Academic literature on the topic 'Enzyme-assisted'

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Journal articles on the topic "Enzyme-assisted"

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Jacques, Suzanne L., I. Ahmad Mirza, Linda Ejim, et al. "Enzyme-Assisted Suicide." Chemistry & Biology 10, no. 10 (2003): 989–95. http://dx.doi.org/10.1016/j.chembiol.2003.09.015.

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Riemenschneider, Leif, Sven Blank, and Manfred Radmacher. "Enzyme-Assisted Nanolithography." Nano Letters 5, no. 9 (2005): 1643–46. http://dx.doi.org/10.1021/nl0484550.

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Soremekun, Olanrewaju A., Melissa L. Shear, Sagar Patel, et al. "Rapid vascular glucose uptake via enzyme-assisted subcutaneous infusion: Enzyme-Assisted Subcutaneous Infusion Access Study." American Journal of Emergency Medicine 27, no. 9 (2009): 1072–80. http://dx.doi.org/10.1016/j.ajem.2008.08.028.

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Nielsen, Per Munk. "Phospholipase for enzyme-assisted alkaline refining." INFORM International News on Fats, Oils, and Related Materials 32, no. 1 (2021): 22–24. http://dx.doi.org/10.21748/inform.01.2021.22.

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MOORHEAD, LOUISE C., and NORMAN RADTKE. "ENZYME-ASSISTED VITRECTOMY WITH BACTERIAL COLLAGENASE." Retina 5, no. 2 (1985): 98–100. http://dx.doi.org/10.1097/00006982-198500520-00007.

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Dahmer, Eric R. "Microcomputer assisted teaching of enzyme kinetics." Biochemical Education 15, no. 2 (1987): 69–70. http://dx.doi.org/10.1016/0307-4412(87)90087-2.

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Fronza, Giovanni, Claudio Fuganti, Piero Grasselli, Giuseppe Pedrocchi-Fantoni, and Stefano Servi. "Enzyme assisted synthesis of (S)-sotolon." Tetrahedron Letters 33, no. 38 (1992): 5625–28. http://dx.doi.org/10.1016/s0040-4039(00)61164-4.

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Qadir, Rahman, Farooq Anwar, Mazhar Amjad Gilani, Sadaf Zahoor, Muhammad Misbah ur Rehman, and Muhammad Mustaqeem. "RSM/ANN based optimized recovery of phenolics from mulberry leaves by enzyme-assisted extraction." Czech Journal of Food Sciences 37, No. 2 (2019): 99–105. http://dx.doi.org/10.17221/147/2018-cjfs.

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Recovery of phenolics from Morus alba leaves (MAL) and extraction into the solvent was optimized using enzyme-assisted extraction. The influence of four parameters, including enzyme concentration (EC), temperature (T), incubation time (t) and pH were investigated using rotatable central composite design (RCCD). Two factors, namely enzyme concentration and pH, exhibited significant effect on extraction efficacy yield of extractable phenolics from MAL. Furthermore, artificial neural network (ANN) model was executed to predict the relationship between dependent and independent variables. Among enzyme complexes (kemzyme dry-plus, natuzyme and zympex-014) employed for extraction, zympex-014 assisted extract depicted maximum amount of phenolic bioactives from MAL. Morphological changes in the cell wall of MAL residue were elucidated by scanning electron microscopy (SEM). The main phenolic compounds identified and quantified by gas chromatography mass spectrometry (GC/MS) in MAL extract were found to be quercetin, gallic acid, m-coumaric acid, cinnamic acid, syrinigc acid and vanillic acid.
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Singh, Gurpreet, Mohit Kumar, Ruchika Zalpouri, Pratik Pandit Potdar, Kamalpreet Singh, and Kulwinder Kaur. "Effects of different aqueous extraction techniques on physicochemical quality and oil recovery of sesame oil." Environment Conservation Journal 24, no. 1 (2023): 136–42. http://dx.doi.org/10.36953/ecj.11892309.

