Phenotypic and genomic insights into mutant with high nattokinase-producing activity induced by carbon ion beam irradiation of Bacillus subtilis

Yanan Sheng; Shuang Zhang; Xintong Li; Shicheng Wang; Tao Liu; Changyuan Wang; Lei Yan

doi:10.1016/j.ijbiomac.2024.132398

Phenotypic and genomic insights into mutant with high nattokinase-producing activity induced by carbon ion beam irradiation of Bacillus subtilis

Int J Biol Macromol. 2024 May 15;271(Pt 1):132398. doi: 10.1016/j.ijbiomac.2024.132398. Online ahead of print.

Authors

Yanan Sheng¹, Shuang Zhang², Xintong Li², Shicheng Wang², Tao Liu², Changyuan Wang¹, Lei Yan³

Affiliations

¹ College of Food, Heilongjiang Bayi Agricultural University, Daqing 163319, PR China.
² Heilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang 163319, China.
³ College of Food, Heilongjiang Bayi Agricultural University, Daqing 163319, PR China; Heilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang 163319, China. Electronic address: hekouyanlei@gmail.com.

PMID: 38754670
DOI: 10.1016/j.ijbiomac.2024.132398

Abstract

Nattokinase (NK) is found in fermented foods and has high fibrinolytic activity, which makes it promising for biological applications. In this study, a mutant strain (Bacillus subtilis ZT-S1, 5529.56 ± 183.59 U/mL) with high NK-producing activity was obtained using ¹²C⁶⁺ heavy ion beam mutagenesis for the first time. The surface morphology of B. subtilis is also altered by changes in functional groups caused by heavy ion beams. Furthermore, B. subtilis ZT-S1 required more carbon and nitrogen sources and reached stabilization phase later. Comparative genome analysis revealed that most of the mutant implicated genes (oppA, appA, kinA, spoIIP) were related to spore formation. And the affected rpoA is related to the synthesis of the NK-coding gene aprE. In addition, the B. subtilis ZT-S1 obtained by mutagenesis had good genetic stability. This study further explores the factors affecting NK activity and provides a promising microbial resource for NK production in commercial applications.

Keywords: (12)C(6+) heavy ions; Comparative genome; Mutagenized.