Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazard

The combustion of excessive starch in tobacco leaves leads to more harmful substances, adversely affecting the sensory properties of tobacco and posing significant risks to human health. Therefore, there is an urgent need to develop specific amylases targeting tobacco starch to address these issues....

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Main Authors: Zongchen Han, Jie Hao, Dian Zou, Zhikang Sun, Zekun Zhang, Chenqi Niu, Qi Lu, Kuo Huang, Changwen Ye, Xuetuan Wei
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1603337/full
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author Zongchen Han
Jie Hao
Dian Zou
Zhikang Sun
Zekun Zhang
Chenqi Niu
Qi Lu
Kuo Huang
Changwen Ye
Xuetuan Wei
author_facet Zongchen Han
Jie Hao
Dian Zou
Zhikang Sun
Zekun Zhang
Chenqi Niu
Qi Lu
Kuo Huang
Changwen Ye
Xuetuan Wei
author_sort Zongchen Han
collection DOAJ
description The combustion of excessive starch in tobacco leaves leads to more harmful substances, adversely affecting the sensory properties of tobacco and posing significant risks to human health. Therefore, there is an urgent need to develop specific amylases targeting tobacco starch to address these issues. In this study, 5 different α-amylase genes were selected for recombinant expression in Bacillus amyloliquefaciens BAX-5, and the amyA(LC) (derived from Bacillus amyloliquefaciens MK10163) was confirmed to be the optimal gene. Then, the α-amylase activity was further increased by screening host bacteria BAX-5 and signal peptides SP003 (derived from the dacB gene of Bacillus subtilis 168). Subsequently, the α-amylase properties were characterized, such as temperature tolerance, pH tolerance and metal ion. Through replacement of culture medium, the recombinant strain BAX-5/PT-17SP003amyA(LC) produced the maximum α-amylase activity of 904.91 IU/mL, which was about 4 times higher than that of the original culture medium. Finally, the α-amylase Amy (LC) was applied to the enzyme treatment of tobacco leaves, and the evaluation results showed that α-amylase Amy (LC) could play a positive role in reducing damage and enhancing quality of cigarettes. This research provides a novel enzymatic resource for the development of amylases, and it has enormous market potential and application value.
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institution Kabale University
issn 1664-302X
language English
publishDate 2025-07-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Microbiology
spelling doaj-art-037e7de557fa4bcc891b3ce05b215f3d2025-08-20T03:50:06ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.16033371603337Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazardZongchen Han0Jie Hao1Dian Zou2Zhikang Sun3Zekun Zhang4Chenqi Niu5Qi Lu6Kuo Huang7Changwen Ye8Xuetuan Wei9Zhengzhou Tobacco Research Institute of China National Tobacco Corporation, Zhengzhou, ChinaInner Mongolia Kunming Cigarettes LLC, Hohhot, ChinaState Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, ChinaInner Mongolia Kunming Cigarettes LLC, Hohhot, ChinaInner Mongolia Kunming Cigarettes LLC, Hohhot, ChinaZhengzhou Tobacco Research Institute of China National Tobacco Corporation, Zhengzhou, ChinaInstitute of Agro-Products Processing and Nuclear Agricultural Technology, Hubei Academy of Agricultural Sciences, Wuhan, ChinaZhengzhou Tobacco Research Institute of China National Tobacco Corporation, Zhengzhou, ChinaZhengzhou Tobacco Research Institute of China National Tobacco Corporation, Zhengzhou, ChinaState Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, ChinaThe combustion of excessive starch in tobacco leaves leads to more harmful substances, adversely affecting the sensory properties of tobacco and posing significant risks to human health. Therefore, there is an urgent need to develop specific amylases targeting tobacco starch to address these issues. In this study, 5 different α-amylase genes were selected for recombinant expression in Bacillus amyloliquefaciens BAX-5, and the amyA(LC) (derived from Bacillus amyloliquefaciens MK10163) was confirmed to be the optimal gene. Then, the α-amylase activity was further increased by screening host bacteria BAX-5 and signal peptides SP003 (derived from the dacB gene of Bacillus subtilis 168). Subsequently, the α-amylase properties were characterized, such as temperature tolerance, pH tolerance and metal ion. Through replacement of culture medium, the recombinant strain BAX-5/PT-17SP003amyA(LC) produced the maximum α-amylase activity of 904.91 IU/mL, which was about 4 times higher than that of the original culture medium. Finally, the α-amylase Amy (LC) was applied to the enzyme treatment of tobacco leaves, and the evaluation results showed that α-amylase Amy (LC) could play a positive role in reducing damage and enhancing quality of cigarettes. This research provides a novel enzymatic resource for the development of amylases, and it has enormous market potential and application value.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1603337/fulltobacco starchα-amylaseBacillus amyloliquefaciensexpression optimizationapplication evaluation
spellingShingle Zongchen Han
Jie Hao
Dian Zou
Zhikang Sun
Zekun Zhang
Chenqi Niu
Qi Lu
Kuo Huang
Changwen Ye
Xuetuan Wei
Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazard
Frontiers in Microbiology
tobacco starch
α-amylase
Bacillus amyloliquefaciens
expression optimization
application evaluation
title Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazard
title_full Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazard
title_fullStr Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazard
title_full_unstemmed Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazard
title_short Efficient expression of a novel α-amylase for reduction of tobacco starch and smoke hazard
title_sort efficient expression of a novel α amylase for reduction of tobacco starch and smoke hazard
topic tobacco starch
α-amylase
Bacillus amyloliquefaciens
expression optimization
application evaluation
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1603337/full
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