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Sesame is the oldest oilseed crop in agriculture, and it produces more oil than any other crop on the planet. This research aimed to investigate the impact of different oil extraction procedures on sesame seed oil physicochemical quality and oil recovery. The oil was extracted from the clean and healthy seeds using four extraction methods: aqueous, enzyme-assisted aqueous, ultrasound-assisted aqueous and solvent extraction using the Soxhlet apparatus. It was observed that ultrasound-assisted aqueous extracted oil had maximum saponification value and minimum acid value, refractive index, and lower free fatty acid value, compared with aqueous extracted oil and enzyme-assisted aqueous extracted oil. Ultrasound-assisted aqueous extraction method also yielded maximum oil, retrieval followed by enzyme-assisted aqueous extraction and aqueous extraction.
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Wirajana, I. N., N. M. T. Juliasari, A. A. I. A. M. Laksmiwati, and N. W. Bogoriani. "SUHU DAN WAKTU OPTIMUM PROSES EKSTRAKSI ANTOSIANIN DALAM UBI JALAR UNGU (Ipomoea batatas L.) DENGAN ?-L-ARABINOFURANOSIDASE." Jurnal Kimia 13, no. 1 (2019): 88. http://dx.doi.org/10.24843/jchem.2019.v13.i01.p14.

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Enzyme-assisted extraction (EAE) method is one of the most environmentally friendly methods of enzyme application in the extraction of bioactive compounds. The purpose of this study was to determine the optimum temperature and time required in the extraction of anthocyanin compounds from purple sweet potato (Ipomoea batatas L.) with and without ?-L-arabinofuranosidase (AbfA) - assisted. The AbfA enzyme was obtained from Saccharomyces cerevisiae recombinant strain BJ1824 contain pYHMI-Af plasmid. The optimum temperature and time in the extraction of anthocyanin compound with and without ?-L-arabinofuranosidase from purple sweet potato were performed on the 40, 50, 60 and 700C; and 150, 180, 210 minutes. The extraction was done by ethanol solvent of 60,32% (v/v) acidified with citric acid of 2,39% (b/v). The measurement of anthocyanin levels using UV-Vis Spectrophotometer at 527 nm and 700 nm wavelengths at pH 1,0 and 4,5. The optimum condition of non-enzyme-assisted extraction was at 600C for 210 minutes, with the anthocyanin levels of 26,3842 mg/L; while with the AbfA enzyme-assisted at 500C for 180 minutes, with the anthocyanin levels of 28,2056 mg/L. The extraction with enzyme-assisted resulted the anthocyanin levels of 6,90% higher than without the using of enzyme.
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Dissertations / Theses on the topic "Enzyme-assisted"

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Williams, Richard James. "Enzyme assisted self-assembly." Thesis, University of Manchester, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.496231.

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Self-assembling peptide systems provide a pathway for the formation of complex molecular assemblies from relatively simple designed molecules. Stimuli which have been used to trigger the self-assembly (SA) process in aqueous conditions include temperature, pH, ionic strength, and solvent exchange. Additionally, enzymes may be used to selectively control the self-assembly process; enzymes are uniquely chemo-, regio-, and enantioselective, and work naturally under mild conditions without disrupting biological Interactions.
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Hughes, Meghan. "Peptide nanomaterials : characterisation of enzyme-assisted self-assembly." Thesis, University of Strathclyde, 2012. http://oleg.lib.strath.ac.uk:80/R/?func=dbin-jump-full&object_id=18209.

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tan, yi. "Enzyme Catalyzed and Ultrasound Assisted Transformation of Selected Pollutants." FIU Digital Commons, 2017. http://digitalcommons.fiu.edu/etd/3220.

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The widespread use of synthetic drugs and as feed additives has resulted in the release of large amounts of biologically active chemicals into the environment. Exposure to environmentally relevant concentrations of chemicals can have severe effects on human health. Therefore, effective degradation of these synthetic, biologically active compounds is of paramount importance. Diphenhydramine (DPH) has been selected as a target compound for ultrasound remediation. The results demonstrated that ultrasound-induced degradation has potential applications in managing aqueous media contaminated with DPH. Atorvastatin and roxarsone have been selected as representative substrates for chloroperoxidase (CPO) catalyzed transformation of pollutants. These studies demonstrate atorvastatin and roxarsone can be degraded efficiently by CPO. The transformation products of each compound were identified and the mechanisms of CPO catalysis postulated. This study provides a foundation for assessing the feasibility of applying CPO in the remediation of water and soil contaminated with pharmaceuticals and feed additives.
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Kuuranne, Tiia. "Anabolic steroid glucuronides : enzyme-assisted synthesis and liquid chromatographic-mass spectrometric analysis." Helsinki : University of Helsinki, 2003. http://ethesis.helsinki.fi/julkaisut/mat/farma/vk/kuuranne/.

