A Contemporary Appraisal on Impending Industrial and Agricultural Applications of Thermophilic-Recombinant Chitinolytic Enzymes from Microbial Sources.

The ability of chitinases to degrade the second most abundant polymer, chitin, into potentially useful chitooligomers and chitin derivatives has not only rendered them fit for chitinous waste management but has also made them important from industrial point of view. At the same time, they have also been recognized to have an imperative role as promising biocontrol agents for controlling plant diseases. As thermostability is an important property for an industrially important enzyme, various bacterial and fungal sources are being exploited to obtain such stable enzymes. These stable enzymes can also play a role in agriculture by maintaining their stability under adverse environmental conditions for longer time duration when used as biocontrol agent. Biotechnology has also played its role in the development of recombinant chitinases with enhanced activity, thermostability, fungicidal and insecticidal activity via recombinant DNA techniques. Furthermore, a relatively new approach of generating pathogen-resistant transgenic plants has opened new ways for sustainable agriculture by minimizing the yield loss of valuable crops and plants. This review focuses on the potential applications of thermostable and recombinant microbial chitinases in industry and agriculture.

Biotechnological Eminence of Chitinases: A Focus on Thermophilic Enzyme Sources, Production Strategies and Prominent Applications.

BACKGROUND: Chitin, the second most abundant polysaccharide in nature, is a constantly valuable and renewable raw material after cellulose. Due to advancement in technology, industrial interest has grown to take advantage of the chitin. OBJECTIVE: Now, biomass is being treated with diverse microbial enzymes or cells for the production of desired products under best industrial conditions. Glycosidic bonds in chitin structure are degraded by chitinase enzymes, which are characterized into number of glycoside hydrolase (GHs) families. METHODS: Thermophilic microorganisms are remarkable sources of industrially important thermostable enzymes, having ability to survive harsh industrial processing conditions. Thermostable chitinases have an edge over mesophilic chitinases as they can hydrolyse the substrate at relatively high temperatures and exhibit decreased viscosity, significantly reduced contamination risk, thermal and chemical stability and increased solubility. Various methods are employed to purify the enzyme and increase its yield by optimizing various parameters such as temperature, pH, agitation, and by investigating the effect of different chemicals and metal ions etc. Results: Thermostable chitinase enzymes show high specific activity at elevated temperature which distinguish them from mesophiles. Genetic engineering can be used for further improvement of natural chitinases, and unlimited potential for the production of thermophilic chitinases has been highlighted due to advancement in synthetic biological techniques. Thermostable chitinases are then used in different fields such as bioremediation, medicine, agriculture and pharmaceuticals. CONCLUSION: This review will provide information about chitinases, biotechnological potential of thermostable enzyme and the methods by which they are being produced and optimized for several industrial applications. Some of the applications of thermostable chitinases have also been briefly described.

MicroRNA, a Promising Biomarker for Breast and Ovarian Cancer: A Review.

BACKGROUND: MicroRNAs (miRNA) belong to the substantial class of posttranscriptional gene regulators with decisive functions in typical cellular and disease progressions. They are short RNA molecules that are not translated into proteins but bind to the complementary sites of various mRNAs, thus blocking them and leading to translational inhibition. OBJECTIVE: These miRNA molecules act as signatory molecules or biomarkers for various types of malignancies. Different miRNAs are involved in different cancer-linked pathways depending on the nature, stage, and kind of cancer. The objective of this article is to discuss and review the role and significance of various miRNAs in two of the most prominent cancers; breast and ovarian cancer. METHODS: The role of miRNAs in the instigation, propagation, and metastasis of melanoma has been elucidated. RESULTS: This article focuses on the up- and down-regulation of various miRNAs in breast and ovarian cancer, with stress on diversity in their occurrence and specificity in their threshold levels in certain types and stages of cancer, suggesting their potential role as an effective diagnostic and treatment tool for cancer. CONCLUSION: Considering all the aforementioned information, it can be concluded that miRNAs can act as potential biomarkers for the diagnosis of breast and ovarian cancers.

