Identification, isolation, cloning and sequencing of tannase. Although tannase production by aspergillus niger can occur in the absence of tannic acid, this fungus tolerates tannic acid concentrations as high as 20%wv, without having a deleterious effect on both the growth and enzyme production. Nowadays, tannase is produce by submerged culture smc being the enzyme obtained in an intracellular form resulting, as a consequence, in high production costs. Research on conditions of solidstate fermentation and.
Pdf optimization of tannase production by aspergillus. One variable at a time procedure was used to evaluate the effect of qualitative variables on the production of tannase from aspergillus niger van tieghem. Elbanna department of food science and technology, faculty of agriculture, alexandria university, alexandria. Production, characterization and purification of tannase from aspergillus niger article pdf available in european journal of experimental biology 25. Statistically based experimental design was employed for the optimization of fermentation conditions for maximum production of enzyme tannase from aspergillus niger. Aspergillus niger growth on polyphenolic carbon sources and optimization of the tannase production souad akroum department of applied microbiology and food sciences, faculty of natural sciences, university of jijel, ouled aissa city, jijel 18000, algeria. Statistical optimization for tannase production from. Culture conditions for the production of a tannase of aspergillus tamarii imi388810 b enemuor, s. This study was carried out to optimize the physical parameters for the maximal tannase production by aspergillus niger via solid substrate fermentation using a shake flask system. College, nanded 431601, india corresponding author abstract introduction gallic acid, a phenolic compound, chemically known as 3,4,5. Characterization of a thermostable extracellular tannase. Agitation speed fungal hyphae aspergillus niger tannase introduction the economics of a process.
Accepted april, 2009 aspergillus tamarii imi388810 b a tannic acid degrading fungus was isolated from soil inundated by effluent of a tannery at oji river local government area of enugu state, nigeria. Based on the sem and tem micrograph observation, there was. Influence of agitation speed on tannase production and. A decrease in catechin and theaflavin gallates and a significant increase in ga content and the relative. Evaluation of culture conditions for tannase production by. A study on optimization of submerged fermentative production of tannase by aspergillus niger was carried out. A tannase from aspergillus niger was used for esterification of catechin using gallic acid in ionic liquids, a process known as galloylation raab et al. Zumrut ogel august 2006, 90 pages in this study, a filamentous fungus, aspergillus niger was evaluated in terms of extracellular tannase production. Thermostable tannase from aspergillus niger application in. Pdf selection of tannase producing aspergillus niger strains. Sep 27, 2011 agitation speed was found to influence the tannase production and fungal growth of aspergillus niger fetl ft3. Production, characterization and purification of tannase. Using designed primers, pcr carried out and a 950bp expected band obtained that was ligated into cloning vector and then cloned into escherichia coli top10f in.
Bioconversion of naturally occurring tannins into a value added pharmaceutical intermediate gallic acid a new approach h. Identification, isolation, cloning and sequencing of. Extra and intracellular tannase production by aspergillus niger gh1 has been evaluated using submerged smf and solidstate fermentation ssf at different temperatures 30, 40 and 50 c. Accordingly, four isolates, which exhibited high tannase activity, were chosen to. The experiments involved in the process and conditions of solidstate fermentation and factors influencing characters of tannase. Original article statistical optimization for tannase production from aspergillus niger under submerged fermentationr.
This fungus strain was cultivated in an erlenmeyer flask under a submerged fermentation system. Culture conditions for the production of a tannase of. Optimization of tannase production by aspergillus niger in solidstate packedbed bioreactor article pdf available in journal of microbiology and biotechnology 219. Biosynthesis and characterization of aspergillus niger. Optimized production of tannase from cashew testa using aspergillus niger mtcc 5898. Agitation speed was found to influence the tannase production and fungal growth of aspergillus niger fetl ft3. Purification, characterization and application of tannase. High performance liquid chromatography and metabolomics. Involvement of physical parameters in medium improvement. Extracellular tannase production was induced when the fungus was cultured in khanna medium with tannic acid as carbon. For tannase production by paecilomyces variotii, temperature range of 4065 c and ph range of 4. Materials and methods microorganism and culture medium the microorganism used for tannase production, a. Rhizophora apiculata bark is a tanninrich waste material obtained from charcoal industry. Tannase activity was also measured in solidstate fermentation.
