Maximum Enzyme Production Research Articles

Optimization and strain improvement by mutation for enhanced cellulase production by Bacillus sp. (MTCC10046) isolated from cow dung

The goal was to investigate the cellulase enzyme production ability of bacterial strain C1 isolated from cow dung and identified as Bacillus sp. on the basis of 16 S rDNA sequence homology. The effects of different carbon sources like Carboxymethyl cellulose (CMC), avicel, starch, maltose, sucrose, glucose, fructose, galactose and lactose on cellulase production at varying environmental parameters of incubation period (2–10days), temperature (35–55°C), and pH (6.0–8.5) were examined. The CMC was the best carbon source for cellulase production followed by lactose in this bacterial strain. The maximum enzyme production was achieved at a temperature of 50°C by Bacillus sp. with pH of 7.0 on the 8th day of growth. The nitrogen source NH4NO3 at 0.175% was optimum for cellulase production by this bacterium. A putative mutant (C1M26) was screened from wild C1 strain after mutagenesis with N-methyl-N′-nitro-N-nitrosoguanidine (NTG) as a mutagenic agent. The mutant C1M26 produced a larger amount of cellulase in comparison to wild type C1 strain.

Partial characterization of a crude cold-active lipase from Rhodococcus cercidiphylli BZ22.

Cold-active lipase production by the psychrophilic strain Rhodococcus cercidiphylli BZ22 isolated from hydrocarbon-contaminated alpine soil was investigated. Depending on the medium composition, high cell densities were observed at a temperature range of 1-10 °C in Luria-Bertani (LB) broth or 1-30 °C in Reasoner's 2A (R2A). Maximum enzyme production was achieved at a cultivation temperature of 1-10 °C in LB medium. About 70-80% of the secreted enzyme was bound to the cell and was highly active as a cell-immobilized lipase which exhibited good reusability; more than 60% of the initial lipase activity was retained after five-fold reuse. The properties of the lipase produced by the investigated strain were compared with those of a mesophilic porcine pancreatic lipase (PPL). The thermal stability of the cell-immobilized bacterial lipase was higher than that of the extracellular enzyme. Highest activity was detected at 30 °C for the cell-immobilized enzyme and for PPL, while the extracellular enzyme displayed highest activity at 10-20 °C. The bacterial lipase hydrolyzed p-nitrophenyl (p-NP) esters with different acyl chain lengths (C2-C18). The highest hydrolytic activity was obtained with p-NP-butyrate (C4) as substrate, while the highest substrate affinity was obtained with p-NP-dodecanoate (C12) as substrate, indicating a clear preference of the enzyme for medium acyl chain lengths.

Streptokinase production from Streptococcus dysgalactiae subsp. equisimilis SK-6 in the presence of surfactants, growth factors and trace elements.

Streptokinase is an extracellular protein secreted by various strains of streptococci and is used clinically as an intravenous thrombolytic agent for the treatment of acute myocardial infarction. It is well established that the fibrinolytic activity of streptokinase originates in its ability to activate plasminogen. The present investigation was carried out to determine the extent of streptokinase production by Streptococcus dysgalactiae subsp. equisimilis SK-6, in the presence of surfactants, growth factors, trace elements and under the influence of different physical parameters. Mineral salt medium was supplemented with different surfactants, growth factors and trace elements. Effects of incubation period and inoculum volume were also studied. Results indicated that the streptokinase yield was higher in the presence of non-ionic surfactants, where Tween 80 supported maximum enzyme production (0.178 U/ml). Growth factors such as glycine and thiamine supplementation resulted in better enzyme production. Trace elements in the form of magnesium sulphate and ferrous sulphate when added in lesser quantity aided higher streptokinase synthesis. Enzyme production was severely affected in the presence of higher concentrations of these inorganic salts. A constant decrease in the enzyme production was observed beyond 48 h of incubation. Among the different inoculum sizes used, 1 % v/v inoculum facilitated highest streptokinase production (0.360 U/ml). The streptokinase production ability of S. dysgalactiae subsp. equisimilis SK-6 offers its potential industrial application for the manufacture of streptokinase.

