Academic literature on the topic 'Exozyme'

Indonesia has abundant ethanol biomass feedstocks. However the second-generation ethanol production process is still hampered by the unavailability of cellulase enzyme in the process of decomposition of lignocellulose into saccharides that can be processed into ethanol through fermentation. Cellulase is known as exozyme produced by Bacillus sp. in submerged fermentation. In this study, cellulase production by Bacillus sp. CC BPPT RK2 on natural and abundant agricultural waste substrates (rice bran and coconut water) was evaluated by investigating the optimum conditions for cellulase production in a 50 ml laboratory scale. Preliminary test using Luria Bentani (LB) medium with additional CMC (1%) were done to select optimum range of pH and Temperature. The preliminary tests results were then followed by optimization of pH and temperature, which were carried out using response surface methodology (RSM). RSM optimization model showed optimum values 6.23 for pH and 40.04 °C, with 14 terms (each with 1 degree of freedom), 4 linear effects, 6 interaction effects and 4 quadratic effects. These optimization by RSM results were slightly different compared to preliminary test, showing the effect of interactions between parameters. The characteristics of interaction among variables tested against the cellulase activity are reported in this study including: positive effects on cellulase activity of the resulting responses; negative interactions affecting the response of cellulase activity; synergistic interaction; and antagonistic interactions between each other.

Background and Aim: In Nepal not much of study of Thermophilic area and Thermophiles have been done. Thermophilic bacteria are less studied but are important group of microorganisms due to their ability to produce industrially important enzymes. Methods: In this study, thermophilic bacteria were isolated from hot spring of Bhurung, Nepal. Wide range of bacteria that could grow at high temperatures and tolerate extreme temperature were characterized by morphology, biochemistry and sequencing of its 16S rRNA gene sequence. The isolates were screened for production of extracellular enzymes like protease, amylase, lipase, cellulase, caseinase, pectinase and xylanase activity. Phylogenetic tree construction and G+C content evaluation of the isolate was also studied.Results: 15 isolates with ability to tolerate high temperatures were identified as Bacillus sp. by morphology, biochemistry and sequencing of its 16S rRNA gene sequence. BLAST search analysis of the sequence was performed and result showed maximum identity (99% similarity) with Bacillus licheniformis, Bacillus subtilis and Bacillus pumilus. Isolated strains exhibited considerable amount of extracellular exozymes activity. Phylogenetic analysis of the isolates revealed the relatedness among the species. The G+C content of each species was also evaluated and was found to be in range of 54.87 to 55.54%.Conclusion: The study of isolates confirmed that the isolated Bacillus sp. to be a true thermophile and could be a source of various thermostable exozymes which can be exploited for pharmaceutical and industrials applications. Much detailed study of the isolates can Int J Appl Sci Biotechnol, Vol 3(3): 392-397

What controls the diversity of life? Is it the randomness of historical contingency or the deterministic constraints of adaptive design? Several recent studies of colonial animals have suggested that nonrandom macroevolutionary patterns are common in the fossil record. These patterns generally consist of long-term, interspecific improvements in the biomechanical design and/or ecological success of some lineages over others. The problem with these previous studies is that they simply censused morphology through time. That is, they read the stratigraphic record directly. What this study does is examine macroevolutionary morphologic patterns within a phylogenetically well-defined lineage of lower Paleozoic trepostome bryozoans.17 species of ramose trepostomes ranging from the Middle Ordovician to the Upper Silurian were examined. 16 ingroup species were analyzed: 7 species of halloporids (including species belonging to Hallopora, Parvohallopora, and Tarphophragma) and 8 species of bimuroporids (including species belonging to Bimuropora and Champlainopora). Two species of Eridotrypa were chosen for the outgroup. These ingroup taxa were selected as they are united by several synapomorphs strongly suggesting this lineage is monophyletic. Phylogenetic hypotheses were constructed from a cladistic analysis of 40 multistate, qualitative, morphologic characters using outgroup analysis and the parsimony principle. This led to a single most parsimonious cladogram of 78 steps with a consistency index of 0.641.Within this lineage, species were examined for changes in the distribution of calcite across the colony by measuring a single, continuously varying, quantitative, morphologic character. Calcite distribution across cylindrical branching colonies has been shown theoretically by Wainwright et al. and empirically by Cheetham to affect branch strength. Calcite distribution was measured by exozone width. The exozone is the peripheral part of the colony where most of the calcite is precipitated due to the thick zooecial walls there. Exozone width is proportional to branch strength. This morphometric character is relatively independent of the qualitative characters used in the cladistic analysis.Exozone width was measured using a personal computer-based, digital, image analysis system. To minimize operator and system error as well as to ensure that intracolony and intraspecific variation was sufficiently represented, exozone width was measured 10 times per colony. These values were then averaged to obtain colony means. The colony means were averaged to obtain species means. A total of 261 colonies belonging to the 17 species were measured.Macroevolutionary patterns were quantified by comparing the mean exozone width for each species to 3 different evolutionary metrics derived from the most parsimonious cladogram: number of branching events each species has undergone from the base of the cladogram, number of character state transformations each species has undergone from the base of the cladogram, and the mean manhattan distance between each species and the 2 outgroup species.Regardless of which measure of evolution is chosen, the results indicate that exozone width has significantly increased through this lineage. Linear regressions revealed that these 3 trends are significant at P = 0.05. That is, the evolutionary history of this lineage reveals a progressive, nonrandom increase in branch strength. Many more species within this lineage need to be analyzed. This pattern is significant with 17 species; it may or may not be when the project is completed with the estimated 100 species that will eventually be measured. It must also be remembered that the patterns observed are dependent on the cladogram used which is dependent on the characters chosen for the phylogenetic analysis. Though these results are very preliminary, it raises the interesting question of are progressive macroevolutionary patterns more common in colonial organisms.

