Extracellular Tannase Production Research Articles

OPTIMIZATION OF POMEGRANATE PEEL EXTRACTS FOR THE BIOCONVERSION OF THE ELLAGITANNINS TO ELLAGIC ACID USING ASPERGILLUS NIGER, RHIZOPUS ORYZAE AND MIXED CULTURE

Objective: The present study is aimed at optimization of concentration of substrate (pomegranate peel extract) for the production of Ellagic acid using Aspergillus niger and Rhizopus oryzae. Methods: Test organisms were isolated and identified using standard microbiological techniques. Collected pomegranate peels were dried, grained and used as the substrate for solid-state fermentation. Prepared spore suspension of the test organisms was inoculated and incubated at room temperature. At regular intervals of 24 h, samples were drawn and subjected to analysis of reducing sugar, soluble proteins, hydrolysable tannins and production of extracellular tannase using standard methods. Results: The results of the present investigation demonstrated the optimum substrate concentration for the active conversion of ellagitannins to ellagic was 15g of A. niger, 20g for R. oryzae and 15g for mixed culture. Conclusion: From the current work, it was concluded that solid-state bioprocessing of fruit substrates and fruit wastes using fungi has shown to enrich phenolic antioxidants and improve phytochemical consistency.

Kinetic, logistic, box-behnken model and statistical procedure for media optimization of tannase producing bacteria from tannery effluent and its application in Glue and gelatine production for pharma applications

A key enzyme tannase enables the degradation of gallotannins, a type of hydrolysable tannins. A wide variety of microorganisms such as yeasts, fungi and bacteria leads to the production of tannase. Among these the tannase producing bacteria were commonly reported in dissipated water from paper, leather and forestry industries. In this study we will discuss about diverse bacteria that were isolated from the tannery effluent of several industries and its screening, characterization and the production of extracellular tannase. The production of extracellular tannase is highly influenced by Media Optimization and hence wide range of pH and temperature is utilized for the production of tannase and the optimal activity was found to be at 45°C and at a pH 6.5. The placket-Burman Logistic, Kinectic and Box-Behnken models were employed to optimize the media for tannase producing bacteria. In submerged culture (SMC) and solid-state (SSC) the induction and repression pattern of tannase production can be observed and the results presented that SSC has higher potential to minimize catabolite repression. Assessment of the effect of various additives on tannase activity was also done and thus among the various metal salts tested, the enzyme was found to be strongly inhibited byHgCl2 followed by ZnCl2 and MnCl2. Again it was also found that EDTA and ?- mercaptoethanol had inhibitory effect where as the detergents (Tween-20, -60 and -80 as well as SDS) did not affect the activity of the enzyme1. The recovery of glue and gelatin from tannery effluent was also studied and for that a three step hydrolysis process is best suited for maximum recovery of collagen hydrolysate followed by the glue and gelatine production from it. It was also reported in accordance with the antibiotic sensitivity test (Abst) that the bacteria were very sensitive for streptomycin.

Optimization of Tannase Production and Improvement of Nutritional Quality of Two Potential Low-Priced Plant Feedstuffs under Solid State Fermentation by Pichia kudriavzevii Isolated from Fish Gut

An autochthonous tannase yielding yeast strain Pichia kudriavzevii (GU939629), isolated from the gut of an Indian major carp (mrigal), Cirrhinus cirrhosus, has been used for extracellular tannase production and subsequent processing of two plant feedstuffs, Groundnut oil cake (GOC) and Pistia leaves (PL), under solid state fermentation (SSF). Of the two plant materials studied, GOC supported maximum tannase activity (0.82 ± 0.024 U/gds) degrading 94.1% of the initial tannin content, whereas SSF resulted in 0.68 ± 0.02 U/gds tannase activity and 89.1% tannin degradation through the use of PL as substrate. Following SSF for 15 d with optimized culture conditions, analysis of proximate composition revealed that there was significant increase (t-value significant at P < 0.05) in the contents of crude protein, lipid, minerals (Na, K, Ca, Mg, Zn, Fe, Cu, Mn, P), free amino acids and fatty acids; along with reduction in the contents of the other antinutritional factors, for example, crude fiber, phytic acid, and trypsin inhibitor. The results indicate that there is ample scope for further research to appraise potential application of gut microbiota for tannase production, as well as processing of low-cost plant feedstuffs for prospective use as feed ingredients for improved fish protein production.

Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus

Background: Tannases are enzymes that may be used in different industrial sectors as, for example, food and pharmaceutical. They are obtained mainly from microorganisms, as filamentous fungi. However, the diversity of fungi stays poorly explored for tannase production. In this article, Aspergillus ochraceus is presented as a new source of tannase with interesting features for biotechnological applications. Results: Extracellular tannase production was induced when the fungus was cultured in Khanna medium with tannic acid as carbon source. The extracellular tannase was purified 9-fold with 2% recovery and a single band corresponding to 85 kDa was observed in SDS-PAGE. The native apparent molecular mass was estimated as 112 kDa. Optima of temperature and pH were 40ºC and 5.0, respectively. The enzyme was fully stable from 40ºC to 60ºC during 1 hr. The activity was enhanced by Mn 2+ (33-39%) and NH 4 + (15%). The purified tannase hydrolyzed tannic acid and methyl gallate with Km of 0.76 mM and 0.72 mM, respectively, and Vmax of 0.92 U/mg protein and 0.68 U/mg protein, respectively. The analysis of a partial sequence of the tannase encoding gene showed an open read frame of 567 bp and a sequence of 199 amino acids were predicted. TLC analysis revealed the presence of gallic acid as a tannic acid hydrolysis product. Conclusion: The extracellular tannase produced by A. ochraceus showed distinctive characteristics such as monomeric structure and activation by Mn 2+ , suggesting a new kind of fungal tannases with biotechnological potential. Further, it was the first time that a partial gene sequence for A. ochraceus tannase was described.