Optimization of gelatinolytic enzyme production by B. amyloliquefaciens sp. H11 through Plackett–Burman design and response surface methodology

Bacillus amyloliquefaciens H11 has been proven as a potential producer of extracellular protease with capacity of hydrolyzing gelatin. Therefore, the cultivation conditions for the enhanced production of gelatinolytic enzyme from a newly isolated B. amyloliquefaciens H11 was investigated using Plackett- Burman design and response surface methodology. Three significant variables (agitation speed, cultivation time and fish gelatin concentration) were selected for optimization. Increase in speed of agitation and fish gelatin concentration markedly increased the production of gelatinolytic enzyme. Gelatin concentration and cultivation time showed significant interaction and both variables played the important role in enzyme pro- duction. The maximal gelatinolytic enzyme production in the basal medium was 2,801 U/mL under the following optimal condition: agitation speed of 234 rpm, 8.36 g/L of fish gelatin and 31 h of cultivation. The predicted model fitted well with the experimental results (2,734 ± 101 U/mL). 14-fold increase in yield was achieved, compared with the basal condition (212 U/mL). Thus, cultivation of B. amyloliquefaciens H11 under the optimal condition could enhance the production of gelatinolytic enzyme effectively.

Scale up of production in a bioreactor of a halotolerant protease from moderately halophilic Bacillus sp. isolated from soil

Studies were conducted on the production of protease by moderately halophilic Bacillus sp. on agro-industrial waste materials. The bacterium could efficiently use many agro wastes as substrates but wheat bran supported maximum enzyme production. To ascertain the performance of the process in shake flasks and lab scale bioreactor, experiments were conducted to analyse protease activity utilizing wheat bran as cost effective substrate. The studies unveiled that pH 7.0, temperature 30°C and static conditions were optimal for enzyme production in flask level fermentation. In scale-up fermentation, at optimal pH and temperature, agitation rate of 50 rpm was best for protease production. The enzymatic nature was studied in 10% SDS gels with BSA (2.5 mg/mL) as substrate and banding pattern was compared with undigested BSA as control. The endoprotease nature and the kinetics of protease activity were confirmed. The enzyme retained 37% of its activity even at 5 M NaCl concentration. The proteolytic activity was also confirmed by casein zymogram analysis. The fermentation medium containing inexpensive substrates, physical conditions and ability of Bacillus sp. to exhibit protease activity on a large scale could collectively be useful for commercial production.

Process optimization and production kinetics for cellulase production by Trichoderma viride VKF3

Microbial cellulases are the enzymes widely studied due to their enormous applications in biochemical industry. Among 12 fungal isolates isolated from mangrove plant debris and soil sample collected from Valanthakad Mangroves, Kerala, India, 3 of them were found to exhibit cellulolytic activity. Among them, the most potent isolate which exhibited maximum cellulolytic activity was identified as Trichoderma viride VKF3 [Gene bank accession number- JX683684.1] based on colony morphology, microscopic observation and molecular centeracterization using D1/D2 region amplification. The isolate T. viride VKF3 was found to be non-phytopathogenic against the selected plants. Neighbour joining tree depicted its least divergence rate from the root taxon HM466686.1. T. viride VKF3 was grown under dynamic carbon, nitrogen sources, pH and temperature of the medium to draw out the optimum conditions for cellulase production. Protein stability kinetics and biomass production was also studied upto 11th day of incubation. It was evident from the study, that dextrose and beef extract could be used as major carbon and nitrogen sources in submerged fermentation at pH 9.0 and incubation temperature of 25°C to get maximum CMCase yield. Optimum enzyme recovery period was identified between 5th to 9th days of incubation beyond which the enzyme activity was reduced. By comparing two fermentation methods, submerged fermentation was found to be the best for maximum enzyme production. But utilization of substrates like sugarcane bagasse and cassava starch waste in the SSF offers a better scope in biodegradation of solid waste contributing to solid waste management.

<b>Critical process parameters optimization for hyperthermostable β amylase production by <i>Bacillus subtilis</i> DJ5 using response surface methodology </b> - doi: 10.4025/actascibiolsci.v36i1.17427

The combined effects of significant physical and chemical factors affect hyperthermostable β amylase production under submerged fermentation by Bacillus subtilis DJ5. The above was studied using the experimental design and response surface methodology. A 23 full-factorial central composite design was chosen to analyze interactions among three factors i.e. substrate concentration, medium pH and incubation temperature. The experimental data were fitted into a polynomial model for the yield of enzyme and an optimum level was arrived at with optimized conditions. Solving the coded values using Excel equation function indicated that maximum enzyme production is possible at a substrate concentration of 7.07 mg mL-1, pH 6.622 and temperature of 35.435°C. Such prediction was validated with practical experiments in which, at the prescribed condition maximum yield of 15.62 U mg-1, nearly 1.5 fold higher than non-optimized condition was observed.

