Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable <i>Bacillus amyloliquefaciens</i> BA-DES4

ZHANG Zhi; WEN Dongzhuo; FENG Lirong; ZHANG Shenglong; YANG Kexin; ZHANG Xiaotong

doi:10.13386/j.issn1002-0306.2022070337

Volume 44 Issue 11

May 2023

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Science and Technology of Food Industry > 2023 > 44(11): 136-143. > DOI: 10.13386/j.issn1002-0306.2022070337

ZHANG Zhi, WEN Dongzhuo, FENG Lirong, et al. Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4[J]. Science and Technology of Food Industry, 2023, 44(11): 136−143. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022070337.

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Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4

1.
School of Forestry, Northeast Forestry University, Harbin 150040, China
2.
Heilongjiang Guohong Energy Conservation and Environmental Protection Co., Ltd., Harbin 150040, China

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Received Date: August 01, 2022
Available Online: April 01, 2023

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Abstract

Objects: This research was performed based on taking one Bacillus amyloliquefaciens BA-DES4 producing the cellulose as the material, and the enzymatic property of its cellulose was researched upon the purification. Methods: This research applied the ammonium sulfate fractional precipitation and SephadexG-75 gel filtration chromatography method to carry out the separation and purification to the cellulose it produced. Determined its molecular weight by means of polyacrylamide gel electrophoresis (SDS-PAGE), and carried out the research of enzymatic property to enzyme solution of cellulase upon purification. Results: The cellulase system components could be obtained in zymotic fluid upon separation and purification (endoglucanase), and its specific activity was 51.08 U/mg through carrying out the measurement of enzyme activity to purified electrophoresis endoglucanase. It was identified that its molecular weight was 22.4 kDa. The primary research of enzymatic property indicated that the optimal reaction temperature and optimal pH for this enzyme were 65 ℃ and 6.0 respectively with a high stability when pH was 5.0～7.0 and temperature was 55～65 ℃. The activation of Mn²⁺ to cellulase activities was significant with the highest inhibiting effect of Cu²⁺. Conclusion: This bacterial strain can be used as the potential strain for producing the endoglucanase with the enzymatic activities of endoglucanase can be promoted under the effects of Mn²⁺, Fe²⁺. Meanwhile, it can work under the high temperature and acid environment along with participating in the cellulose of high effective degradation and high-temperature acid environment so as to improve the production efficiency. The glucose to be decomposed can be used for fermentation with a potential of producing the fermented wine by applying the high-temperature Daqu, which has laid a foundation for further studying this cellulase.
- Bacillus amyloliquefaciens,
- cellulase,
- purification,
- enzymatic property

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