Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation
In this study, a series of bacteria capable of degrading starch and cellulose were isolated from the aging flue-cured tobacco leaves. Remarkably, there was a thermophilic bacterium, Bacillus subtilis ZIM3, that can simultaneously degrade both starch and cellulose at a wide range of temperature and p...
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Frontiers Media S.A.
2020-07-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2020.01539/full |
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| author | Jingcheng Dai Aijun Dong Guoxi Xiong Yaqi Liu Yaqi Liu Md. Shahdat Hossain Md. Shahdat Hossain Md. Shahdat Hossain Shuangyuan Liu Shuangyuan Liu Na Gao Na Gao Shuyang Li Shuyang Li Jing Wang Jing Wang Dongru Qiu Dongru Qiu |
| author_facet | Jingcheng Dai Aijun Dong Guoxi Xiong Yaqi Liu Yaqi Liu Md. Shahdat Hossain Md. Shahdat Hossain Md. Shahdat Hossain Shuangyuan Liu Shuangyuan Liu Na Gao Na Gao Shuyang Li Shuyang Li Jing Wang Jing Wang Dongru Qiu Dongru Qiu |
| author_sort | Jingcheng Dai |
| collection | DOAJ |
| description | In this study, a series of bacteria capable of degrading starch and cellulose were isolated from the aging flue-cured tobacco leaves. Remarkably, there was a thermophilic bacterium, Bacillus subtilis ZIM3, that can simultaneously degrade both starch and cellulose at a wide range of temperature and pH values. Genome sequencing, comparative genomics analyses, and enzymatic activity assays showed that the ZIM3 strain expressed a variety of highly active plant biomass-degrading enzymes, such as the amylase AmyE1 and cellulase CelE1. The in vitro and PhoA-fusion assays indicated that these enzymes degrading complex plant biomass into fermentable sugars were secreted into ambient environment to function. Besides, the amylase and cellulase activities were further increased by three- to five-folds by using overexpression. Furthermore, a fermentation strategy was developed and the biodegradation efficiency of the starch and cellulose in the tobacco leaves were improved by 30–48%. These results reveal that B. subtilis ZIM3 and the recombinant strain exhibited high amylase and cellulase activities for efficient biodegradation of starch and cellulose in tobacco and could potentially be applied for industrial tobacco fermentation. |
| first_indexed | 2024-12-14T23:21:17Z |
| format | Article |
| id | doaj.art-59299ce194bb494a89514d708b5abb2d |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-14T23:21:17Z |
| publishDate | 2020-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-59299ce194bb494a89514d708b5abb2d2022-12-21T22:43:57ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2020-07-011110.3389/fmicb.2020.01539540567Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco FermentationJingcheng Dai0Aijun Dong1Guoxi Xiong2Yaqi Liu3Yaqi Liu4Md. Shahdat Hossain5Md. Shahdat Hossain6Md. Shahdat Hossain7Shuangyuan Liu8Shuangyuan Liu9Na Gao10Na Gao11Shuyang Li12Shuyang Li13Jing Wang14Jing Wang15Dongru Qiu16Dongru Qiu17Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaTechnology Research Center of China Tobacco Hubei Industry Co., Ltd., Wuhan, ChinaTechnology Research Center of China Tobacco Hubei Industry Co., Ltd., Wuhan, ChinaInstitute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaCollege of Fisheries and Life Science, Dalian Ocean University, Dalian, ChinaInstitute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaNational Institute of Biotechnology, Dhaka, BangladeshInstitute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaInstitute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaInstitute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaInstitute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaInstitute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaUniversity of Chinese Academy of Sciences, Beijing, ChinaIn this study, a series of bacteria capable of degrading starch and cellulose were isolated from the aging flue-cured tobacco leaves. Remarkably, there was a thermophilic bacterium, Bacillus subtilis ZIM3, that can simultaneously degrade both starch and cellulose at a wide range of temperature and pH values. Genome sequencing, comparative genomics analyses, and enzymatic activity assays showed that the ZIM3 strain expressed a variety of highly active plant biomass-degrading enzymes, such as the amylase AmyE1 and cellulase CelE1. The in vitro and PhoA-fusion assays indicated that these enzymes degrading complex plant biomass into fermentable sugars were secreted into ambient environment to function. Besides, the amylase and cellulase activities were further increased by three- to five-folds by using overexpression. Furthermore, a fermentation strategy was developed and the biodegradation efficiency of the starch and cellulose in the tobacco leaves were improved by 30–48%. These results reveal that B. subtilis ZIM3 and the recombinant strain exhibited high amylase and cellulase activities for efficient biodegradation of starch and cellulose in tobacco and could potentially be applied for industrial tobacco fermentation.https://www.frontiersin.org/article/10.3389/fmicb.2020.01539/fullamylasecellulaseheterogeneous expressionbacillustobacco |
| spellingShingle | Jingcheng Dai Aijun Dong Guoxi Xiong Yaqi Liu Yaqi Liu Md. Shahdat Hossain Md. Shahdat Hossain Md. Shahdat Hossain Shuangyuan Liu Shuangyuan Liu Na Gao Na Gao Shuyang Li Shuyang Li Jing Wang Jing Wang Dongru Qiu Dongru Qiu Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation Frontiers in Microbiology amylase cellulase heterogeneous expression bacillus tobacco |
| title | Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation |
| title_full | Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation |
| title_fullStr | Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation |
| title_full_unstemmed | Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation |
| title_short | Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation |
| title_sort | production of highly active extracellular amylase and cellulase from bacillus subtilis zim3 and a recombinant strain with a potential application in tobacco fermentation |
| topic | amylase cellulase heterogeneous expression bacillus tobacco |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2020.01539/full |
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