CcpA mutants infuence selective carbon source utilization by changing interactions with target genes in Bacillus licheniformis  

作　　者：Yupeng Zhang Youran Li Fengxu Xiao Hanrong Wang Liang Zhang Zhongyang Ding Sha Xu Zhenghua Gu Guiyang Shi 

机构地区：[1]Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,People’s Republic of China [2]National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,1800 Lihu Avenue,Wuxi,Jiangsu Province 214122,People’s Republic of China [3]Jiangsu Provincial Research Center for Bioactive Product Processing Technology,Jiangnan University,Wuxi,People’s Republic of China

出　　处：《Systems Microbiology and Biomanufacturing》2022年第1期193-207,共15页系统微生物学与生物制造（英文）

基　　金：This work was supported by National Key Research&Development Program of China(2018YFA0900504,2020YFA0907700 and 2018YFA0900300);the National Natural Foundation of China(31401674);the National First-Class Discipline Program of Light Industry Technology and Engineering(LITE2018-22);the Topnotch Academic Programs Project of Jiangsu Higher Education Institutions.

摘　　要：The gram-positive bacterium Bacillus licheniformis exhibits obvious selective utilization on carbon sources.This process is mainly governed by the global regulator catabolite control protein A(CcpA),which can recognize and bind to multiple target genes that are widely distributed in metabolic pathways.Although the DNA-binding domain of CcpA has been predicted,the infuence of key amino acids on target gene recognition and binding has yet to be uncovered.In this study,the impact of Lys31,Ile42 and Leu56 on in vitro protein–DNA interactions and in vivo carbon source selective utilization was investigated.The results showed that alanine substitution of Lys31 and Ile42,located within the 3rd helices of the DNAbinding domain,signifcantly weakened the binding strength between CcpA and target genes.These mutations also lead to alleviated repression of xylose utilization in the presence of glucose.On the other hand,the Leu56Arg mutant in the 4th helices exhibited enhanced binding afnity compared with that of the wild-type one.When this mutant was used to replace the native one in B.licheniformis cells,the selective utilization of glucose over xylose increased.This study provides a new strategy for understanding the relationship between the function and structure of regulatory proteins.This study also used a new strategy was used to regulate carbon source utilization beyond CCR engineering.

关 键 词：CCPA Bacillus licheniformis MUTAGENESIS Xylose utilization 

分 类 号：O62[理学—有机化学]