Bioconversion of agro-industrial wastes for the production of fibrinolytic enzyme from Bacillus halodurans IND18: Purification and biochemical characterization

Background: Agro-wastes were used for the production of fibrinolytic enzyme in solid-state fermentation. The process parameters were optimized to enhance the production of fibrinolytic enzyme from Bacillus halodurans IND18 by statistical approach. The fibrinolytic enzyme was purified, and the properties were studied. Results: A two-level full factorial design was used to screen the significant factors. The factors such as moisture, pH, and peptone were significantly affected enzyme production and these three factors were selected for further optimization using central composite design. The optimum medium for fibrinolytic enzyme production was wheat bran medium containing 1% peptone and 80% moisture with pH 8.32. Under these optimized conditions, the production of fibrinolytic enzyme was found to be 6851 U/g. The fibrinolytic enzyme was purified by 3.6-fold with 1275 U/mg specific activity. The molecular mass of fibrinolytic enzyme was determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis, and it was observed as 29 kDa. The fibrinolytic enzyme depicted an optimal pH of 9.0 and was stable at a range of pH from 8.0 to 10.0. The optimal temperature was 60°C and was stable up to 50°C. This enzyme activated plasminogen and also degraded the fibrin net of blood clot, which suggested its potential as an effective thrombolytic agent. Conclusions: Wheat bran was found to be an effective substrate for the production of fibrinolytic enzyme. The purified fibrinolytic enzyme degraded fibrin clot. The fibrinolytic enzyme could be useful to make as an effective thrombolytic agent.

Biosynthesis and characterization of keratinolytic protease from actinobacterium sp. in solid state culture.

The aim of this study was to use agro-industrial residues for the production of a halotolerant keratinolytic- protease by Actinobacterium sp. in solid-state fermentation. Among various agro-industrial residues that were evaluated, apple pomace supported maximum protease production (8400 U/g material). The optimum conditions required for enzyme production were a fermentation period of 72 h, 10% (w/v) NaCl, pH 7.0, 120% (v/w) moisture and 10% (v/w) inoculum. The enzyme exhibited activity to a range of pH (7.0-9.0) and temperature (30-45°C), with optima at 8.0 and 40 °C, respectively. Most of the divalent ions tested stimulated the protease activity and Ca2+ ion was required for its activity and stability. The enzyme was widely active at the range of NaCl concentration (5%-15%, w/v) and effectively degraded chicken feather. This protease could be useful in fish sauce fermentation and also in feed industry.

Tannase production by Penicillium atramentosum KM under SSF and its applications in wine clarification and tea cream solubilization

Tannin acyl hydrolase commonly known as tannase is an industrially important enzyme having a wide range of applications, so there is always a scope for novel tannase with better characteristics. A newly isolated tannase-yielding fungal strain identified as Penicillium atramentosum KM was used for tannase production under solid-state fermentation (SSF) using different agro residues like amla (Phyllanthus emblica), ber (Zyzyphus mauritiana), jamun (Syzygium cumini), Jamoa (Eugenia cuspidate) and keekar (Acacia nilotica) leaves. Among these substrates, maximal extracellular tannase production i.e. 170.75 U/gds and 165.56 U/gds was obtained with jamun and keekar leaves respectively at 28ºC after 96 h. A substrate to distilled water ratio of 1:2 (w/v) was found to be the best for tannase production. Supplementation of sodium nitrate (NaNO3) as nitrogen source had enhanced tannase production both in jamun and keekar leaves. Applications of the enzyme were studied in wine clarification and tea cream solubilization. It resulted in 38.05 percent reduction of tannic acid content in case of jamun wine, 43.59 percent reduction in case of grape wine and 74 percent reduction in the tea extract after 3 h at 35ºC.

Studies on sorption properties of pathogens on natural materials.

Presence of pathogens in high numbers in waste water is a cause of concern. Techno economic feasibility has restricted the conventional and non conventional treatment approaches for pathogen removal. Despite prolific use, carbon adsorption technology remains an expensive treatment process. The present study investigates the use of rice husk (RH), saw dust (SD), groundnut shells (GS) as natural agro-residues and partially weathered deccan trap basalt (PWDTB) for their sorption capacities and desorption pattern for two indicator organisms viz. Escherichia coli K12 and Staphylococcus aureus. Sorption experiments were carried out at flow-rate of 1.5 bed volumes per hour (bv hr-1) for cell suspension volume of 4, 8, 16 and 32 bed volumes. PWDTB have shown high sorption coefficient and log removal for E. coli K12 whereas GS have shown high sorption coefficient and log removal for S. aureus. PWDTB have shown maximum desorption constant and log retention for E. coli K12 whereas GS have shown maximum desorption constant and log retention for S. aureus during desorption experiment. Retention pattern suggest that adsorption is partially irreversible for almost all the materials used. It suggest that PWDTB in combination with RH and / GS could help in removal of pathogens from waste water.

Avaliação da produção de Pleurotus sajor-caju (Fr.) singer utilizando resíduo do beneficiamento têxtil do algodão como substrato / Evaluation of the production of Pleurotus sajor-caju in cotton textille mill waste

Realizou-se um ensaio objetivando investigar a viabilidade da produção do cogumelo comestível Pleurotus sajor-caju (Fr.) Singer em substratos cuja principal fonte de carbono fosse o resíduo do beneficiamento têxtil do algodão. Foram avaliadas duas composições de substrato: C1 (resíduo do beneficiamento têxtil do algodão, farelo de trigo, gesso e calcário) e C2 (resíduo do beneficiamento têxtil do algodão, farelo de trigo, palha de feijão, gesso e calcário). O experimento foi estabelecido em delineamento inteiramente casualizado, com nove repetições por tratamento e os dados de produção e eficiência biológica foram analisados utilizando-se o procedimento ANOVAG do pacote estatístico SAEG. O resíduo do beneficiamento têxtil do algodão, como principal ingrediente do substrato, mostrou-se eficiente para a produção do cogumelo P. sajor-caju, apresentando valores satisfatórios de produtividade (0,56 e 0,55 kg/kg substrato) e eficiência biológica (55,76 e 55,39 por cento), respectivamente para C1 e C2, podendo ser recomendado como alternativa menos onerosa de substrato comercial para a espécie Pleurotus sajor-caju.

The experiment was conducted to investigate the viability of the production of the edible fungus Pleurotus sajor-caju (Fr.) Singer in substrates in which main source of carbon was the cotton textille mill waste. Two substrates compositions were tested: C1 (cotton textille mill waste, wheat bran, plaster and limestone) and C2 (cotton textille mill waste, wheat bran, bean straw, plaster and limestone). A DIC experimental design was used, with nine repetitions for treatment and the production data and biological efficiency were analyzed being used the procedure ANOVAG of the statistical package SAEG. The cotton textille mill waste improvement, as the main ingredient of the substrate, was shown efficient for the production of the mushroom P. sajor-caju, presenting satisfactory values of productivity (0,56 and 0,55 kg/kg substratum) and biological efficiency (55,76 and 55,39 percent), respectively for C1 and C2. These results showed us that cotton textille mill waste, could be recommended as less onerous alternative of commercial substrate for the species Pleurotus sajor-caju.