Media optimization for extra cellular tannase production by Klebsiella pneumoniae MTCC 7162 using response surface methodology

Klebsiella pneumoniae MTCC 7162, isolated from tannery effluent was used for the production of extracellular tannase. Central composite design (for nutritional parameters) and Box Behnken design (for physicochemical parameters) were used to arrive at conditions for maximal tannase production. Box Behnken design was statistically significant and indicated that pH is a significant factor for tannase production. A 6.5 fold increase of extra cellular tannase (22.7 U / ml) production was observed at the optimized conditions. The yield of extra cellular tannase obtained in this study, from a bacterial source, is the highest compared to that of the earlier reports. Key words: Klebsiella pneumoniae, extra cellular tannase, central composite design, Box Behnken design, response surface methodology.

Production of tannase from wood-degrading fungus using as substrate plant residues: purification and characterization

In the present study Lenzites elegans, Schizophyllum commune, Ganoderma applanatum and Pycnoporus sanguineus (wood-degrading fungi) were assayed for their tannase producing potential in culture media containing plant residues or/and tannic acid as carbon source. Aspergillus niger was used as positive control for tannase production. We also carried out the isolation, purification and characterization of the enzyme from the fungi selected as the major productor. The highest fungal growth was observed in A. niger and L. elegans in the media containing tannic acid + glucose + plant residues (Fabiana densa). A. niger and L. elegans reached the highest extracellular tannase production in a medium containing tannic acid + F. densa and in a medium supplemented with glucose + tannic acid + F. densa. The produced enzyme by L. elegans was purified by DEAE-Sepharose. Km value was 5.5 mM and relative molecular mass was about 163,000. Tannase was stable at a pH range 3.0–6.0 and its optimum pH was 5.5. The enzyme showed an optimum temperature of 60°C and was stable between 40 and 60°C. This paper is the first communication of tannase production by wood-degrading fungi. Fermentation technology to produce tannase using plant residues and xylophagous fungi could be very important in order to take advantage of plant industrial waste.

Production and partial purification of extracellular tannase by Klebsiella pneumoniae MTCC 7162 isolated from tannery effluent

Diverse bacteria belonging to various taxa were isolated from tannery effluent of leather industries located at Ranipet, India and were screened for the production of extracellular tannase. One of the strains identified as Klebsiella pneumoniae MTCC 7162 was found to produce tannase (3.4 U/ ml) at pH 6.0, 37°C and 100 rpm. Use of individual carbon and inorganic nitrogen sources resulted in lower tannase production. However, a combination of urea and corn steep liquor extract yielded marginal increase in tannase production (3.9 U/ ml). End-product repression was also studied with inclusion of gallic acid to the growth medium. Enrichment with various additives of metal ions and detergents resulted in inhibition of tannase production. The enzyme was partially purified using ammonium sulfate precipitation followed by the use of DEAE-cellulose. SDS-PAGE analysis indicated that the molecular weight of the protein to be 46.5 kDa. The enzyme was found to be active in a wide range of pH and temperature with an optimal activity at pH 5.5 and 40°C.

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% reduction of tannic acid content in case of jamun wine, 43.59% reduction in case of grape wine and 74% reduction in the tea extract after 3 h at 35°C.

Optimisation of extracellular tannase production from Paecilomyces variotii

The tannase production by Paecilomyces variotii was confirmed by high performance thin layer chromatography (HPTLC), and substrate specificity of the tannase was determined by zymogram analysis in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS–PAGE). A clear band of activity observed after electrophoresis of culture filtrate in non-denaturing gels indicated the production of extracellular tannase by P. varoitii. HPTLC analysis revealed that gallic acid was the enzymatic degradation product of tannic acid during the fermentation process. The optimum condition for tannase production was at 72 h of incubation in shaking condition and addition of 1.5% tannic acid, 1% glucose and 0.2% sodium nitrate at temperature of 35°C and pH of 5–7. The production of extracellular tannase from Paecilomyces variotii was investigated under optimized conditions in solid-state fermentation (SSF), submerged fermentation (SmF) and liquid surface fermentation (LSF) processes. The maximum extracellular tannase production was obtained within 60 h of incubation under SSF followed by SmF and LSF.

Extracellular tannase production by endophytic Hyalopus sp.

Extracellular tannase production by endophytic Hyalopus sp.

Studies on the extracellular tannase from newly isolated Bacillus licheniformis KBR 6.

A tannase producing bacterial strain KBR 6 has been isolated from lateritic soil and identified as Bacillus licheniformis. It is capable of producing tannase in the medium containing only tannic acid. The rapid degradation of tannic acid and production of extracellular tannase was observed in three different media containing tannic acid (M1), tannic acid + basal salt (M2) and tannic acid + basal salt + glucose (M3). Maximum enzyme production and growth of the organism was obtained at 18-21 h and 30-36 h, respectively. The increased order of enzyme production in relation to different media is as per the following sequence, M3 > M2 > M1. The maximum growth and enzyme production was observed at pH 5.0. The pH and temperature optima of the enzyme activity were found to be at 5.75 and 60 degrees C respectively. Paper chromatographic analysis indicates that gallic acid is the enzymatic degradative product of tannic acid.