Next-generation sequencing to elucidate adaptive stress response and plantaricin genes among Lactobacillus plantarum strains.

Aim: The objective of this study was to identify the genes involved in plantaricin synthesis and adaptive stress response in four Lactobacillus plantarum strains (AS-6, AS-8, AS-9 and AS-10) and one Lactobacillus paraplantarum strain (AS-7) for their usage in medicine and industry. Materials & methods: Whole genomes of these strains were sequenced by a high-throughput sequencing technique known as next-generation sequencing via Ilumina MiSeq platform and the genes were identified by using various bioinformatics tools and software. Results: Plantaricin genes (plnD, plnE, plnF, plnG, plnI) and genes regulating response to temperature, pH, bile salt, osmotic and oxidative stress were identified in all strains. Conclusion: Lactobacilli could be an option to combat antimicrobial resistance and might replace harmful antibiotics in future.

Production and characterization of a novel thermo- and detergent stable keratinase from Bacillus sp. NKSP-7 with perceptible applications in leather processing and laundry industries.

Keratinase has the ability to degrade the recalcitrant keratinous wastes that cannot be degraded by conventional proteases. The present study describes a novel hyperstable keratinolytic enzyme from Bacillus sp. NKSP-7, which has excellent efficiency of keratin-feather biodegradation, washing and dehairing. The production of extracellular keratinase was improved by 3.02-fold through optimization of various parameters. Purified keratinase (25 kDa) showed optimal activity at 65 °C and pH 7.5, and displayed stability over a range of pH (5.5-9.5) and temperature (20-60 °C) for 8 h. No conspicuous effect was perceived with various chemicals and organic solvents, however, the catalytic efficiency was enhanced in the presence of Ca2+, Cd2+, Na+, Mn2+, sodium sulfite, and ß-mercaptoethanol. The enzyme was completely inhibited by phenylmethanesulfonyl fluoride (PMSF), suggesting that this keratinase belongs to serine protease family. It displayed prodigious stability and compatibility to salinity and commercial detergents. Enzyme exhibited great substrate specificity but high affinity was observed with keratin-rich substrates. Crude and purified keratinase revealed perceptible potential for destaining of blood-stained fabric (10 min), and dehairing of hide (8 h) without any damage. All these auspicious features make this enzyme a promising candidate for various industrial applications, especially in keratin-waste management, detergent formulations and leather processing.

Keratinolytic enzyme-mediated biodegradation of recalcitrant poultry feathers waste by newly isolated Bacillus sp. NKSP-7 under submerged fermentation.

Microbial and enzymatic degradation of keratin waste is more preferred over various conventional approaches which are costly and not environmentally suitable. Diverse niches are auspicious for the discovery of new microorganisms. To discover novel keratinolytic bacteria, 60 isolates from different poultry dumping sites were initially screened, and among these found a potent keratinolytic isolate (NKSP-7) that displayed higher feather-degrading ability. The selected isolate was identified as Bacillus sp. NKSP-7 based on 16S rDNA sequencing as well as physiochemical and morphological characteristics. The strain NKSP-7 showed complete hydrolysis of native chicken feathers (10 g/L) in nutrient medium after 24 h of incubation at 37 °C under agitation (150 rev/min) and produced thermostable extracellular keratinase. The crude enzyme displayed maximal keratinolytic activity (34.7 U/mL) in phosphate buffer of pH 7.0, and at 60 °C using keratin azure as a substrate. Keratinolytic enzyme showed stability at 20-65 °C for 4 h over the pH range of 5.5-8.0. No obvious inhibitory influence was perceived by cations, organic solvents, EDTA, and detergents. Whereas, enzyme activity was enhanced by adding ß-mercaptoethanol, Na+, Cd2+, and Mn2+. All these notable features of keratinase make it a promising candidate for various industrial applications especially for dehairing process in leather industry, bioconversion of poultry waste, and in detergents formulations.