Extraction of chitosan from biologically-derived chitin by bacterial chitin deacetylase: Process optimization and product quality assessment.

Commercial chitosan manufacturing process relies on strong chemical treatment on chitin that generates chitosan with undesirable properties and leads to environmental pollution. To overcome the adverse consequences, enzymatic chitosan preparation from chitin was undertaken in the current study. A potent chitin deacetylase (CDA) producing bacterial strain was screened and subsequently identified as Alcaligens faecalis CS4. After optimization 40.69 U/mL of CDA production was achieved. By treating the organically extracted chitin with partially purified CDA chitosan yield of 19.04 % was attained having 71 % solubility, 74.9 % degree of deacetylation, 21.16 % crystallinity index, 246.4 kDa molecular weight and 298 °C highest-decomposition temperature. FTIR and XRD analysis revealed characteristics peaks respectively within 870-3425 cm-1 wavenumber and 10°-20°, for enzymatically and chemically extracted (commercial) chitosan that endorses their structural similarity which validated through electron microscopic study. At 10 mg/mL chitosan concentration 65.49 % DPPH radical scavenging activity endorsed its antioxidant potential. Minimum inhibitory concentration of chitosan was 0.675, 1.75, 0.33 and 0.75 mg/mL for Streptococcus mutans, Enterococcus faecalis, Escherichia coli and Vibrio sp., respectively. Mucoadhesiveness and cholesterol binding properties were also exhibited by extracted chitosan. The present study opens a new vista for eco-friendly extraction of chitosan from chitin that is proficient and sustainable in environmental perspective.

Production of glutathione from probiotic Bacillus amyloliquefaciens KMH10 using banana peel extract.

Glutathione, a tri-peptide (glutamate-cysteine-glycine) with the thiol group (-SH), is most efficient antioxidative agent in eukaryotic cells. The present study aimed to isolate an efficient probiotic bacterium having the potential to produce glutathione. The isolated strain Bacillus amyloliquefaciens KMH10 showed antioxidative activity (77.7 ± 2.56) and several other essential probiotic attributes. Banana peel, a waste of banana fruit, is chiefly composed of hemicellulose with various minerals and amino acids. A consortium of lignocellulolytic enzyme was used for the saccharifying banana peel to produce 65.71 g/L sugar to support the optimal glutathione production of 181 ± 4.56 mg/L; i.e., 1.6 folds higher than the control. So, the studied probiotic bacteria could be an effective resource for glutathione; therefore, the stain could be used as natural therapeutics for the prevention/treatment of different inflammation-related gastric ailments and as an effective producer of glutathione using valorized banana waste that has excellent industrial relevance.

Efficient valorization of feather waste by Bacillus cereus IIPK35 for concomitant production of antioxidant keratin hydrolysate and milk-clotting metallo-serine keratinase.

Keratinase production by Bacillus cereus IIPK35 was investigated under solid-state fermentation (SSF) and the maximum titer of 648.28 U/gds was revealed. Feather hydrolysates obtained from SSF exhibited paramount antioxidant properties in ABTS [2,2'-azinobis-(3-ethylbenzothiazoline)-6-sulfonic acid], FRAP [Ferric ion reducing antioxidant power], and DPPH [2,2,-Diphenyl-1-picrylhydrazyl] assay. The keratinase was purified up to homogeneity have a molecular weight of 42 kDa, and showed its stability between pH 6.5-10.0 and temperature 35-60 °C with optimum enzyme activity at pH 9.0 and 55 °C. The catalytic indices viz. Km of 9.8 mg/ml and Vmax of 307.7 µmol/min for keratin were determined. Besides keratin, the enzyme displayed broad and proteolytic activity towards other proteinaceous substrates such as casein, skim milk, gelatin, and bovine serum albumin. Pure keratinase activity was stimulated in presence of Ca2+ and Mg2+ ions, while it was strongly inhibited by both iodoacetamide and EDTA, indicating it to be a metallo-serine protease in nature. Circular dichroism study endorses the structural stability of the secondary structure at the said range of pH and temperature. The IIPK35 keratinase is non-cytotoxic in nature, shows remarkable storage stability and is stable in presence of Tween 80, Triton X 100, and sodium sulfite. Furthermore, it showed excellent milk clotting potential (107.6 Soxhlet Unit), suggesting its usefulness as an alternative milk clotting agent in the dairy industry. This study unlocks a new gateway for keratinase investigation in SSF using chicken feathers as substrate and biochemical and biophysical characterization of keratinase for better understanding and implication in industrial applications.