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Campbell, Kerry Alan. "Protein and oil recoveries from enzyme-assisted aqueous extraction of soybeans and sunflower seed." [Ames, Iowa : Iowa State University], 2010. http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqdiss&rft_dat=xri:pqdiss:3403074.

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Kapasakalidis, Petros G. "Enzyme assisted extraction of antioxidant phenols and anthocyanins from blackcurrant (Ribes nigrum L.) press residues." Thesis, University of Reading, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.433443.

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Kalathoor, Roschni [Verfasser]. "Quantitative and Enantioselective Analyses Following Enzyme Assisted Processes in Soil: Degradation and Non-extractable Residue Formation of the Fungicide Metalaxyl / Roschni Kalathoor." München : Verlag Dr. Hut, 2016. http://d-nb.info/1103872990/34.

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Li, Yuxiu. "Technology of enzyme extraction of Gynostemma pentaphyllum saponins and studies on its hypoglycemic activity." Магістерська робота, Kyiv National University of Technology and Design, 2021. https://er.knutd.edu.ua/handle/123456789/19271.

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In this master's thesis, the composition of saponins in extracts of Gynostemma pentaphyllum obtained by enzymatic extraction has been determined. The HPLC method was used for the simultaneous analysis of hypenosides in medicinal plant materials of different periods and places of growth. The results showed that the highest content of hypenosides was noted in August, the lowest in May; the composition and content of hypenosides from different production zones in the same growing season were different. The optimal parameters of extraction using hypenosidase are as follows: the time of enzymatic hydrolysis is 100 minutes, the temperature of enzymatic hydrolysis is 45℃, the concentration of pectinase is 2 mg/ml. During the extraction process, pectinase transformed the saponins of gynostemma pentaphyllum, making them easier to isolate from plant cells. It has been proven that gynostemma saponins inhibit the activity of alpha-glycosidase with an inhibition rate of more than 90% compared to acarbose and have the effect of lowering blood sugar levels.<br>У цій роботі визначено склад сапонінів в екстрактах Gynostemma pentaphyllum, отриманих шляхом ферментативної екстракції. Метод ВЕРХ використано для одночасного аналізу гіпенозидів у лікарській рослинній сировині різних періодів і місць зростання. Результати показали, що найвищий вміст гіпенозидів відмічений у серпні, найменший у травні; склад і вміст гіпенозидів з різних зон виробництва в одному вегетаційному періоді були різними. Оптимальні параметри екстракції за допомогою гіпенозидази такі: час ферментативного гідролізу 100 хвилин, температура ферментативного гідролізу 45℃, концентрація пектинази 2 мг/мл. Під час процесу екстракції пектиназа трансформувала сапоніни Gynostemma pentaphyllum, завдяки чому їх легше було виділити з рослинних клітин. Доведено, що сапоніни гіностеми пригнічують активність альфа-глікозидази зі швидкістю інгібування більше ніж на 90% порівняно з акарбозою та впливають на зниження рівня цукру в крові.
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Liang, Pingping. "Gold Nanoparticle-Based Colorimetric Sensors for Detection of DNA and Small Molecules." FIU Digital Commons, 2016. http://digitalcommons.fiu.edu/etd/2595.