Among the different temperatures such as 25, 30, 35, 40 and 45c tried, the maximum enzyme production was observed at 35c 48 ugmin. The production of tannase by aspergillus niger gh1 in solidstate fermentation ssf was evaluated. Central composite rotatable design ccrd falling under response surface methodology. Isolation and screening of tannase producing fungi. A method for assay of microbial tannase tannin acyl hydrolase based on the formation of chromogen between gallic acid and rhodanine is reported. Central composite rotatable design ccrd falling under response surface methodology rsm was used. Production of tannase from aspergillus niger under solid.
Aspergillus niger isolate which was proved efficient was used for gallic acid production. Gallic acid production with mouldy polyurethane particles. The optimum temperature for the enzyme activity was found to be 3040c, at which the enzyme activity was. Production of gallic acid using the immobilized cells of aspergillus oryzae has been studied. Bioconversion of naturally occurring tannins into a value. The current study investigated the production of tannase by aspergillus niger cepc 11 mtcc 5898 on an abundantly available waste byproduct cashew testa under solid state a. Pdf optimization of tannase production by aspergillus niger. Culture conditions for the production of a tannase of aspergillus tamarii imi388810. Some of these fungal strains such as aspergillus niger gh1 and psh strain have demonstrated to be excellent enzyme producers. Growth of colonies in plates with tannic acidcontaining medium indicated their ability to synthesize tannase. Opadanje aktivnosti tanaze nakon 20 sati ugoja povezano je s pratecim porastom aktivnosti proteaze. The production of enzymes by bioprocessing is a good alternative to add value to agroindustry residues. Abstract gallic acid 3,4,5 trihydrixybenzoic acid is an organinc substance occuring in many plants either as a free molecule or as part of tannic acid molecule.
Addition of high tannic acid concentrations increased total tannase activity in ssf, while in smf it was decreased. The culture supernatants obtained by filtration through whatman no. Optimisation of fermentation conditions for production of. In this work, we measured tannase in aspergillus niger fermented tea leaves and confirmed the production of fungal tannase during puerh tea fermentation. Production of tannase by rhizopus oryzae and aspergillus foetidus from the powdered fruits of terminalia chebula and caesalpinia digyna has been reported11. Gallic acid production by immobilization of aspergillus orzae. Optimization of gallic acid production from terminalia. Optimized production of tannase from cashew testa using aspergillus niger mtcc 5898 article pdf available in food biotechnology 304.
The ph, incubation period, temperature and glucose concentration optima of tannase production was found at 5. An extracellular tannase was produced from solidstate cultures of aspergillus niger. The authors also performed studies to optimize the process. Although tannase production by aspergillus niger can occur in the absence of tannic acid, this fungus tolerates tannic acid concentrations as high as 20%wv, without having a deleterious effect on both the growth and. Effect of agitation speed on morphological changes in. Economic production of tannase by aspergillus niger van tiegh adopting different fermentation protocols and possible applications romanian biotechnological letters, vol. The enzyme was purified to homogeneity from the cellfree culture broth by preparative isoelectric focusing and by fplc using anionexchange and gelfiltration chromatography. The most active tannase producer was selected and identified as aspergillus niger aumc 4301 at mycological center, faculty of science, assuit university, egypt. Effects of initial substrate tannic acid concentration, incubation time and temperature on tannase production in ssf have been studied. Production cost of enzymes, the major constraint in bioprocessing, may be brought down by multifaceted approaches which include the use of. This study was carried out to optimize the physical parameters for the maximal tannase production by aspergillus niger via solid. Aspergillus niger mtcc 3557 was used for maximum production of tannase enzyme and the factors affecting the production were optimized.
