Metabolic engineering of Bacillus amyloliquefaciens for efficient production ofα-glucosidase inhibitor1-deoxynojirimycin  

作　　者：Xujie Li Meng Zhang Yu Lu Ningyang Wu Jian’gang Chen Zhixia Ji Yangyang Zhan Xin Ma Junyong Chen Dongbo Cai Shouwen Chen 

机构地区：[1]State Key Laboratory of Biocatalysis and Enzyme Engineering,Environmental Microbial Technology Center of Hubei Province,College of Life Sciences,Hubei University,Wuhan,430062,China [2]State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan,430070,China [3]Department of Urology,Zhuhai People’s Hospital(Zhuhai Hospital Affiliated to Jinan University),Zhuhai,519000,China [4]Wuhan Jun’an Biotechnology Co.,Ltd.,Wuhan,430070,China

出　　处：《Synthetic and Systems Biotechnology》2023年第3期378-385,共8页合成和系统生物技术（英文）

基　　金：Key research and development program of Hubei Province(2022BBA0031);the Key Science and Technology Innovation Project of Hubei Province(2021BAD001);Wuhan Science and Technology Project(2020020602012124);Knowledge Innovation Program of Wuhan-Shuguang Project(2022020801020334).

摘　　要：Owing to the feature of strongα-glucosidase inhibitory activity,1-deoxynojirimycin(1-DNJ)has broad application prospects in areas of functional food,biomedicine,etc.,and this research wants to construct an efficient strain for 1-DNJ production,basing on Bacillus amyloliquefaciens HZ-12.Firstly,using the temperature-sensitive shuttle plasmid T2(2)-Ori,gene ptsG in phosphotransferase system(PTS)was weakened by homologous recombination,and non-PTS pathway was strengthened by deleting its repressor gene iolR,and 1-DNJ yield of resultant strain HZ-S2 was increased by 4.27-fold,reached 110.72 mg/L.Then,to increase precursor fructose-6-phosphate(F-6-P)supply,phosphofructokinase was weaken,fructose phosphatase GlpX and 6-phosphate glucose isomerase Pgi were strengthened by promoter replacement,moreover,regulator gene nanR was deleted,1-DNJ yield was further increased to 267.37 mg/L by 2.41-fold.Subsequently,promoter of 1-DNJ synthetase cluster was optimized,as well as 5′-UTRs of downstream genes in synthetase cluster,and 1-DNJ produced by the final strain reached 478.62 mg/L.Last but not the least,1-DNJ yield of 1632.50 mg/L was attained in 3 L fermenter,which was the highest yield of 1-DNJ reported to date.Taken together,our results demonstrated that metabolic engineering was an effective strategy for 1-DNJ synthesis,this research laid a foundation for industrialization of functional food and drugs based on 1-DNJ.

关 键 词：1-DEOXYNOJIRIMYCIN Bacillus amyloliquefaciens Metabolic engineering Glucose transport pathway Fructose 6-phosphate 1-DNJ synthetase Gene cluster 

分 类 号：Q93[生物学—微生物学]