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Biosensors have proven to be a powerful tool for detecting diverse targets, such as proteins, DNA, and small molecules representing disease biomarkers, toxins, drugs and their metabolites, environmental pollutants, agrichemicals, and antibiotics with high sensitivity and specificity. The major objective of the research described in this dissertation was to develop low cost, low sample volume, highly sensitive and specific AuNP-based colorimetric sensor platforms for the detection of DNA and small molecules. With this in mind, we propose an instrument-free approach in chapter three for the detection of NADH with a sensor constructed on a paper substrate, based on the target-induced inhibition of AuNP dissolution. The successful detection of this important molecule opens the door to numerous possibilities for dehydrogenase characterization, because NAD+/NADH are essential cofactors for more than 300 dehydrogenase enzymes. To further increase the sensitivity of our hybridization-based assay for DNA detection, we developed an enzyme-assisted target recycling (EATR) strategy in chapter four and have applied such an EATR-based colorimetric assay to detect single-nucleotide mismatches in a target DNA with DNA-functionalized AuNPs. This assay is based on the principle that nuclease enzymes recognize probe–target complexes, cleaving only the probe strand. This results in target release, enabling subsequent binding to and cleavage of another probe molecule. When the probe is conjugated onto AuNPs, complete cleavage from the AuNP surface produces a detectable signal in high ionic strength environments as the nanoparticles undergo aggregation. With such enzyme-assisted amplification, target detection can occur with a very low nM detection limit within 15 minutes. The extent of DNA loading on the AuNP surface plays an important role in the efficiency of DNA hybridization and aptamer-target assembly. Many studies have shown that high surface-coverage is associated with steric hindrance, electrostatic repulsive interactions and elevated surface salt concentration, whereas low surface-coverage can result in nonspecific binding of oligonucleotides to the particle surface. In chapter five, we investigated DNA surface coverage effects, and apply this optimization in conjunction with a highly-specific aptamer to develop a sensitive colorimetric sensor for rapid cocaine detection based on the inhibition of nuclease enzyme activity.
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Fournière, Mathilde. "Extraction et évaluation des propriétés biologiques des ulvanes et oligo-ulvanes de l'algue verte Ulva sp. : actions sur le métabolisme de cellules cutanées et sur le microbiote cutané." Electronic Thesis or Diss., Lorient, 2021. http://www.theses.fr/2021LORIS593.

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Les ulvanes sont des polysaccharides matriciels sulfatés constitutifs de la paroi de l’algue verte Ulva sp.. Bien que proliférative et quantitativement disponible, Ulva sp. est peu valorisée avec des applications dans des domaines à faible valeur ajoutée. Ce projet de thèse a pour objectif la production de fractions enrichies en ulvanes et en oligo-ulvanes dans le but d’évaluer leurs propriétés biologiques sur le système cutané. Les fractions d’ulvanes ont été produites par macération et extraction assistée par enzyme suivie de différents procédés de purification : précipitation éthanolique et dialyse. Des dépolymérisations radicalaire et acide ont permis d’obtenir des fractions d’oligo-ulvanes. Ces fractions d’ulvanes et d’oligo-ulvanes stimulent la prolifération de fibroblastes dermiques humains. Une augmentation de la synthèse protéique de composants matriciels dont les collagènes de type I et III, et les glycosaminoglycanes a été mise en évidence. Les fractions stimulent également l’expression, la synthèse et l’activité enzymatique de MMP-1. Au niveau du microbiote cutané, ces fractions n’altèrent pas la croissance des bactéries commensales S. epidermidis, S. aureus et C. acnes, mais peuvent modifier la formation de biofilm. Les fractions d’ulvanes et d’oligo-ulvanes diminuent également le potentiel inflammatoire des kératinocytes HaCaT induit par C. acnes. Ainsi, les travaux ont démontré que les fractions d’ulvanes et d’oligo-ulvanes présentent des activités biologiques prometteuses pour des applications dermo-cosmétiques dans le cadre de stratégies de réparation tissulaire, anti-vieillissement, ou anti-inflammatoire, dans le respect du microbiote commensal cutané<br>Ulvans are matrix sulfated polysaccharides constitutive of green seaweed Ulva sp. cell wall. Although proliferative and quantitatively available, Ulva sp. is little valued with applications in low added value sectors. The objective of this project is to produce fractions enriched in ulvans and oligo-ulvans in order to evaluate their biological properties on cutaneous system. Ulvan fractions were produced by maceration and enzyme-assisted extraction followed by different purification processes: ethanolic precipitation and dialysis. Radical and acid depolymerization processes allowed to obtain oligo-ulvan fractions. Ulvan and oligo-ulvan fractions have been shown to stimulate human dermal fibroblasts proliferation. An increase in the synthesis of extracellular matrix components such as type I and III collagens, and glycosaminoglycans has been demonstrated. Fractions also stimulate the expression, synthesis and MMP-1 enzymatic activity. At the skin microbiota level, the fractions do not alter the growth of commensal bacteria S. epidermidis, S. aureus and C. acnes, but may modify the biofilm formation. Ulvan and oligo-ulvan fractions also reduce the inflammatory potential of HaCaT keratinocytes induced by C. acnes. Thus, this work has shown that the ulvan and oligo-ulvan fractions exhibit promising biological activities for dermo-cosmetic applications in tissue repair, anti-ageing or anti-inflammatory strategies while preserving commensal skin microbiota
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Books on the topic "Enzyme-assisted"