Application of Phytochemicals To Combat Fungal Pathogens of Pulses: An Approach toward Inhibition of Fungal Propagation and Invasin Activity.

The present study represented an innovative strategy for inactivating the secreted invasins (lignocellulolytic enzymes) of fungal phytopathogens using natural phytochemicals to combat fungal infection to the pulses. A fungal pathogen (Aspergillus niger SKP1) was isolated from the white lentil (Vigna mungo), which has the ability to synthesize different lignocellulolytic enzymes. An in silico docking study elucidated that quercetin, naringin, epigallocatechin gallate, curcumin, and cinnamic acid were the prime efficient phytochemicals to inhibit the activity of fungal invasive enzymes like endoglucanase, endo-1,4-ß-xylanase, and glucoamylase. Considering this observation, extracted phytochemicals in different mixtures were applied to prevent growth of the isolated pathogen under in situ experimental studies. The minimal inhibitory concentrations (MIC50) and minimal fungicidal concentration (MFC50) values of the first mixture (naringenin, epicatechin gallate, and cinnamic acid) and second mixture (quercetin and curcumin) were 170 and 220 mg/L and 320 and 380 mg/L, respectively. The studied phytochemicals were established to be cytosafe when compared to the commercial fungicides. The seeds of the white lentil were subjected to 1 year of long-term storage with the two aforementioned combinatorial phytochemicals. Subsequent morphological and physiological analyses revealed the complete protection of the stored seeds from the fungal infection. The present work has enough potentiality for the storage of pulses using natural preservatives that circumvent the adverse effect of the chemical preservatives on the ecosystem.

Antiobesity, Antihyperglycemic, and Antidepressive Potentiality of Rice Fermented Food Through Modulation of Intestinal Microbiota.

The present study has been aimed at evaluating the antiobesity, antihyperglycemic, and antidepressive potentials of Asparagus racemosus starter-based rice fermented foods. High-throughput NGS technology has revealed a number of bacterial genera in the prepared fermented rice, such as Lactobacillus (29.44%), Brevundimonas (16.21%), Stenotrophomonas (6.18%), Pseudomonas (3.11%), Bacillus (2.88%), and others (<2%). Eight-week administration of rice fermented food has increased food intake, whole-body weight, organ weight, different fat masses, serum lipid profiles, and histology of liver and adipose tissues in HFD-induced obese mice. In addition, upregulation of fatty acid oxidation and downregulation of adipocytogenesis- and lypogenesis-related genes along with the expression of their regulatory nuclear factors such as PPARα, PPARγ, PPARδ, and SREBP-1c have also been noted. Moreover, fermented food decreases fasting blood glucose level and improves glucose and insulin tolerance as well as the expression of GLUT4 receptor. Antiobesity and antihyperglycemic effects are also supported by the changes in insulin, leptin, and adiponectin hormone levels. The real-time polymerase chain reaction (RT-PCR) and denaturing gradient gel electrophoresis (DGGE) analyses have clearly demonstrated the intense colonization of Bacteroides, Lactobacillus, and Bifidobacterium, as well as the suppressed growth rate of γ- and δ-Proteobacteria and Firmicutes in the gut after fermented food intake. In the intestine, the latter group of microorganisms possibly modulate short-chain fatty acid (SCFA) levels such as acetate, butyrate, and propionate more than twofold. The impairment of memory-learning and anxiety-like obesity-associated cognitive phenotypes is mitigated significantly (p < 0.01) by fermented food as well. Thus, the formulated fermented food could be used as a natural therapeutic to alleviate obesity and its associated psychological and pathophysiological ailments.

Preparation of rice fermented food using root of Asparagus racemosus as herbal starter and assessment of its nutrient profile.