Enhancing the efficiency of the bioagent Bacillus subtilis JF419701 against soil-borne phytopathogens by increasing the productivity of fungal cell wall degrading enzymes

This study was devoted to increasing the production of fungal cell wall degrading enzymes by Bacillus subtilis JF419701 to enhance its efficiency in the biological control process. In dual culture, B. subtilis JF419701 showed the highest antagonistic effect of the 256 bacterial strains tested against six soil-borne pathogens, Alternaria alternata, Exserohilum rostratum, Fusarium oxysporum, Macrophomina phaseolina, Pythium ultimum and Rhizoctonia solani. The production potentiality of the enzymes α-1,3-glucanase, β-1,3-glucanase, chitinase and protease by B. subtilis JF419701 was studied in vitro. Results proved that the maximum production of enzymes by this bacterium was achieved after a two-day incubation period at a slightly alkaline pH (8). The addition of colloidal chitin or S-glucan to the growth media enhanced the production of all the enzymes except protease, which was stimulated by casein. This study therefore recommends that to obtain an efficient and strong bioagent culture of B. subtilis JF419701, it is necessary to grow this micro-organism on a specific medium containing either chitin or its derivatives at pH 8 for two days.

Enhanced production of cellulose degrading CMCase by newly isolated strain of Aspergillus versicolor

Production of CMCase from newly isolated strain of Aspergillus versicolor was studied by optimizing different physico-chemical parameters. Cellulolytic potential of the isolate was confirmed by the simple agar plate method and then subjected to variation of a single parameter at a time in CMC containing medium. Maximum enzyme production was obtained at 30°C in 120h when medium was supplemented with 0.5gm% carboxymethyl cellulose, 0.2gm% Tween 80, 0.075gm% peptone, 0.050gm% CaCl2, 1.5gm% NaNO3 and 0.2gm% KH2PO4 at pH 4.0. Current study holds great potential as the organism reported here could easily be used for the production of cellulase using renewable biomass.

Utilization of Agro-Industrial Waste Residues for the Production of Amylase from Aspergillus oryzae IIB-6

Aims: The aim of this study was to evaluate different solid substrates and moistening agents for the production of an extracellular 1, 4-α-D-glucan glucohydrolase from a newly isolated Aspergillus oryzae IIB-6 by solid state fermentation and optimization of culture conditions for the maximal production of enzyme. Study Design: Six different agro-industrial waste residues (rice straw, rice bran, corn flakes, wheat bran, wheat flakes, and grinded wheat kernel) were procured from the local market. These substrates (10 g) were moistened (1:1) with different moistening agents (distilled water, tap water, mineral salts solution (FeSO4.7H2O 0.02, MgSO4.7H2O 1.0, (NH4)2SO4 4.0, KH2PO4 0.6, K2HPO4 1.4 mg/gds at pH 5), 0.1 N HCl, sodium acetate buffer (pH 5.5), sodium phosphate buffer pH 7.5) and screened for the production of 1,4-αD-glucan glucohydrolase for 96 hours in static cultures. The substrate and moistening agent that gave maximum enzyme production were selected and their fermentation conditions were further optimized. Place and Duration of Study: Institute of Industrial Biotechnology (IIB), GC University Lahore, Kachery Road Lahore, Pakistan, between February 2011 and March 2012. Methodology: The levels of selected solid substrate, moistening agent and fermentation conditions such as pH, temperature, time of incubation, inoculum size etc. were optimized Original Research Article British Biotechnology Journal, 4(4): 350-365, 2014 351 by one variable at a time method. Results: Wheat bran at the level of 5 g and mineral salts solution containing FeSO4.7H2O 0.016, MgSO4.7H2O 0.8, (NH4)2SO4 3.5, KH2PO4 0.48, K2HPO4 1.12 mg/gds gave relatively best enzyme production. The maximum enzyme activity 7800 U/gds (407 Umg protein) were achieved when wheat bran with 80% moisture content was incubated at 30°C, pH 5, after inoculating with 10% spore suspension (1.2×10 CFU/gds) for 72 h. Conclusion: Aspergillus oryzae IIB-6 was a good producer of 1,4-α-D-glucan glucohydrolase in wheat bran medium containing mineral salts as an additional trace elements so that it can be used for biotechnological purposes.

