Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis

This study was conducted to increase the enzymatic activity of <i>Bacillus licheniformis</i> XS-4, which was isolated from the traditional fermented mash of Xianshi soy sauce. The mutation was induced by atmospheric and room-temperature plasma (ARTP), and a mutant strain, mut80, was obta...

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Main Authors: Andong Zhang, Yudong Ma, Yue Deng, Zhiwei Zhou, Yue Cao, Bin Yang, Jing Bai, Qun Sun
Format: Article
Language:English
Published: MDPI AG 2023-06-01
Series:Foods
Subjects:
Online Access:https://www.mdpi.com/2304-8158/12/12/2381
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author Andong Zhang
Yudong Ma
Yue Deng
Zhiwei Zhou
Yue Cao
Bin Yang
Jing Bai
Qun Sun
author_facet Andong Zhang
Yudong Ma
Yue Deng
Zhiwei Zhou
Yue Cao
Bin Yang
Jing Bai
Qun Sun
author_sort Andong Zhang
collection DOAJ
description This study was conducted to increase the enzymatic activity of <i>Bacillus licheniformis</i> XS-4, which was isolated from the traditional fermented mash of Xianshi soy sauce. The mutation was induced by atmospheric and room-temperature plasma (ARTP), and a mutant strain, mut80, was obtained. mut80 exhibited significant increases in protease and amylase activity by 90.54% and 143.10%, respectively, and the enhanced enzymatic activities were stably maintained after 20 consecutive incubations. Re-sequencing analysis of mut80 revealed that the mutation sites were located in 1518447(AT-T) and 4253106(G-A) in its genome, which was involved in the metabolic pathways of amino acids. The expression of the protease synthetic gene (<i>aprX</i>) increased 1.54 times, while that of the amylase gene (<i>amyA</i>) increased 11.26 times, as confirmed via RT-qPCR. Using ARTP mutagenesis, the present study proposes a highly efficient microbial resource with enhanced protease and amylase activity provided by <i>B. licheniformis</i>, which can potentially be used to improve the efficiency of traditional soy sauce fermentation.
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spelling doaj.art-4187a6c3cb6848db81080d216a3c01172023-11-18T10:24:54ZengMDPI AGFoods2304-81582023-06-011212238110.3390/foods12122381Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP MutagenesisAndong Zhang0Yudong Ma1Yue Deng2Zhiwei Zhou3Yue Cao4Bin Yang5Jing Bai6Qun Sun7Key Laboratory of Bio-Resources and Eco-Environment of the Ministry of the Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaKey Laboratory of Bio-Resources and Eco-Environment of the Ministry of the Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaSchool of China Alcoholic Drinks, Luzhou Vocational and Technical College, Luzhou 646000, ChinaKey Laboratory of Bio-Resources and Eco-Environment of the Ministry of the Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaKey Laboratory of Bio-Resources and Eco-Environment of the Ministry of the Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaKey Laboratory of Bio-Resources and Eco-Environment of the Ministry of the Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaKey Laboratory of Bio-Resources and Eco-Environment of the Ministry of the Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaKey Laboratory of Bio-Resources and Eco-Environment of the Ministry of the Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaThis study was conducted to increase the enzymatic activity of <i>Bacillus licheniformis</i> XS-4, which was isolated from the traditional fermented mash of Xianshi soy sauce. The mutation was induced by atmospheric and room-temperature plasma (ARTP), and a mutant strain, mut80, was obtained. mut80 exhibited significant increases in protease and amylase activity by 90.54% and 143.10%, respectively, and the enhanced enzymatic activities were stably maintained after 20 consecutive incubations. Re-sequencing analysis of mut80 revealed that the mutation sites were located in 1518447(AT-T) and 4253106(G-A) in its genome, which was involved in the metabolic pathways of amino acids. The expression of the protease synthetic gene (<i>aprX</i>) increased 1.54 times, while that of the amylase gene (<i>amyA</i>) increased 11.26 times, as confirmed via RT-qPCR. Using ARTP mutagenesis, the present study proposes a highly efficient microbial resource with enhanced protease and amylase activity provided by <i>B. licheniformis</i>, which can potentially be used to improve the efficiency of traditional soy sauce fermentation.https://www.mdpi.com/2304-8158/12/12/2381<i>Bacillus licheniformis</i> mut80ARTPprotease and amylasetraditional soy saucefermentation
spellingShingle Andong Zhang
Yudong Ma
Yue Deng
Zhiwei Zhou
Yue Cao
Bin Yang
Jing Bai
Qun Sun
Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis
Foods
<i>Bacillus licheniformis</i> mut80
ARTP
protease and amylase
traditional soy sauce
fermentation
title Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis
title_full Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis
title_fullStr Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis
title_full_unstemmed Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis
title_short Enhancing Protease and Amylase Activities in <i>Bacillus licheniformis</i> XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis
title_sort enhancing protease and amylase activities in i bacillus licheniformis i xs 4 for traditional soy sauce fermentation using artp mutagenesis
topic <i>Bacillus licheniformis</i> mut80
ARTP
protease and amylase
traditional soy sauce
fermentation
url https://www.mdpi.com/2304-8158/12/12/2381
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