The popularity of traditional fermented food products is based on their healthiness. The addition of a starter brings consistent, desirable, and predictable food changes with improved nutritive, functional, and sensory qualities. The addition of a mixture of plant residues as a starter or source of microbes is an age-old practice to prepare traditional fermented food and beverages, and most of the reported data on traditional foods were based on the analysis of the final product. The contribution of an individual starter component (plant residue) is not experimentally substantiated for any traditional fermented food, but this data are very essential for the formulation of an effective starter. In this study, Asparagus racemosus, which used as a common ingredient of starter for preparation of rice fermented food in the Indian sub-continent, was used as a starter for the preparation of rice fermented food under laboratory scale, and its microbial and nutrient profile was evaluated. The fermented product was a good source of lactic acid bacteria, Bifidobacterium sp., yeast, etc. The food product was acidic and enriched with lactic acid and acetic acid with titratable acidity of 0.65%. The content of protein, fat, minerals, and vitamins (water-soluble) was considerably improved. Most notably, oligosaccharide (G3-matotriose), unsaturated fatty acids (ω3, ω6, ω7, and ω9), and a pool of essential and non-essential amino acids were enriched in the newly formulated food. Thus, the herbal starter-based rice fermented food would provide important macro- and micronutrients. They could also deliver large numbers of active microorganisms for the sustainability of health. Therefore, the selected plant part conferred its suitability as an effective starter for the preparation of healthier rice-based food products.

Tailoring in fungi for next generation cellulase production with special reference to CRISPR/CAS system.

Cellulose is the utmost plenteous source of biopolymer in our earth, and fungi are the most efficient and ubiquitous organism in degrading the cellulosic biomass by synthesizing cellulases. Tailoring through genetic manipulation has played a substantial role in constructing novel fungal strains towards improved cellulase production of desired traits. However, the traditional methods of genetic manipulation of fungi are time-consuming and tedious. With the availability of the full-genome sequences of several industrially relevant filamentous fungi, CRISPR-CAS (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein) technology has come into the focus for the proficient development of manipulated strains of filamentous fungi. This review summarizes the mode of action of cellulases, transcription level regulation for cellulase expression, various traditional strategies of genetic manipulation with CRISPR-CAS technology to develop modified fungal strains for a preferred level of cellulase production, and the futuristic trend in this arena of research.

Extraction of chitin from Litopenaeus vannamei shell and its subsequent characterization: an approach of waste valorization through microbial bioprocessing.

Chemical extraction of chitin is very hazardous and costly which can be overwhelmed by microbial bioprocessing. In this study, potent protease and lactic acid-producing bacteria were screened and identified as Alcaligens faecalis S3 and Bacillus coagulans L2, respectively. Productions of protease and lactic acid by the respective bacterial strains were optimized. The shell of Litopenaeus vannamei was sequentially treated with the partially purified protease and lactic acid and the treatment conditions were optimized for betterment of chitin yield. Spectral characterization by SEM-EDS, IR, XRD, NMR, XPS and thermal characterization by TG and DTG analysis of the extracted chitin was made and compared with commercial one. It was revealed that both the chitin have similar characteristics. Therefore, it can be articulated that chitin can be extracted from crustacean shells in pure form by microbial bioprocessing which will be a good catch for biorefinary industries for chitin extraction through greener route.

Microbial decomposition of crustacean shell for production of bioactive metabolites and study of its fertilizing potential.

Crustacean shell waste disposal is considered as biggest problem in seafood processing centers. Incineration and landfilling are the commonest ways of disposal of the waste which causes environmental pollution. Microbial bio-conversion is one of the promising approaches to minimize the wastes by utilizing the same for deriving different value added metabolites. In this perspective, chitinase- and protease-producing bacterial strains were isolated from shrimp culture pond, and the potent isolate was subsequently identified as Alcaligenes faecalis SK10. Fermentative optimization of the production of chitinase (85.42 U/ml), protease (58.57 U/ml), and their catalytic products, viz., N-acetylamino sugar (84 µg/ml) and free amino acids (112 µg/ml), were carried out by utilizing shrimp and crab shell powder as principal substrate. The fermented hydrolysate (FH) was subsequently applied to evaluate its potential to be a candidate fertilizer for the growth of leguminous plant Pisum sativum and Cicer arietinum, and the results were compared with chitin, chitosan, and commercial biofertilizer amended group. The results revealed that FH have paramount potential to improve plants morpho-physiological parameters like stem and root length, chlorophyll, cellular RNA, protein content, and soil physico-chemical parameters like total nitrogen, magnesium, calcium, phosphorus, and potassium significantly (p < 0.05). Moreover, the application of FH also selectively encouraged the growth of free-living nitrogen-fixing bacteria, Rhizobium, phosphate-solubilizing bacteria in the soil by 4.82- and 5.27-, 5.57- and 4.71, and 7.64- and 6.92-fold, respectively, in the rhizosphere of P. sativum and C. arietinum, which collectively is a good sign for an ideal biofertilizer. Co-supplementation of FH with commercial PGPR-biofertilizer significantly influenced the morpho-physiological attributes of plant and physico-chemical and microbial attributes of soil. The study validated proficient and sustainable utilization of fermented hydrolysate of waste crustacean shell as biofertilizer.