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Enzyme Assisted Organic Synthesis. CRC, 2002.

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Book chapters on the topic "Enzyme-assisted"

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Maled, Sadhana B., Ajay R. Bhat, Subrahmanya Hegde, Yuvaraj Sivamani, Arunachalam Muthuraman, and Sumitha Elayaperumal. "Enzyme-Assisted Extraction." In Bioactive Extraction and Application in Food and Nutraceutical Industries. Springer US, 2024. http://dx.doi.org/10.1007/978-1-0716-3601-5_8.

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Simpson, Benjamin K., Xin Rui, and Jiang XiuJie. "Enzyme-assisted food processing." In Food Engineering Series. Springer US, 2011. http://dx.doi.org/10.1007/978-1-4614-1587-9_13.

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Radtke, Norman D., Michael T. Tseng, Kang-Nian Liu, and Louise C. Moorhead. "Enzyme-Assisted Vitrectomy: An Update." In Proliferative Vitreoretinopathy (PVR). Springer New York, 1988. http://dx.doi.org/10.1007/978-1-4612-3910-9_20.

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Marathe, Sandesh J., Swati B. Jadhav, Sandip B. Bankar, and Rekha S. Singhal. "Enzyme-Assisted Extraction of Bioactives." In Food Bioactives. Springer International Publishing, 2017. http://dx.doi.org/10.1007/978-3-319-51639-4_8.

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Jimat, Dzun Noraini, Nur Huda Syazwani Jafri, Wan Mohd Fazli Wan Nawawi, and Yusilawati Ahmad Nor. "Enzyme-Assisted Cellulose Nanofibers Production." In Handbook of Biorefinery Research and Technology: Biomass Logistics to Saccharification. Springer Netherlands, 2024. http://dx.doi.org/10.1007/978-94-007-6308-1_82.

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Jimat, Dzun Noraini, Nur Huda Syazwani Jafri, Wan Mohd Fazli Wan Nawawi, and Yusilawati Ahmad Nor. "Enzyme-Assisted Cellulose Nanofibers Production." In Handbook of Biorefinery Research and Technology. Springer Netherlands, 2024. http://dx.doi.org/10.1007/978-94-007-6724-9_82-1.

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Zhou, Yong-Kang, and Ji-Shen Zheng. "Enzyme-Assisted Depolymerization of Polymers." In ACS Symposium Series. American Chemical Society, 2025. https://doi.org/10.1021/bk-2025-1501.ch005.

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Andersch, P., F. Fazio, B. Haase, B. Jakob, and M. P. Schneider. "Enzyme Assisted Routes to Bioactive Molecules." In Enzymes in Action. Springer Netherlands, 2000. http://dx.doi.org/10.1007/978-94-010-0924-9_2.

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Kleekayai, Thanyaporn, Mohammadreza Khalesi, Miryam Amigo-Benavent, Maria Cermeño, Pádraigín Harnedy-Rothwell, and Richard J. FitzGerald. "Enzyme-Assisted Extraction of Plant Proteins." In Green Protein Processing Technologies from Plants. Springer International Publishing, 2023. http://dx.doi.org/10.1007/978-3-031-16968-7_6.

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Berger, M., B. Jakob, C. Waldinger, and M. P. Schneider. "Enzyme Assisted Synthesis of Delivery Systems." In Targeting of Drugs 5. Springer US, 1996. http://dx.doi.org/10.1007/978-1-4615-6405-8_19.

