation at 28-30°C.

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Albrecht, Mark T., and Neal L. Schiller. "Alginate Lyase (AlgL) Activity Is Required for Alginate Biosynthesis in Pseudomonas aeruginosa." Journal of Bacteriology 187, no. 11 (June 1, 2005): 3869–72. http://dx.doi.org/10.1128/jb.187.11.3869-3872.2005.

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ABSTRACT To determine whether AlgL's lyase activity is required for alginate production in Pseudomonas aeruginosa, an algLΔ::Gmr mutant (FRD-MA7) was created. algL complementation of FRD-MA7 restored alginate production, but algL constructs containing mutations inactivating lyase activity did not, demonstrating that the enzymatic activity of AlgL is required for alginate production.

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Miyake, Osamu, Akihito Ochiai, Wataru Hashimoto, and Kousaku Murata. "Origin and Diversity of Alginate Lyases of Families PL-5 and -7 in Sphingomonas sp. Strain A1." Journal of Bacteriology 186, no. 9 (May 1, 2004): 2891–96. http://dx.doi.org/10.1128/jb.186.9.2891-2896.2004.

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ABSTRACT Sphingomonas sp. strain A1 has three endotype alginate lyases (A1-I, A1-II [family PL-7], and A1-III [family PL-5]), each of which is encoded by a single gene. In addition to those of these lyases, a gene (the A1-II′ gene) showing significant identity with the A1-II gene was present in the bacterial genome and coded for an alginate lyase with broad substrate specificity. Since no expression of A1-II′ was observed even in bacterial cells grown on alginate, the A1-II′ gene was thought to be a silent gene derived from the A1-II gene, presumably through duplication, modification, and translocation.

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Zhang, Wen, X. Xia, and Z. Zhang. "Alginate Lyase of a Novel Algae Fermentation Strain." Chemical & biochemical engineering quarterly 33, no. 1 (2019): 125–31. http://dx.doi.org/10.15255/cabeq.2018.1291.

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A novel algae fermentation strain was obtained in our previous work. This strain can produce alginate lyase and alcohol dehydrogenase used for the ethanol fermentation from algae. This research investigated the fermentation, separation and purification of alginate lyase, and the molecular weight of alginate lyase was determined. The optimum conditions for enzyme fermentation were as follows: fermentation medium with 20 g L–1 alginate, initial pH 6.0, and temperature 35 °C. The flasks were cultured in a shaking incubator at 120 rpm for 96 h. The enzyme was purified using the method of salting out, dialysis, and gel chromatography. After purification, the SDS-PAGE method was used to determine the molecular weight of the protein. The molecular weight of alginate lyase was 30–35 KDa. This research contributes to algae biodegradation and fuels production from algae.

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Yang, Suxiao, Zhemin Liu, Xiaodan Fu, Changliang Zhu, Qing Kong, Min Yang, and Haijin Mou. "Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in Pichia pastoris." Marine Drugs 18, no. 6 (June 11, 2020): 305. http://dx.doi.org/10.3390/md18060305.

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Alginate is one of the most abundant polysaccharides in algae. Alginate lyase degrades alginate through a β-elimination mechanism to produce alginate oligosaccharides with special bioactivities. Improving enzyme activity and thermal stability can promote the application of alginate lyase in the industrial preparation of alginate oligosaccharides. In this study, the recombinant alginate lyase cAlyM and its thermostable mutant 102C300C were expressed and characterized in Pichia pastoris. The specific activities of cAlyM and 102C300C were 277.1 U/mg and 249.6 U/mg, respectively. Both enzymes showed maximal activity at 50 °C and pH 8.0 and polyG preference. The half-life values of 102C300C at 45 °C and 50 °C were 2.6 times and 11.7 times the values of cAlyM, respectively. The degradation products of 102C300C with a lower degree of polymerization contained more guluronate. The oligosaccharides with a polymerization degree of 2–4 were the final hydrolytic products. Therefore, 102C300C is potentially valuable in the production of alginate oligosaccharides with specific M/G ratio and molecular weights.