Production and characterization of cellulolytic enzymes by isolated<br>Klebsiella sp. PRW-1 using agricultural waste biomass

The efficient cellulolytic microorganism was isolated from soil samples collected from Shivaji University campus. Biochemical test and phylogenetic analysis of isolated culture identified as Klebsiella sp. PRW-1. The isolated culture could utilize pure cellulosic substrates (carboxymethylcellulose (CMC) and avicel) and different agricultural wastes like sugarcane bagasse, sugarcane barbojo, sorghum husks, grass powder, corn straw and paddy straw by producing a large amount of endoglucanase, exoglucanase, β-glucosidase, filter paperase (FPU), xylanase and glucoamylase. The reducing sugar production was found higher in the presence of grass powder and sugarcane barbojo. Effects of different physic-chemical parameters to achieve maximum cellulolytic enzymes production were systematically investigated. The effects of supplementation of different metals additives as well as the stability in the presence of higher temperature, pH and commercial detergents, on cellulolytic enzymes were also studied. The foregoing result increases the applicability of the strain for the utilization and bioconversion of lignocellulosic biomass that could be used for bioenergy production.

OPTIMIZATION OF CULTURAL PARAMETERS FOR CELLULASE ENZYME PRODUCTION FROM FUNGI

The objective of this study is to reduce the production cost of cellulase by optimizing the production medium and using an alternative carbon source such as agricultural solid waste residue. Present studies describe the optimization of process parameters for the production of cellulases by different fungal species. The fermentation experiments were carried out in shake flasks. Maximum production of cellulases (0.63 U/ml) was observed after a fermentation period of 72 hrs at an temperature of 40° C. Initial pH of the culture medium was also optimized and a pH of 5 was found to support maximum growth and enzyme production (0.9 U/ml) by A. niger. Different inorganic nitrogen sources and carbon sources were evaluated for the production of cellulases and ammonium sulphate in case of nitrogen and glucose among carbon sources was found to be the best. Agrobased coconut cake waste gave the best production of cellulases. Cellulase production from A. niger and Penicillium sp can be an advantage as the enzyme production rate is normally higher as compared to other fungi.

Production of an extracellular lipase by Candida utilis NRRL-Y-900 using agro-industrial by-products

Aim:To screen various yeast cultures and agro-industrial by-products and optimization of fermentation conditions for the microbial production of extracellular lipase under solid state fermentation technique. Material and Methods: Various yeast cultures including Candida lipolytica NRRL-Y-1095, Candida utilis NRRL-Y-900, Candida tropicalis NRRL-Y-1552, Saccharomyces cerevisiae IIB-1 were screened by culturing on agro-industrial by-products under batch culture using solid state fermentation in 250 mL Erlenmeyer flasks. The medium was supplemented with various nitrogen sources (both organic and inorganic) and metal ions. Results: Candida utilis NRRL-Y-900 showed the highest enzyme production on soybean meal. Various particle sizes of substrate and moistening agents were also optimized for the maximum lipase synthesis. The optimum temperature and pH for the accumulation of enz- yme by Candida utilis NRRL-Y-900 was 30oC and 6.5, respectively. The fermentation time of 60h was suitable for the maximum enzyme production by using 7.5% inoculum of 24h old yeast culture. The optimal medium composition consisted of 2% (w/v) meat extract, 0.4% (w/v) ammonium sulphate and 5mM Fe+2. The maximum extracellular lipase production was 3.96±0.09 U. Conclusion: The results obtained during the study are significant for, to our knowledge, it is the first report regarding the utilization of soybean meal by C. utilis NRRL-Y-900 to accu- mulate lipase under optimized conditions and solid state fermentation.

A novel promising strain of Trichoderma evansii (WF-3) for extracellular α-galactosidase production by utilizing different carbon sources under optimized culture conditions.

