一株高产内切纤维素酶贝莱斯芽孢杆菌的产酶条件优化及酶学性质分析

doi:10.16431/j.cnki.1671-7236.2019.05.012

Abstract

Abstract:

This study was aimed to test the optimization and characterization of high-yield cellulase produced by an endophytic Bacillus velezensis (B.velezensis) 157 from Eucommia bark,which provided a reference for exploring the relevant characteristics of B.velezensis 157 CMCase.Single-factor experiment was designed to optimize fermentation conditions such as pH,inoculation amount,temperature,fermentation time,carbon source and nitrogen source,and to explore the effects of temperature,pH,substrate specificity,metal ions and surfactants on the enzymatic properties of CMCase.The optimized conditions of CMCase production by B.velezensis 157 were as follows:Maize straw as carbon source (30 g/L),soybean meal as nitrogen source (30 g/L),3% inoculation,and pH 7.0 under the condition of 37℃ for 48 h.After the optimization,the CMCase activity of B.velezensis 157 was up to (5.14±0.18) U/mL.The CMCase properties analysis of B.velezensis 157 found that the optimal temperature of enzyme reaction was 60℃,it could still maintain above 80% cellulase activity when temperature was from 40 to 50℃;With the increase of temperature,the relative residual enzyme activity decreased,and it decreased to 50% when the temperature was higher than 55℃.The optimal pH of cellulase was 5.0,and it could still maintain over 90% cellulase activity in a pH range of 5.0 to 10.0;The pH had little effect on the relative residual enzyme activity with the extension of the action time.The cellulase from B.velezensis 157 belonged to typical endoglueanases.Na⁺,Mg²⁺,Ca²⁺ could increase the activity of B.velezensis 157 CMCase,Co²⁺,Hg²⁺,Fe²⁺,Cu²⁺ and SDS inhibited the activity of CMCase;The surfactant Tween-80,Tween-20 and Triton X-100 had no effect on CMCase activity.In this study,the enzyme production conditions and enzymatic properties of B.velezensis 157 CMCase were optimized,and it was concluded that B.velezensis 157 had a high yield of CMCase and tolerated a wide range of pH,which had certain application value in feed additive,detergent and paper industry.

Key words: Bacillus velezensis 157; endocellulase; enzyme production; conditions optimization; enzymatic properties

CLC Number:

Q936

CHEN Long, WU Xingli, LI Lijia, WEI Bingdong, YU Wei, YAN Xiaogang, GU Wei, XU Haiyan, LIU Chen, LIU Hongliang, ZHANG Fangyu. Optimization of Enzyme Production Conditions and Analysis of Enzymatic Properties of a High-yield Endocellulolytic Enzyme Bacillus velezensis[J]. , 2019, 46(5): 1353-1361.

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Optimization of Enzyme Production Conditions and Analysis of Enzymatic Properties of a High-yield Endocellulolytic Enzyme Bacillus velezensis

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