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Yang, Min, Su-Xiao Yang, Zhe-Min Liu, Nan-Nan Li, Li Li, and Hai-Jin Mou. "Rational Design of Alginate Lyase from Microbulbifer sp. Q7 to Improve Thermal Stability." Marine Drugs 17, no. 6 (June 25, 2019): 378. http://dx.doi.org/10.3390/md17060378.

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Alginate lyase degrades alginate by the β-elimination mechanism to produce oligosaccharides with special bioactivities. The low thermal stability of alginate lyase limits its industrial application. In this study, introducing the disulfide bonds while using the rational design methodology enhanced the thermal stability of alginate lyase cAlyM from Microbulbifer sp. Q7. Enzyme catalytic sites, secondary structure, spatial configuration, and molecular dynamic simulation were comprehensively analyzed. When compared with cAlyM, the mutants D102C-A300C and G103C-T113C showed an increase by 2.25 and 1.16 h, respectively, in half-life time at 45 °C, in addition to increases by 1.7 °C and 0.4 °C in the melting temperature, respectively. The enzyme-specific activity and kcat/Km values of D102C-A300C were 1.8- and 1.5-times higher than those of cAlyM, respectively. The rational design strategy that was used in this study provides a valuable method for improving the thermal stability of the alginate lyase.

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Belik, Alexey, Artem Silchenko, Olesya Malyarenko, Anton Rasin, Marina Kiseleva, Mikhail Kusaykin, and Svetlana Ermakova. "Two New Alginate Lyases of PL7 and PL6 Families from Polysaccharide-Degrading Bacterium Formosa algae KMM 3553T: Structure, Properties, and Products Analysis." Marine Drugs 18, no. 2 (February 24, 2020): 130. http://dx.doi.org/10.3390/md18020130.

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A bifunctional alginate lyase (ALFA3) and mannuronate-specific alginate lyase (ALFA4) genes were found in the genome of polysaccharide-degrading marine bacterium Formosa algae KMM 3553T. They were classified to PL7 and PL6 polysaccharide lyases families and expressed in E. coli. The recombinant ALFA3 appeared to be active both on mannuronate- and guluronate-enriched alginates, as well as pure sodium mannuronate. For all substrates, optimum conditions were pH 6.0 and 35 °C; Km was 0.12 ± 0.01 mg/mL, and half-inactivation time was 30 min at 42 °C. Recombinant ALFA4 was active predominately on pure sodium mannuronate, with optimum pH 8.0 and temperature 30 °C, Km was 3.01 ± 0.05 mg/mL. It was stable up to 30 °C; half-inactivation time was 1 h 40 min at 37 °C. 1H NMR analysis showed that ALFA3 degraded mannuronate and mannuronate-guluronate blocks, while ALFA4 degraded only mannuronate blocks, producing mainly disaccharides. Products of digestion of pure sodium mannuronate by ALFA3 at 200 µg/mL inhibited anchorage-independent colony formation of human melanoma cells SK-MEL-5, SK-MEL-28, and RPMI-7951 up to 17% stronger compared to native polymannuronate. This fact supports previous data and suggests that mannuronate oligosaccharides may be useful for synergic tumor therapy.

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Wang, Zhi-Peng, Min Cao, Bing Li, Xiao-Feng Ji, Xin-Yue Zhang, Yue-Qi Zhang, and Hai-Ying Wang. "Cloning, Secretory Expression and Characterization of a Unique pH-Stable and Cold-Adapted Alginate Lyase." Marine Drugs 18, no. 4 (April 1, 2020): 189. http://dx.doi.org/10.3390/md18040189.