A potential fungal strain of Trichoderma sp. (WF-3) was isolated and selected for the production of α-galactosidase. Optimum conditions for mycelial growth and enzyme induction were determined. Basal media selected for the growth of fungal isolate containing different carbon sources like guar gum (GG), soya bean meal (SM), and wheat straw (WS) and combinations of these carbon substrates with basic sugars like galactose and sucrose were used to monitor their effects on α-galactosidase production. The results of this study indicated that galactose and sucrose enhanced the enzyme activity in guar gum (GG) and wheat straw (WS). Maximum α-galactosidase production (213.63 UmL−1) was obtained when the basic medium containing GG is supplemented with galactose (5 mg/mL). However, the presence of galactose and sucrose alone in the growth media shows no effect. Soya meal alone was able to support T. evansii to produce maximum enzyme activity (170.36 UmL−1). The incubation time, temperature, and pH for the maximum enzyme synthesis were found to be 120 h (5 days), 28°C, and 4.5–5.5, respectively. All the carbon sources tested exhibited maximum enzyme production at 10 mg/mL concentration. Among the metal ions tested, Hg was found to be the strongest inhibitor of the enzyme. Among the chelators, EDTA acted as stronger inhibitor than succinic acid.

Aspergillus fumigatusNITDGPKA3 Provides for Increased Cellulase Production

A cellulolytic fungal strain,Aspergillus fumigatusNITDGPKA3, was isolated from straw retting ground. Cellulase and xylanase production byA. fumigatusNITDGPKA3 in submerged fermentation of rice straw was studied. The culture conditions for maximum enzyme production were found to be initial pH 4, 1% substrate concentration, temperature 30°C, incubation time 5 days, 0.2% tryptone as nitrogen source, and inoculum volumes 7% v/v (for cellulase) and 5% v/v (for xylanase). Addition of Tween 80 in fermentation broth improved xylanase production (193.58 IU/ml) much more compared to cellulase production (6.53 IU/ml). Xylanase activity found in the culture broth was approximately 50% higher compared to most of the reported data. The crude enzyme was further applied for reducing sugar production from alkali pretreated rice straw, where a dosage of 40 IU/g CMCase produced 0.522 g reducing sugar/g dry substrate after 36 hours which was higher than that in the reported literature. The high concentration of reducing sugar yield was most probably due to the extraordinarily high titer ofβ-glucosidase (80.1 IU/ml) found in the crude enzyme. The crude enzymes secreted byAspergillus fumigatusNITDGPKA3 efficiently hydrolyzed alkali pretreated rice straw suggesting thatAspergillus fumigatusNITDGPKA3 is a robust microorganism.

Evaluation of orange peel, an industrial waste, for the production of Aspergillus sojae polygalacturonase considering both morphology and rheology effects

Orange peel is an agroindustrial waste rich in pectin and known to be an inducer for pectinase production. The use of this low-cost substrate for the production of an industrially important enzyme, polygalacturonase (PG), can be an alternative way to turn this waste into a value-added product, contributing to the reduction of environmental waste disposal problems. Enzyme productions by fungal microorganisms are affected by environmental and nutritional factors, demanding the determination of optimum conditions for maximum enzyme production with the desired fungal morphology and broth rheology. Therefore, complex and additional carbon sources were optimized with respect to PG production by Aspergillus sojae using statistical approaches. Effect of pH, another significant parameter affecting the rheology and morphology of the strain, was investigated in the serial bioreactor system using the optimized medium composition. Highest PG enzyme yield and productivity together with the maximum PG enzyme production (93.48 U/mL) were obtained under uncontrolled pH conditions. Under these conditions, morphologically, pellet sizes exhibited a normal distribution ranging between 0.5-1.0 mm and 1.0-1.5 mm, and rheological measurements revealed that fermentation broths showed non-Newtonian flow. The low pH trend observed during the course of the fermentation was another important positive outcome for industrial fermentations, prone to contamination problems.

English

To use oil palm empty fruit bunch (EFB) for cellulase production, a novel fungus was isolated from moistened EFB. This fungus was classified as Penicillium sp. by sequence analysis of the internal transcribing space and it was named Penicillium sp. GDX01. The Penicillium sp. strain secreted cellulases under solid-state fermentation of EFB. The fermentation conditions were optimized for maximal enzyme production. Of the different substrates tested, both EFB and rice straw gave the maximum production of filter paper activity (FPase). Five percent yeast extract and 40-50% initial moisture content were found to be optimal for enzyme production. In addition, the pretreatment of EFB with NaOH before fermentation inhibited the cellulase production of Penicillium sp. GDX01. Saccharification of pretreated EFB by cellulases from Penicillium sp. GDX01 resulted in a more than 80% release of glucose during a 72 h incubation, which is a better result than when using Celluclast 1.5L without the addition of β-glucosidase. Our results show that the cellulases produced by Penicillium sp. GDX01 are more efficient at the saccharification of EFB than Celluclast 1.5 L.   Key words: Cellulase, oil palm empty fruit bunch, solid-state fermentation, Penicillium.