Gallic acid formation from gallotanninsrich agricultural. These properties make the enzyme suitable for pollution control and. There is a significant effect p tannase from fungal sources is reported to be highly active over a wide range of ph and temperature. Pdf optimized production of tannase from cashew testa. In this regard there are no reports on bacterial tannase production using tannin containing any agrobased substrates. Production of tannase using aspergillus niger by submerged. Gallic acid production with mouldy polyurethane particles obtained from solid state culture of aspergillus. The optimum conditions for maximum tannase enzyme production in the submerged fermentation smf using aspergillus niger was. Production, characterization and purification of tannase from. Maximum tannase production occurred in the culture broth containing 12% wv tannic acid and 0.
Bioconversion of naturally occurring tannins into a value added pharmaceutical intermediate gallic acid a new approach. Aspergillus niger holds the generally recognized as safe status from the food and drug authority and is the primary. Tannase production in submerged fermentation by aspergillus sp. The optimum values of parameters obtained through response surface methodology rsm were cashew testa 23%, k. Production of tannase by aspergillus niger ozturk, alev deniz m. Tannase production by aspergillus niger aa20 was studied in submerged smf and solidstate ssf fermentation systems with different tannic acid and glucose concentrations. This fungus was cultivated in 250 ml erlenmeyer flask under submerged fermentation system. Tannase is an enzyme widely used in the food industry, mainly for wine and beer production. This industrial waste was used as solid substrate in the study for the production of tannase and at the same time, help in minimizing the countrys industrial wastes. Sdspage analysis as well as gel localization studies of purified tannase indicated the presence of two enzyme forms, with molecular. Economic production of tannase by aspergillus niger van tiegh. Tannase from fungal sources is reported to be highly active over a wide range of ph and temperature. The optimal agitation speed was at 200 rpm which produced 1.
Optimized production of tannase from cashew testa using. Preliminary culture conditions for aspergillus nigergh1. To demonstrate that fungal tannase excretion is strongly influenced by the culture conditions, tannase production by aspergillus niger gh1 on smc using different bioreactors was evaluated. To study the effect of increasing doses of gamma radiation on tannase production by a. Various physical parameters were studied in order to maximize the tannase production. Tannase production by aspergillus niger 194 was sterilized and inoculated with 2% vv inoculum was added and incubated at 35 c for 36 h in a rotary shaker 200rpm. In this article, aspergillus ochraceus is presented as a new source of tannase with interesting features for biotechnological applications. Lower or higher agitation speeds than 200 rpm produced lower enzyme production and fungal growth. Aspergillus niger is cultured for the industrial production of many substances. The present study explores the natural ability of fungi to utilize tannin as a source of carbon. Aspergillus niger fetl ft3, a local extracellular tannase producer strain that was isolated from one of dumping sites of tanninrich barks of rhizophora apiculata in perak, malaysia. Aspergillus niger fbt1, a local extracellular strain for tannase production, was isolated from soil collected from matang mangrove forest reserve in perak, malaysia. Tannase is also utilized for bioremediation of effluents from tanneries. Optimization of gallic acid production from terminalia chebula 291 from the fig.
The results indicated that the unpasteurized method before fermentation was in favor of the higher enzymic activity than the sterilized method. Aspergillus niger gh1 tannase selfimmobilized during the ssf process. Abstract aspergillus niger is a mould known to be the best tannase producer. Some factors affecting tannase production by aspergillus niger van tieghem hamada a. However, the diversity of fungi stays poorly explored for tannase production.
Pdf original article statistical optimization for tannase. Study is on the solidstate fermentation of tannase with the gallnut by aspergillus niger b0201. The aim of this work was to select strains of aspergillus niger for tannase production. Process parameters influencing tannase production by. Some factors affecting tannase production by aspergillus.
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