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Cold-adapted alginate lyases have unique advantages for alginate oligosaccharide (AOS) preparation and brown seaweed processing. Robust and cold-adapted alginate lyases are urgently needed for industrial applications. In this study, a cold-adapted alginate lyase-producing strain Vibrio sp. W2 was screened. Then, the gene ALYW201 was cloned from Vibrio sp. W2 and expressed in a food-grade host, Yarrowia lipolytica. The secreted Alyw201 showed the activity of 64.2 U/mL, with a molecular weight of approximate 38.0 kDa, and a specific activity of 876.4 U/mg. Alyw201 performed the highest activity at 30 °C, and more than 80% activity at 25–40 °C. Furthermore, more than 70% of the activity was obtained in a broad pH range of 5.0–10.0. Alyw201 was also NaCl-independent and salt-tolerant. The degraded product was that of the oligosaccharides of DP (Degree of polymerization) 2–6. Due to its robustness and its unique pH-stable property, Alyw201 can be an efficient tool for industrial production.

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Yonemoto, Yoshimasa, Kousaku Murata, Akira Kimura, Hisako Yamaguchi, and Kenichi Okayama. "Bacterial alginate lyase: Characterization of alginate lyase-producing bacteria and purification of the enzyme." Journal of Fermentation and Bioengineering 72, no. 3 (January 1991): 152–57. http://dx.doi.org/10.1016/0922-338x(91)90208-x.

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40

Qin, Hui-Min, Takuya Miyakawa, Akira Inoue, Ryuji Nishiyama, Akira Nakamura, Atsuko Asano, Takao Ojima, and Masaru Tanokura. "Structural basis for controlling the enzymatic properties of polymannuronate preferred alginate lyase FlAlyA from the PL-7 family." Chemical Communications 54, no. 5 (2018): 555–58. http://dx.doi.org/10.1039/c7cc06523j.

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Zhou, Hai-Xiang, Shan-Shan Xu, Xue-Jing Yin, Feng-Long Wang, and Yang Li. "Characterization of a New Bifunctional and Cold-Adapted Polysaccharide Lyase (PL) Family 7 Alginate Lyase from Flavobacterium sp." Marine Drugs 18, no. 8 (July 26, 2020): 388. http://dx.doi.org/10.3390/md18080388.

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Alginate oligosaccharides produced by enzymatic degradation show versatile physiological functions and biological activities. In this study, a new alginate lyase encoding gene alyS02 from Flavobacterium sp. S02 was recombinantly expressed at a high level in Yarrowia lipolytica, with the highest extracellular activity in the supernatant reaching 36.8 ± 2.1 U/mL. AlyS02 was classified in the polysaccharide lyase (PL) family 7. The optimal reaction temperature and pH of this enzyme were 30 °C and 7.6, respectively, indicating that AlyS02 is a cold-adapted enzyme. Interestingly, AlyS02 contained more than 90% enzyme activity at 25 °C, higher than other cold-adapted enzymes. Moreover, AlyS02 is a bifunctional alginate lyase that degrades both polyG and polyM, producing di- and trisaccharides from alginate. These findings suggest that AlyS02 would be a potent tool for the industrial applications.

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Chernysheva, Nadezhda, Evgeniya Bystritskaya, Galina Likhatskaya, Olga Nedashkovskaya, and Marina Isaeva. "Genome-Wide Analysis of PL7 Alginate Lyases in the Genus Zobellia." Molecules 26, no. 8 (April 20, 2021): 2387. http://dx.doi.org/10.3390/molecules26082387.

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We carried out a detailed investigation of PL7 alginate lyases across the Zobellia genus. The main findings were obtained using the methods of comparative genomics and spatial structure modeling, as well as a phylogenomic approach. Initially, in order to elucidate the alginolytic potential of Zobellia, we calculated the content of polysaccharide lyase (PL) genes in each genome. The genus-specific PLs were PL1, PL6, PL7 (the most abundant), PL14, PL17, and PL40. We revealed that PL7 belongs to subfamilies 3, 5, and 6. They may be involved in local and horizontal gene transfer and gene duplication processes. Most likely, an individual evolution of PL7 genes promotes the genetic variability of the Alginate Utilization System across Zobellia. Apparently, the PL7 alginate lyases may acquire a sub-functionalization due to diversification between in-paralogs.