Production, Partial Purification and Kinetic Characterization of Dextransucrase from Leuconostoc mesenteroides in Relation to Dextran Problem in Sugarcane

Dextransucrase was produced from Leuconostoc mesenteroides strain MTCC 107. Maximum enzyme production was observed after 60 h of incubation, with drop in pH from 7.0 at 0 h to 5.5 at 60 h. The crude enzyme was subjected to ammonium sulphate fractionation and DEAE-cellulose anion-exchange chromatography. Three isoforms of dextransucrase viz. DS-I, DS-II and DS-III, each with optimum pH 5.5 and optimum temperature 45 °C were identified. Isoforms DS-I and DS-III were found to be thermostable between 10 and 60 °C while isoform DS-II was thermostable between 10 and 50 °C. The Km values (for sucrose as substrate) of the isoforms DS-I, DS-II and DS-III at their optimum pH and temperature were 206.34, 492.45 and 108.00 mM, respectively. Corresponding Vmax values were 428.08, 505.58 and 410.85 μmol reducing sugars produced/min/mg protein, respectively. The pK a values of ionizing groups obtained by using Dixon and Marangoni methods suggested the participation of Asp/Glu residues and His residues in both substrate binding and catalysis. Lineweaver Burk plots obtained by varying concentrations of dextran (as substrate) at fixed sucrose concentration were found to be biphasic for all the three dextransucrase isoforms, suggesting negative cooperativity. Both the transferase and sucrase activities of the dextransucrase isoforms were markedly inhibited by sodium lauryl sulphate, barium chloride, EDTA, potassium chloride, manganese chloride and sodium metasilicate. These metal salts/chemicals when exogenously added to sugarcane juice effectively reduced the dextran formation during storage.

Production of Tannase using Aspergillus niger by Submerged Fermentation

A study on optimization of submerged fermentative production of tannase by Aspergillus niger was carried out. Aspergillus niger (MTCC 3557) was used for maximum production of Tannase enzyme and the factors affecting the production were optimized. The optimum conditions for maximum tannase enzyme production in the Submerged Fermentation (SmF) using Aspergillus niger was obtained with an initial tannic acid concentration of 3% (w/v) at 35°C, Initial pH of 5.5 and a fermentation period of 96h.The substrates Betalnut husk and Plantain flower gave a maximum tannase production of 19.3 U/ml and 17.89 U/ml respectively.

A New Native Source of Tannase Producer, Penicillium sp. EZ-ZH190: Characterization of the Enzyme

Background: Tannase can be obtained from the various sources for example tannin rich plants; however microbial sources are preferred for industrial production. In microbial sources, the Aspergillus and Penicillium genus and lactic acid bacteria mostly produce tannase. However, it has been identified that this enzyme is produced by many fungi and bacteria, but researches are continuing to find new species. Objectives: The aim of this study was to isolate a tannase-producing fungi from moldy tea leaves and to study some properties of its enzyme. Materials and Methods: The present study was done via two steps. At first, industrially important tannaseproducing fungi were isolated from moldy tea leaves using the simple agar plate method followed by the screening of organisms capable of producing tannase using the enrichment culture technique in modified Czapek Dox’s agar. Finally, tannase obtained from the best isolate was partially purified and characterized. Results: Tannase produced by Penicillium sp. EZ-ZH190 isolated from moldy tea leaves was partially purified and characterized. Maximum enzyme production (4.33 U.mL-1) was recorded after 96 hours of incubation at 30 °C in submerged culture (100 rpm) utilizing 1% (w/v) tannic acid as a sole carbon source. This tannase exhibited optimum activity at 35 °C and at pH of 5.5, and showed nearly 50% of its maximal activity at 50 °C. In the present study, tannase from Penicillium sp. EZ-ZH190 had KM and Vmax values of 1.24 mM and 17.09 U.mL-1, respectively, and showed more than 50% stability at salt (NaCl) concentration of 1 M for 24 hours. Conclusions: Tannase productivity of Penicillium sp. EZ-ZH190 (0.045 U.mL-1.h-1) is comparable with the maximum tannase productivity in the reported literatures, and the biochemical characteristics showed by Penicillium sp. EZ-ZH190 tannase are considered favorable for tannin biodegradation in the industry. So, we concluded that Penicillium sp. EZ-ZH190 is a good strain for use in the efficient production of tannase.