Choeng Yong-Hoon, Yang Ji-Yeon, Delcroix Gaetan, Kim Yoon-Jung, Chang Yong-Keun, Hong Soon-Kwang
Division of Bioscience and Bioinformatics, Myongji University, Yongin, Korea.
J Microbiol Biotechnol. 2007 Oct;17(10):1675-81.
The pseudodisaccharide salbostatin, which consists of valienamine linked to 2-amino-1,5-anhydro-2-deoxyglucitol, is a strong trehalase inhibitor. From our Streptomyces albus ATCC 21838 genomic library, we identified thirty-nine ORFs in a 40-kb gene cluster. Twenty-one genes are supposed to be a complete set of modules responsible for the salbostatin biosynthesis. Through sequence analysis of the gene cluster, some of the upstream gene products (SalB, SalC, SalD, SalE, and SalF) revealed functional resemblance with trehalose biosynthetic enzymes. On the basis of this rationale, we isolated the five genes (salB, salC, salD, salE, and salF) from the S. albus ATCC 21838 and cloned them into the expression vector pWHM3. We demonstrated the noticeable expression and accumulation of trehalose, using only the five upstream biosynthetic gene cluster of salbostatin, in the transformed Streptomyces lividans TK24. Finally, 490 mg/l trehalose was produced by fermentation of the transformant with sucrosedepleted R2YE media.
由缬氨胺与2-氨基-1,5-脱水-2-脱氧葡萄糖醇相连组成的假二糖salbostatin是一种强效海藻糖酶抑制剂。从我们的白色链霉菌ATCC 21838基因组文库中，我们在一个40 kb的基因簇中鉴定出39个开放阅读框（ORF）。21个基因被认为是负责salbostatin生物合成的一套完整模块。通过对该基因簇的序列分析，一些上游基因产物（SalB、SalC、SalD、SalE和SalF）显示出与海藻糖生物合成酶的功能相似性。基于这一原理，我们从白色链霉菌ATCC 21838中分离出五个基因（salB、salC、salD、salE和salF），并将它们克隆到表达载体pWHM3中。我们仅使用salbostatin的五个上游生物合成基因簇，就在转化的淡紫链霉菌TK24中证明了海藻糖的显著表达和积累。最后，用不含蔗糖的R2YE培养基发酵该转化体，产生了490 mg/l的海藻糖。
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