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Bakkevig, Karianne, Håvard Sletta, Martin Gimmestad, Randi Aune, Helga Ertesvåg, Kristin Degnes, Bjørn Erik Christensen, Trond E. Ellingsen, and Svein Valla. "Role of the Pseudomonas fluorescens Alginate Lyase (AlgL) in Clearing the Periplasm of Alginates Not Exported to the Extracellular Environment." Journal of Bacteriology 187, no. 24 (December 15, 2005): 8375–84. http://dx.doi.org/10.1128/jb.187.24.8375-8384.2005.

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ABSTRACT Alginate is an industrially widely used polysaccharide produced by brown seaweeds and as an exopolysaccharide by bacteria belonging to the genera Pseudomonas and Azotobacter. The polymer is composed of the two sugar monomers mannuronic acid and guluronic acid (G), and in all these bacteria the genes encoding 12 of the proteins essential for synthesis of the polymer are clustered in the genome. Interestingly, 1 of the 12 proteins is an alginate lyase (AlgL), which is able to degrade the polymer down to short oligouronides. The reason why this lyase is associated with the biosynthetic complex is not clear, but in this paper we show that the complete lack of AlgL activity in Pseudomonas fluorescens in the presence of high levels of alginate synthesis is toxic to the cells. This toxicity increased with the level of alginate synthesis. Furthermore, alginate synthesis became reduced in the absence of AlgL, and the polymers contained much less G residues than in the wild-type polymer. To explain these results and other data previously reported in the literature, we propose that the main biological function of AlgL is to degrade alginates that fail to become exported out of the cell and thereby become stranded in the periplasmic space. At high levels of alginate synthesis in the absence of AlgL, such stranded polymers may accumulate in the periplasm to such an extent that the integrity of the cell is lost, leading to the observed toxic effects.

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Yonemoto, Yoshimasa, Hirokazu Tanaka, Tomohiro Hisano, Kenji Sakaguchi, Shiro Abe, Tetsuo Yamashita, Akira Kimura, and Kousaku Murata. "Bacterial alginate lyase gene: Nucleotide sequence and molecular route for generation of alginate lyase species." Journal of Fermentation and Bioengineering 75, no. 5 (January 1993): 336–42. http://dx.doi.org/10.1016/0922-338x(93)90130-z.

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Kim, Se Won, Gun-Do Kim, and Soo-Wan Nam. "Coexpression of Alginate Lyase with Hyperthermophilic Archaea Chaperonin in E. coli." Journal of Life Science 25, no. 2 (February 28, 2015): 130–35. http://dx.doi.org/10.5352/jls.2015.25.2.130.

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Yang, Jin, Dandan Cui, Diwen Chen, Wenkang Chen, Shuo Ma, and Hong Shen. "Purification and Characterization of a Novel Endolytic Alginate Lyase from Microbulbifer sp. SH-1 and Its Agricultural Application." Marine Drugs 18, no. 4 (March 31, 2020): 184. http://dx.doi.org/10.3390/md18040184.

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Alginate, an important acidic polysaccharide in marine multicellular algae, has attracted attention as a promising biomass resource for the production of medical and agricultural chemicals. Alginate lyase is critical for saccharification and utilization of alginate. Discovering appropriate and efficient enzymes for depolymerizing alginate into fermentable fractions plays a vital role in alginate commercial exploitation. Herein, a unique alginate lyase, AlgSH7, belonging to polysaccharide lyase 7 family is purified and characterized from an alginate-utilizing bacterium Microbulbifer sp. SH-1. The purified AlgSH7 shows a specific activity of 12,908.26 U/mg, and its molecular weight is approximately 66.4 kDa. The optimal temperature and pH of AlgSH7 are 40 °C and pH 9.0, respectively. The enzyme exhibits stability at temperatures below 30 °C and within an extensive pH range of 5.0–9.0. Metal ions including Na+, K+, Al3+, and Fe3+ considerably enhance the activity of the enzyme. AlgSH7 displays a preference for poly-mannuronic acid (polyM) and a very low activity towards poly-guluronic acid (polyG). TLC and ESI-MS analysis indicated that the enzymatic hydrolysates mainly include disaccharides, trisaccharides, and tetrasaccharides. Noteworthy, the alginate oligosaccharides (AOS) prepared by AlgSH7 have an eliciting activity against chilling stress in Chinese flowering cabbage (Brassica parachinensis L.). These results suggest that AlgSH7 has a great potential to design an effective process for the production of alginate oligomers for agricultural applications.