Multi-Response Optimization of Granaticinic Acid Production by Endophytic Streptomyces thermoviolaceus NT1, Using Response Surface Methodology.

Streptomyces thermoviolaceus NT1, an endophytic isolate, was studied for optimization of granaticinic acid production. It is an antimicrobial metabolite active against even drug resistant bacteria. Different media, optimum glucose concentration, initial media pH, incubation temperature, incubation period, and inoculum size were among the selected parameters optimized in the one-variable-at-a-time (OVAT) approach, where glucose concentration, pH, and temperature were found to play a critical role in antibiotic production by this strain. Finally, the Box-Behnken experimental design (BBD) was employed with three key factors (selected after OVAT studies) for response surface methodological (RSM) analysis of this optimization study.RSM analysis revealed a multifactorial combination; glucose 0.38%, pH 7.02, and temperature 36.53 °C as the optimum conditions for maximum antimicrobial yield. Experimental verification of model analysis led to 3.30-fold (61.35 mg/L as compared to 18.64 mg/L produced in un-optimized condition) enhanced granaticinic acid production in ISP2 medium with 5% inoculum and a suitable incubation period of 10 days. So, the conjugated optimization study for maximum antibiotic production from Streptomyces thermoviolaceus NT1 was found to result in significantly higher yield, which might be exploited in industrial applications.

Appraisal of antioxidant, anti-hemolytic and DNA shielding potentialities of chitosaccharides produced innovatively from shrimp shell by sequential treatment with immobilized enzymes.

Chitosaccharides (CS) of varied size were prepared from shrimp shell through sequential catalysis, using crude protease and chitinase enzymes immobilized on agar beads. In the optimized state, immobilization yield and activity yield for protease were 84% and 62%, and for chitinase were 75% and 57%, respectively. Immobilized protease and chitinase treatment improved CS yields (101 µg/ml) and retained 63% and 52% of activities after 10 reuses, respectively. Stronger radical-scavenging activity (RSA) of CS against ABTS, DPPH and hydroxyl radical was noted with EC50 values 19.1, 26.4 and 29.6 µg/ml, respectively. Peroxyl and superoxide RSAs of 96.8% and 88.6% were noticed at 70 µg/ml of CS. Singlet oxygen quenching, reducing power and ferrous ion-chelating activities of CS were also pronounced. CS reasonably reduced oxidative damage of DNA, protein and RBC by inhibiting H2O2 and AAPH radicals. Reversible CS-DNA condensation leads to DNA stabilization without changing its conformation and advocates its employment in gene therapy.

Biosynthesis, structural architecture and biotechnological potential of bacterial tannase: a molecular advancement.

Tannin-rich materials are abundantly generated as wastes from several agroindustrial activities. Therefore, tannase is an interesting hydrolase, for bioconversion of tannin-rich materials into value added products by catalyzing the hydrolysis of ester and depside bonds and unlocked a new prospect in different industrial sectors like food, beverages, pharmaceuticals, etc. Microorganisms, particularly bacteria are one of the major sources of tannase. In the last decade, cloning and heterologous expression of novel tannase genes and structural study has gained momentum. In this article, we have emphasized critically on bacterial tannase that have gained worldwide research interest for their diverse properties. The present paper delineate the developments that have taken place in understanding the role of tannase action, microbial sources, various cultivation aspects, downstream processing, salient biochemical properties, structure and active sites, immobilization, efforts in cloning and overexpression and with special emphasis on recent molecular and biotechnological achievements.

Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation.