Recognition of ontogeny within a stenolaemate bryozoan colony requires separating the individualistic aspects of a zooid's growth from those of its neighbors. The ancestrula is the only zooid within a colony that always displays a partially independent ontogeny that ceases when it starts to experience shared changes with its neighboring daughter zooids during subsequent accretionary skeletal growth.Stenolaemate astogeny (shared changes across multiple zooids during the growth of both the ancestrular zooid and its asexual descendants) includes all coordinated changes in the size, shape, number, and calcification of autozooids, polymorphs, and extrazooidal structures, as well as changes within autozooids or polymorphs, such as the formation of basal diaphragms and brown bodies. Despite the fact that many of these directional changes occur within individual zooids, they are not part of ontogeny because they are taking place simultaneously across zooids that share a common skeleton, extrazooidal tissues, and pseudocoelomic spaces.Shared directional multizooidal changes occurring during colony growth provide a confirmatory test for the existence of astogeny. Astogeny was statistically evaluated in 6–15 characters measured within the exozones of four colonies of Tabulipora carbonaria (Worthen in Worthen and Meek, 1875). Statistically significant (at P ≤ 0.05) directional changes took place across growth stages within the exozone in the following morphometric characters: zooecial density, zooecial wall surface area, acanthostyle density, zooecial wall thicknesses, maximum acanthostyle diameters, and intrazooecial diaphragm abundances. Overall, earlier exozonal growth stages differ statistically from those of the later exozone, with characteristics of intermediate growth stages intergrading between the two. Discriminant function analysis segregated intervals of exozonal growth into early-, intermediate-, and late-stage clusters, confirming patterns delineated by univariate statistical tests.Based on these exozonal growth patterns, heterochronic changes in exozone astogeny characterized evolution within and across species of Tabulipora. Onshore populations of T. carbonaria were astogenetically progenetic relative to offshore ones along an environmental gradient across Kansas, whereas local populations became temporally more hypermorphic in a short-term stratigraphic succession of similar environments. Tabulipora carbonaria originated by astogenetic recapitulation in populations of its probable ancestor, T. ramosa. Therefore, speciation, microevolution, and clinal variation in Tabulipora all involved heterochronic modifications of exozone astogeny.

In this work, we studied some ecological and biochemical characteristics (cultural, morphological, physiological and biochemical properties, extracellular enzymatic activity, bacterial growth in various ranges of temperatures, pH, NaCl concentrations, resistance to heavy metals) of 14 strains of thermophilic bacteria taken from thermal waters and microbial mats of Kamchatka Dachnie springs. On selective environment the strains formed mostly transparent, beige, white-gray, brown, yellow and pink colonies. It was shown that the isolates were represented most of all by asporogenic, catalase-positive and oxidase-positive, mobile, gram-negative rods of various sizes. Most of the cultures were capable of forming exozymes, especially amylases and proteinases. The strains grew in a wide range of temperatures (3550C), pH (58), NaCl concentration (15%) and were characterized by increased resistance to a number of heavy metals. 4 strains of thermophilic heterotrophic bacteria exhibited multiple multidrug resistance to metals. They were identified and it was shown that the isolates were represented by bacteria of the genus Bacillus sp. (№ 19), Thermus sp. (№ 40), Pseudomonas sp. (№ 15), Rhizobium sp. (№ 125). The strains with high enzymatic activity and increased resistance to pollutants are promising for use in the field of ecology for cleaning contaminated environmental objects.

