Journal of Northern Agriculture ›› 2025, Vol. 53 ›› Issue (2): 22-35.doi: 10.12190/j.issn.2096-1197.2025.02.03

• Molecular biology·Physiology and biochemistry • Previous Articles     Next Articles

Screening,identification,and enzyme production conditions optimization of a highly efficient starch-degrading bacterial

YU Shanjie1, ZHANG Aojie1, ZHENG Jie1, YU Yao1, BAI Xuehan1, WANG Pingping2, GUO Chunsheng3, ZHANG Lixin1   

  1. 1. College of Life Sciences,Northwest A&F University,Yangling 712100,China;
    2. Shaanxi Tobacco Science Research Institute,Xi′an 710000,China;
    3. Inner Mongolia Kunming Cigarette Co.,Ltd.,Hohhot 010020,China
  • Received:2024-10-29 Online:2025-04-20 Published:2025-09-08

Abstract: 【Objective】To screen a bacterial strain capable of efficiently degrading starch and to optimize its enzyme production conditions,providing novel methods and insights for starch degradation in tobacco leaves.【Methods】Starch-degrading bacterial strains were isolated from high-quality flue-cured tobacco varieties(Yunyan 97,Yunyan 99,and Qinyan 99) leaves in Shaanxi Province. A dominant strain was identified through preliminary screening based on the Hc value,secondary screening for amylase activity,morphological observation,physiological and biochemical tests,and molecular biological identification. Single-factor experiments and orthogonal experiments were conducted to optimize the enzyme production conditions of the dominant strain.【Results】A total of 250 starch-degrading bacterial strains were isolated,and Bacillus amyloliquefaciens ZJ-189 was identified as the dominant tarch-degrading strain with an amylase activity of 53.07 U/mL.Following ultraviolet(UV) mutagenesis,124 mutant strains were obtained from B. amyloliquefaciens ZJ-189. Through secondary screening for amylase activity,the highly efficient starch-degrading strain ZJ189-UV11 was selected,exhibiting an amylase activity of 74.42 U/mL,a 40.23% increase compared to the starter strain,with stable genetic characteristics. Orthogonal experiments revealed that the optimal culture conditions for strain ZJ189-UV11 after 24 h were a temperature of 42 ℃,pH of 7.5,agitation speed of 200 r/min,and an inoculation ratio of 1.2%.【Conclusion】Bacillus amyloliquefaciens ZJ-189 is identifid as a dominant strain for starch degradation. The culture conditions affecting the growth and amylase activity of the highly efficient mutant strain ZJ189-UV11 are optimized through single-factor experiments and orthogonal experiments.

Key words: Tobacco leaves, Bacillus amyloliquefaciens, UV mutagenesis, Enzyme production conditions optimization

CLC Number: 

  • TS414