Production and optimization of ß-N-acetyl glucosaminidase and chitinase by Ca-alginate immobilized Aeromonas hydrophila SBK1 was carried out using prawn shell as cost-effective substrate. Beads prepared with 5.0% Na-alginate (containing 2.0% colloidal chitin) and 1.0 M CaCl2 showed considerable beads integrity and supported maximum production of chitinolytic enzymes. Bead diameter, 3 mm; temperature, 35°C; pH 7.0; agitation, 90 rpm were found ideal for the maximum production of the enzymes. The fermentation and thermodynamic indices revealed the feasibility of immobilized cells over free cells for enzymes production. Reasonable amount of chitosaccharides (degree of polymerization; 1-6) accumulated in the production media which have paramount antioxidant activity. Scale up experiment was successfully carried out in 5 L fermentor. In immobilized state, the chitosaccharides yield and antioxidant activity increased about 44.76% and 22.22%, whereas specific productivity of ß-N-acetyl glucosaminidase and chitinase increased by 22.86% and 33.37% over free state. The cell entrapped beads can be reused upto ten cycles without marked loss of its biocatalytic efficiency. High level of protoplast of Aspergillus niger was generated by treating mycelia with 10 U/ml of crude chitinase after 4 h at pH 7.0 and in the temperature 35-40°C, and 67% of the protoplasts were found to be regenerated.

Thermodynamics and kinetic properties of halostable endoglucanase from Aspergillus fumigatus ABK9.

An endoglucanase from Aspergillus fumigatus ABK9 was purified from the culture extract of solid-state fermentation and its some characteristics were evaluated. The molecular weight of the purified enzyme (56.3 kDa) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, zymogram analysis and confirmed by MALDI-TOF mass spectrometry. The enzyme was active optimally at 50 °C, pH 5.0 and stable over a broad range of pH (4.0-7.0) and NaCl concentration of 0-3.0 M. The pKa1 and pKa2 of the ionizable groups of the active sites were 2.94 and 6.53, respectively. The apparent Km , Vmax , and Kcat values for carboxymethyl cellulose were 6.7 mg ml(-1), 775.4 µmol min(-1) , and 42.84 × 10(4) s(-1), respectively. Thermostability of the enzyme was evidenced by the high activation energy (91.45 kJ mol(-1)), large enthalpy for activation of denaturation (88.77 kJ mol(-1)), longer half-life (T1/2) (433 min at 50 °C), higher melting temperature (Tm ) (73.5 °C), and Q10 (1.3) values. All the characteristics favors its suitability as halotolerant and thermostable enzyme during bioprocessing of lignocellulosic materials.

Study on regulation of growth and biosynthesis of cellulolytic enzymes from newly isolated Aspergillus fumigatus ABK9.

This study was aimed to evaluate the pattern of cellulase biosynthesis from Aspergillusfumigatus ABK9 under submerged fermentation. Production was increased concomitantly with fungal growth up to 72 h and reached maximum (Xmax -6.72 g/l) with specific growth rate (mu max) of 0.126/h. Highest specific rate of enzyme production (q ) was found at initial medium pH of 5.0 and incubation temperature of 30 degrees C. At the same time, in the presence of 2-deoxy-D-glucose concentration of 0.5 mg/ml, the production of cellulolytic enzymes, viz, carboxymethyl cellulase activity (CMCase), filter paper degrading activity (FPase) and P-glucosidase activity reached maximum of 132.2, 21.3 and 28.9 U/ml, respectively. Cellulase biosynthesis was induced in respect to higher volumetric production rate (Qp), specific rate of enzymes production (qp, U/g biomass/h) and enzyme/biomass yield (YE/X) when grown in carboxymethyl cellulose in comparison to other saccharides as sole carbon source. Induction ratios (IR) of cellulases were between 12.3 and 24.4 in the presence of 1.5% (w/v) CMC in the culture media. The strain was quite resistant to catabolic repression by glucose up to 0.4% (w/v). Cellulases production was greatly influenced in the presence of yeast extract and potassium dihydrogen phosphate (KH2POA) as nitrogen and phosphate sources in the culture media. C/N ratio of 10.0 and C/P ratio of 4.0 proved to be the best for the production of enzyme cocktail. Along with the high production yield, the crude enzymes showed a promising cellulose hydrolyzing efficiency of rice straw, indicating the enzyme could be beneficial for its large scale industrial exploitation.

Exploitation of fermented shrimp-shells hydrolysate as functional food: assessment of antioxidant, hypocholesterolemic and prebiotic activities.