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Conference papers on the topic "Enzyme-assisted"

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Kim, Youngmi, and Virginia Lee. "Enzyme-assisted Protein Extraction from Alfalfa Leaves." In 2021 ASABE Annual International Virtual Meeting, July 12-16, 2021. American Society of Agricultural and Biological Engineers, 2021. http://dx.doi.org/10.13031/aim.202100024.

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Cheng, Xian, Liangwu Bi, Zhendong Zhao, and Yuxiang Chen. "Advances in Enzyme Assisted Extraction of Natural Products." In 3rd International Conference on Material, Mechanical and Manufacturing Engineering (IC3ME 2015). Atlantis Press, 2015. http://dx.doi.org/10.2991/ic3me-15.2015.72.

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Greffe, Lionel, and Hugues Driguez. "ENZYME-ASSISTED SYNTHESES OF NOVEL AMYLOPECTIN AND LIMIT DEXTRIN ANALOGUES." In XXIst International Carbohydrate Symposium 2002. TheScientificWorld Ltd, 2002. http://dx.doi.org/10.1100/tsw.2002.408.

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Gutierrez, Diana, Antonio Cruz, Xueli Chen, et al. "Enzyme-assisted liquefaction for different fractions of corn stover pellets." In American Chemical Society Spring Meeting 2023-Crossroads of Chemistry / Indianapolis, IN / 3-29-23. US DOE, 2023. https://doi.org/10.2172/2283677.

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Cheng, G., S. J. Hao, and S. Y. Zheng. "Construction of graphene-based enzyme system for rapid photo-assisted proteolysis." In TRANSDUCERS 2015 - 2015 18th International Solid-State Sensors, Actuators and Microsystems Conference. IEEE, 2015. http://dx.doi.org/10.1109/transducers.2015.7181188.

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Alotaibi, Emran, Mohamed G. Arab, Marwan Naeem, Maher Omar, and Omar Mostafa. "Nano-Assisted Enzyme-Induced Calcium Precipitation (EICP) for Acidic Soil Improvement." In Geo-Congress 2024. American Society of Civil Engineers, 2024. http://dx.doi.org/10.1061/9780784485330.002.

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Gutierrez, Diana, Antonio Cruz, Xueli Chen, et al. "Liquefaction of different corn stover fractions assisted by enzyme-biomass deconstruction." In 45th Symposium on Biomaterials, Fuels and Chemicals / Portland, OR / 4-30-2023. US DOE, 2023. https://doi.org/10.2172/2283718.

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"Techno-Economic Analysis of Integrated Enzyme Assisted Aqueous Extraction of Soybean Oil." In 2016 ASABE International Meeting. American Society of Agricultural and Biological Engineers, 2016. http://dx.doi.org/10.13031/aim.20162459761.

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Sarasini, F., J. Tirillò, G. Maffei, A. Zuorro, R. Lavecchia, and M. Abdelhak. "Enzyme-assisted extraction of fibres from Ampelodesmos mauritanicus and mechanical characterization of their composites." In 9TH INTERNATIONAL CONFERENCE ON “TIMES OF POLYMERS AND COMPOSITES”: From Aerospace to Nanotechnology. Author(s), 2018. http://dx.doi.org/10.1063/1.5045882.

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Iltchenko, Nikita, Jesse Beam, and Ying Zha. "Applications and benefits of phospholipase A enzymes in seed oil processing." In 2022 AOCS Annual Meeting & Expo. American Oil Chemists' Society (AOCS), 2022. http://dx.doi.org/10.21748/rrjs3474.