Аn attempt is made in this work to establish correlations between the level of enzymatic activity of the soil and factors of an abiotic nature. It was found that the activity of invertase and urease in the soils of more southern territories is higher than that of northern ones. In soils with a pH value of the soil environment close to neutral reaction, the level of enzymatic activity is higher than in more acidic ones. The most sensitive to soil acidity among the studied urease enzymes. In relation to zinc, an inverse correlation was established between its content in the soil and the level of invertase activity. According to the degree of resistance to salinity, the enzymes are arranged in the following order: catalase> invertase> urease. An increase in the mineralization of the soil solution leads to an increase in the activity of urease. Of the studied enzymes, the most labile are representatives of the class of hydrolases (invertase and urease), less labile are oxidoreductases (catalase). Thus, when diagnosing the state of the soil by the level of enzymatic activity, it is necessary to take into account the main abiotic factors affecting the activity of enzymes: the average annual air temperature; pH and mineralization of the soil solution; the content of substances that inhibit microorganisms and block exozymes.

All erect, branching (ramose) organisms adhere to the same fundamental geometric growth law: the rate of distal growth decreases away from the branch axis. Regardless of the phylogenetic history of an organism, the formation of cylindrical branches requires adherence to this law. In colonial ramose organisms such as trees, corals, and bryozoans, this law poses a problem. How do colonies coordinate the growth rates of the individual modules to produce an integrated branch? This question is addressed in the context of three Ordovician and three Devonian species of ramose trepostome bryozoans. Using remnant growing tips in the endozone as isochronous surfaces, relative rates of skeletal secretion among zooids were measured. Measurements of skeletal and void spaces across a colony branch enabled calculation of the volume of skeletal material secreted by zooids between successive remnant growing tips. Results indicate that rate of skeletal secretion systematically decreases from the branch axis outward into the exozone. This suggests that zooid morphogenesis is controlled to a certain degree by the colony. Colonial control over zooidal growth rates in turn regulates the shape of the colony.

Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lucia Tomášková Supervisor: prof. PharmDr. Tomáš Šimůnek, Ph.D. Title of diploma thesis: Study of exosomes as a drug delivery system in the treatment of glioblastoma Central nervous system disorders are among the most serious diseases affecting humans. They affect not only the patient's life, but also his/her surroundings. Therefore, their therapy, whether at the level of complete cure or alleviation of accompanying symptoms, is a challenge for scientific research. In our research, we focused on glioblastoma multiforme, a brain cancer not yet treatable. The main drawback in therapy is overcoming the blood-brain barrier. Exosomes, such as the body's natural nano-vesicles, have been shown to be a suitable system for delivering drugs to brain tissue. Our research has shown that by a suitable method we are able to obtain sufficient quality exosomes from macrophage and fill them very efficiently with antitumor agents paclitaxel, doxorubicin and temozolomide, while the delivered substances show higher efficacy and fewer side effects than the free form.

Extracellular vesicles (exosomes) are the subject of current nephrology proteomics research as they are considered as a promising source of potential biomarkers of kidney disease. This work is focused on discovery of the most appropriate procedure for the urinary exosomes isolation. We have compared already described methods, based on different physicochemical principles of isolation: hydrostatic filtration dialysis (HFD), differential ultracentrifugation, ultrafiltration through a 100 kDa filter, or sample precipitation with Total Exosome Isolation (from urine) kit. Characterization of individual isolated exosomal fractions was performed using SDS-PAGE method (presence of contaminating proteins), western blot analysis (detection of exosomal markers TSG101, alix), nanoparticle tracking analysis (NTA, vesicle size and concentration) or transmission electron microscopy (TEM, vesicles morphology). Due to the presence of contaminating proteins in urine samples, which could distort the results of subsequent proteomic assays, the conditions for the cleavage of undesirable proteins by proteinase K prior to their own isolation were optimized. It has been found that the best yield and purity of the isolated exosomal fractions were provided by a process combining HFD with differential ultracentrifugation...

Pregnancy-related complications such as gestational hypertension, preeclampsia, fetal growth restriction, gestational diabetes mellitus, spontaneous preterm birth, and preterm premature rupture of membranes may have severe consequences for both the mother and the child. The development of reliable early screening methods for pregnancy-related complications has therefore been a long-term goal of obstetrics. New possibilities for prenatal diagnostics have opened with the discovery of circulating microRNAs in maternal plasma. MicroRNAs are short, noncoding, 21 to 23 nucleotides long, single-strand RNAs whose main function is to regulate gene expression. During pregnancy, both common and unique miRNAs are expressed by the placenta, amongst them the miRNAs of the C19MC cluster. Several C19MC miRNAs have been shown to display a different expression profile associated with certain pregnancy-related complications. This thesis identifies the plasma exosomal profiles of six C19MC miRNAs (miR-516-5p, miR-517-5p, miR-518b, miR-520a-5p, miR-520h, and miR-525-5p) in patients in their first trimester of gestation who later developed pregnancy-related complications, and compares them with profiles in patients with normal pregnancies.