In the present study the bioactivities of chitooligosaccharides of fermented shrimp-shell hydrolysate (SSH) in respect to hypocholesterolemic, antioxidant and prebiotic activity were tested in male albino rat. Rats were treated with four different diets, viz., (i) cholesterol-rich (5%) basal diet (ChB), (ii) ChB+10% chitin, (iii) ChB+10% SSH and (iv) control group (without cholesterol). After 4 weeks of treatment, body mass index, liver weight, serum total cholesterol and LDL-cholesterol in groups (ii) and (iii) were decreased significantly than group (i). SSH supplementation significantly resists oxidative stress by reducing the thiobarbituric acid reactive substances and by increasing catalase, superoxide dismutase and free radical scavenging activity. The colonization of Lactobacillus and Bifidobacterium population in small and large intestine were more in group (iii) than other groups. Reduction of Clostridium perfringens population and non-significant changes of E. coli was also noted in SSH supplement group. Histological study revealed that the villus height and villus:crypt of the small intestine were increased significantly in SSH supplemented group (iii) without any diarrheal symptoms. The results demonstrated that the shrimp-shells hydrolysate has hypocholesterolemic effect, can resist lipid peroxidation and can influence the growth of health beneficial microbes, hence can be used as functional food for hypercholesterolemic patients.

Analysis of alteration of gut microbial population under the exposure of graded hyperbaric pressures: application of metagenomic approach.

Gastroenterological disorders are very common at hyperbaric conditions. The present study was conducted to find out the impact of gut flora on the gastrointestinal disorders created at such environmental circumstances. For this, male albino rat were exposed to graded hyperbaric pressures (915 and 1277 mmHg) and large intestinal content was examined for microbial composition using culture based and PCR-DGGE tools. After 30 day exposure, total aerobes (38.54 and 375.57 folds, 1.35 and 1.58 gdi) and E. coli (126.05 and 873.23 folds, 1.31 and 1.44 gdi) were increased whereas total anaerobes (7.01 x 10(4) and 8.84 x 10(3) folds, -1.56 and -1.39 gdi), Enterobacter spp. (-2.45 and -1.00 gdi) and Clostridium perfringens (12.88 and 54.16 folds, -1.38 and -1.75 gdi) were decreased significantly in respect to control after exposure of simulated hyperbaric pressures like at 915 and 1277 mmHg, respectively. Metagenomics study revealed an overall reduction in total microbial profile was noted than control at higher level hyperbaric pressure, i.e., 1277 mmHg air pressure for highest duration of exposure. Though, some new bands also appeared which indicated the expansion of dormant or new microbiota, Variation in the numbers of these newly dominated bacteria was correlated to dose and duration of hyperbaric treatment. The histological results clearly indicated that hyperbaric environment induced severe inflammation in the mucosal and submucosal layer of large intestine. Thus, the result suggest that hyperbaric pressure is an important exogenous factor that strongly modulated the intestinal morphology and microbial ecology, and induced several gastrointestinal ailments during hyperbarism.

Rapid screening of tannase producing microbes by using natural tannin

Use of natural tannin in the screening of tannase producing microbes is really promising. The present work describes about the possibility and integrity of the newly formulated method over the previously reported methods. Tannin isolated from Terminalia belerica Roxb. (Bahera) was used to differentiate between tanninolytic and nontanninolytic microbes. The method is simple, sensitive and superior for the rapid screening and isolation of tannase-producing microbes.

Tannase production by Penicillium purpurogenum PAF6 in solid state fermentation of tannin-rich plant residues following OVAT and RSM.

Tannase production by newly isolated Penicillium purpurogenum PAF6 was investigated by 'one variable at a time' (OVAT) approach followed by response surface methodology (RSM). Tannin-rich plant residues were used as supporting solid substrate and sole carbon source and, among them, tamarind seed was found to be the most favorable substrate than haritaki, pomegranate, tea leaf waste and arjun fruit. Physicochemical parameters were initially optimized using OVAT methodology and some important factors like incubation time, incubation temperature, substrate:moisture ratio as well as carbon, nitrogen and phosphate concentrations were verified with Box-Behken design of response surface methodology. Phosphate source, nitrogen source and temperature were found as the most favorable variables in the maximization of production. Tannase production was enhanced from 1.536 U/g to 5.784 U/g using tamarind seed OVAT optimization and further enhancement up to 6.15 U/g following RSM. An overall 3.76- and 4.0-fold increases in tannase production were achieved in OVAT and RSM, respectively.

Rapid screening of tannase producing microbes by using natural tannin.

Use of natural tannin in the screening of tannase producing microbes is really promising. The present work describes about the possibility and integrity of the newly formulated method over the previously reported methods. Tannin isolated from Terminalia belerica Roxb. (Bahera) was used to differentiate between tanninolytic and nontanninolytic microbes. The method is simple, sensitive and superior for the rapid screening and isolation of tannase-producing microbes.