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Current market conditions have further driven focus on efficiency for oilseed producers. Phospholipases use for enzymatic degumming and refining has, therefore, become more attractive than ever. Since its introduction a decade ago, enzyme assisted seed oil processing has been demonstrated at many plants around the globe for its benefit on yield increase. With the learnings gained from the field, enzyme producers have brought out new generations of products to improve performance, as well as meeting new requirements of the oil plants, such as lower chemical usage, less byproducts and higher ease of use. We would like to demonstrate the applications and benefits of two new phospholipase A enzymes, being Phospholipase A1 (Purifine® PLA1) and Phospholipase A2 (Purifine® LM), which offers these new benefits to producers, crushing and refining. Often the biggest hurdle encountered in implementing enzyme technology is capital expenditure. DSM has worked to develop options for nearly all plants to ensure benefits from enzymatic degumming can be appreciated across the industry. The applications of these enzymes, including efforts needed to make plant changes to accommodate enzyme usage, are demonstrated herein.
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Reports on the topic "Enzyme-assisted"

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Fridman, Eyal, and Eran Pichersky. Tomato Natural Insecticides: Elucidation of the Complex Pathway of Methylketone Biosynthesis. United States Department of Agriculture, 2009. http://dx.doi.org/10.32747/2009.7696543.bard.

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Plant species synthesize a multitude of specialized compounds 10 help ward off pests. and these in turn may well serve as an alternative to synthetic pesticides to reduce environmental damage and health risks to humans. The general goal of this research was to perform a genetic and biochemical dissection of the natural-insecticides methylketone pathway that is specific to the glandular trichomes of the wild species of tomato, Solanumhabrochaites f. glabratum (accession PI126449). Previous study conducted by us have demonstrated that these compounds are synthesized de novo as a derivate pathway of the fatty acid biosynthesis, and that a key enzyme. designated MethylketoneSynthase 1 (MKS 1). catalyzes conversion of the intermediate B-ketoacyl- ACPs to the corresponding Cn-1 methylketones. The approach taken in this proposed project was to use an interspecific F2 population. derived from the cross between the cultivated lV182 and the wild species PIl26449. for three objectives: (i) Analyze the association between allelic status of candidate genes from the fatty acid biosynthesis pathway with the methylketone content in the leaves (ii) Perform bulk segregant analysis of genetic markers along the tomato genome for identifying genomic regions that harbor QTLs for 2TD content (iii) Apply differential gene expression analysis using the isolated glands of bulk segregant for identifying new genes that are involved in the pathway. The genetic mapping in the interspecific F2 population included app. 60 genetic markers, including the candidate genes from the FAS pathway and SSR markers spread evenly across the genome. This initial; screening identified 5 loci associated with MK content including the candidate genes MKS1, ACC and MaCoA:ACP trans. Interesting observation in this genetic analysis was the connection between shape and content of the glands, i.e. the globularity of the four cells, typical to the wild species. was associated with increased MK in the segregating population. In the next step of the research transcriptomic analysis of trichomes from high- and 10w-MK plants was conducted. This analysis identified a new gene, Methy1ketone synthase 2 (MKS2), whose protein product share sequence similarity to the thioesterase super family of hot-dog enzymes. Genetic analysis in the segregating population confirmed its association with MK content, as well as its overexpression in E. coli that led to formation of MK in the media. There are several conclusions drawn from this research project: (i) the genetic control of MK accumulation in the trichomes is composed of biochemical components in the FAS pathway and its vicinity (MKS 1 and MKS2). as well as genetic factors that mediate the morphology of these specialized cells. (ii) the biochemical pathway is now realized different from what was hypothesized before with MKS2 working upstream to I\1KS 1 and serves as the interface between primary (fatty acids) and secondary (MK) metabolism. We are currently testing the possible physical interactions between these two proteins in vitro after the genetic analysis showed clear epistatic interactions. (iii) the regulation of the pathway that lead to specialized metabolism in the wild species is largely mediated by transcription and one of the achievements of this project is that we were able to isolate and verify the specificity of the MKS1 promoter to the trichomes which allows manipulation of the pathways in these cells (currently in progress). The scientific implications of this research project is the advancement in our knowledge of hitherto unknown biochemical pathway in plants and new leads for studying a new family in plants (hot dog thioesterase). The agricultural and biotechnological implication are : (i) generation of new genetic markers that could assist in importing this pathway to cultivated tomato hence enhancing its natural resistance to insecticides, (ii) the discovery of MKS2 adds a new gene for genetic engineering of plants for making new fatty acid derived compounds. This could be assisted with the use of the isolated and verified MKS1 promoter. The results of this research were summarized to a manuscript that was published in Plant Physiology (cover paper). to a chapter in a proceeding book. and one patent was submitted in the US.
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