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

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 Seph...

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Main Authors: Zhi ZHANG, Dongzhuo WEN, Lirong FENG, Shenglong ZHANG, Kexin YANG, Xiaotong ZHANG
Format: Article
Language:zho
Published: The editorial department of Science and Technology of Food Industry 2023-06-01
Series:Shipin gongye ke-ji
Subjects:
Online Access:http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022070337
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author Zhi ZHANG
Dongzhuo WEN
Lirong FENG
Shenglong ZHANG
Kexin YANG
Xiaotong ZHANG
author_facet Zhi ZHANG
Dongzhuo WEN
Lirong FENG
Shenglong ZHANG
Kexin YANG
Xiaotong ZHANG
author_sort Zhi ZHANG
collection DOAJ
description 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.
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spelling doaj.art-427613c070f541d1a34f6a7c3a0963552023-08-24T06:01:18ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062023-06-01441113614310.13386/j.issn1002-0306.20220703372022070337-11Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4Zhi ZHANG0Dongzhuo WEN1Lirong FENG2Shenglong ZHANG3Kexin YANG4Xiaotong ZHANG5School of Forestry, Northeast Forestry University, Harbin 150040, ChinaSchool of Forestry, Northeast Forestry University, Harbin 150040, ChinaHeilongjiang Guohong Energy Conservation and Environmental Protection Co., Ltd., Harbin 150040, ChinaHeilongjiang Guohong Energy Conservation and Environmental Protection Co., Ltd., Harbin 150040, ChinaSchool of Forestry, Northeast Forestry University, Harbin 150040, ChinaSchool of Forestry, Northeast Forestry University, Harbin 150040, ChinaObjects: 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.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022070337bacillus amyloliquefacienscellulasepurificationenzymatic property
spellingShingle Zhi ZHANG
Dongzhuo WEN
Lirong FENG
Shenglong ZHANG
Kexin YANG
Xiaotong ZHANG
Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4
Shipin gongye ke-ji
bacillus amyloliquefaciens
cellulase
purification
enzymatic property
title Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4
title_full Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4
title_fullStr Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4
title_full_unstemmed Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4
title_short Separation, Purification and Enzymatic Property of Cellulase Produced by Thermostable Bacillus amyloliquefaciens BA-DES4
title_sort separation purification and enzymatic property of cellulase produced by thermostable bacillus amyloliquefaciens ba des4
topic bacillus amyloliquefaciens
cellulase
purification
enzymatic property
url http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022070337
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