摘要
【背景】从黄酒发酵液中分离的短乳杆菌2-34具有重吸收瓜氨酸的能力,可用于降低黄酒中的瓜氨酸从而减少氨基甲酸乙酯的形成,然而瓜氨酸重吸收机制的不明确阻碍了该菌的合理利用。【目的】通过确定短乳杆菌2-34中的瓜氨酸转运蛋白编码基因,为其在黄酒中的利用提供理论依据。【方法】以pRSFDuet-1和pETDuet-1为表达载体,通过多顺反子串联表达系统及双质粒表达系统,在大肠杆菌C43(DE3)中表达短乳杆菌2-34精氨酸脱亚胺途径中瓜氨酸代谢相关蛋白:精氨酸降解酶ArcD和ADI、瓜氨酸降解酶OTC及膜蛋白DcuC和AO antiporter。【结果】重组表达大肠杆菌发酵过程中可利用精氨酸形成瓜氨酸,但表达了膜蛋白DcuC和AO antiporter的重组菌发酵液中瓜氨酸含量较低。【结论】短乳杆菌2-34中DcuC和AOantiporter均具有吸收瓜氨酸的功能,且DcuC活性更高。
[Background]Citrulline and ethyl carbamate are hazards generated during rice wine fermentation.From rice wine fermentation broth Lactobacillus brevis 2-34 is isolated with the potential to reabsorb citrulline.[Objective]To identify genes encoding citrulline transporters in this strain will help its effective application in rice wine fermentation.[Methods]With vectors pRSFDuet-1 and pETDuet-1,citrulline metabolism-related enzymes of L.brevis 2-34,including arginine/ornithine antiporter,arginine deaminase,ornithine carbamyltransferase,as well as the two putative citrulline transporters,C4-dicarboxylate anaerobic carrier and AO antiporter,were expressed in Escherichia coli C43(DE3).[Results]Recombinant E.coli had similar citrulline reabsorption ability as L.brevis 2-34.Both putative transporters could absorb extracellular citrulline,and the transporting capability of DcuC was much stronger.[Conclusion]DcuC and AO antiporter are citrulline transporters in L.brevis 2-34.
作者
刘晓慧
李晓敏
禹伟
吴殿辉
陆健
LIU Xiao-Hui;LI Xiao-Min;YU Wei;WU Dian-Hui;LU Jian(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi,Jiangsu 214122,China;National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi,Jiangsu 214122,ChinaSchool of Biotechnology,Jiangnan University,Wuxi,Jiangsu 214122,China;Jiangsu Provincial Research Center for Bioactive Product Processing Technology,Wuxi,Jiangsu 214122,China)
出处
《微生物学通报》
CAS
CSCD
北大核心
2019年第9期2249-2257,共9页
Microbiology China
基金
国家自然科学基金(3170101535)~~
关键词
黄酒
短乳杆菌
瓜氨酸重吸收
重组表达
Rice wine
Lactobacillus brevis
Citrulline reabsorption
Recombinant expression