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Kim, Geun-Hyub, Sung-Mok Lee, Soo-Jeong Choi, and Jae-Hwa Lee. "Isolation and Characteristics of Alginate lyase Producing Microorganism: Sanguibacter keddieii NC9." KSBB Journal 26, no. 6 (December 31, 2011): 538–42. http://dx.doi.org/10.7841/ksbbj.2011.26.6.538.

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Shibata, Toshiyuki, Reona Fujii, Yoshihiko Nishioka, Hideo Miyake, Tetsushi Mori, and Reiji Tanaka. "A Simple Analysis Method for 4-Deoxy-l-erythro-5-hexoseulose Uronic Acid by HPLC-ELSD with Column for Anion Analysis." Natural Product Communications 14, no. 5 (May 2019): 1934578X1985099. http://dx.doi.org/10.1177/1934578x19850990.

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4-Deoxy-l- erythro-5-hexoseulose uronic acid (DEH) is a rare deoxy sugar produced from alginate by the action of an exotype alginate lyase. A simple and rapid method for analyzing DEH using high-performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) was developed in this study. For chromatography, an isocratic elution of ammonium formate buffer including formic acid and a column for anion chromatography were used. In the developed method, DEH was detected at a retention time of 3.038 minutes and limits of detection (signal-noise ratio = 3) and quantification (signal-noise ratio = 10) were 37.5 and 124.9 µg/mL as a sodium DEH, respectively. In addition, separation and detection of alginate unsaturated oligosaccharides were also tested using the method. Within an analysis time of 10 minutes, it was possible to separate and detect unsaturated disaccharide, unsaturated trisaccharide, and unsaturated tetrasaccharide prepared using poly(β-d-mannuronate) lyase and sodium alginate of high mannuronate type. The HPLC-ELSD method established in this study will be applicable for quantitative analysis of DEH and measurement of exotype alginate lyase activity.

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Li, Shangyong, Yanan Wang, Xiao Li, Beom Suk Lee, Samil Jung, and Myeong-Sok Lee. "Enhancing the Thermo-Stability and Anti-Biofilm Activity of Alginate Lyase by Immobilization on Low Molecular Weight Chitosan Nanoparticles." International Journal of Molecular Sciences 20, no. 18 (September 14, 2019): 4565. http://dx.doi.org/10.3390/ijms20184565.

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Bacterial biofilm causes severe antibiotic resistance. An extracellular polymeric substance (EPS) is the main component in the bacterial biofilm. Alginate is a key EPS component in the biofilm of Pseudomonas aeruginosa and responsible for surface adhesion and stabilization of biofilm. Alginate lyase has emerged as an efficient therapeutic strategy targeting to degrade the alginate in the biofilm of P. aeruginosa. However, the application of this enzyme is limited by its poor stability. In this study, chitosan nanoparticles (CS-NPs) were synthesized using low molecular weight chitosan and alginate lyase Aly08 was immobilized on low molecular weight chitosan nanoparticles (AL-LMW-CS-NPs). As a result, the immobilization significantly enhanced the thermal stability and reusability of Aly08. In addition, compared with free Aly08, the immobilized AL-LMW-CS-NPs exhibited higher efficiency in inhibiting biofilm formation and interrupting the established mature biofilm of P. aeruginosa, which could reduce its biomass and thickness confirmed by confocal microscopy. Moreover, the biofilm disruption greatly increased the antibiotic sensitivity of P. aeruginosa. This research will contribute to the further development of alginate lyase as an anti-biofilm agent.

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Kim, Hee Sook, Choul-Gyun Lee, and Eun Yeol Lee. "Alginate lyase: Structure, property, and application." Biotechnology and Bioprocess Engineering 16, no. 5 (September 27, 2011): 843–51. http://dx.doi.org/10.1007/s12257-011-0352-8.

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Journal articles on the topic 'Alginate lyase'

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