标题:3-Hydroxylation of the polycyclic tetramate macrolactam in the biosynthesis of antifungal HSAF from Lysobacter enzymogenes C3~(??)
作者:Yaoyao Li;Justin Huffman;Yu Li;Liangcheng Du;Yuemao Shen
作者机构:[Li, Y] School of Pharmaceutical Sciences, Shandong University, Jinan 250100, China;[ Huffman, J] Department of Chemistry, University of Nebraska-Linc 更多
通讯作者:Du, LC
通讯作者地址:[Du, LC]Univ Nebraska, Dept Chem, Lincoln, NE 68588 USA.
来源:MedChemComm
出版年:2012
卷:3
期:8
页码:982-986
DOI:10.1039/c2md20026k
摘要:HSAF isolated from Lysobacter enzymogenes is an antifungal natural product with a new mode of action. The polycyclic tetramate macrolactam (PTM) carries a 3-hydroxyl group on the ornithine residue, which is a feature common to nearly all PTM type natural products found in phylogenetically diverse organisms. A previous gene disruption experiment indicated that the sterol desaturase (SD) gene, which is clustered with the central polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS), was involved in the 3-hydroxylation. However, the mechanism for this hydroxylation had not been established. Here, we determined the structure of the main metabolite accumulated in the SD mutant, which is 3-dehydroxy HSAF (3-deOH-HSAF). This compound lost the antifungal activity against Penicillium avellaneum, showing the crucial role of the 3-hydroxyl group. We then expressed the SD gene in E. coli. Upon feeding 3-deOH-HSAF, E. coli produced HSAF. When the SD enzyme extract was incubated with 3-deOH-HSAF in the presence of NADPH, HSAF was also produced. The results demonstrated that the SD gene encodes a 3-hydroxylase of the HSAF carbon chain. In addition, the data support that the 3-hydroxylation step is likely a post-PKS-NRPS event in the HSAF biosynthetic pathway. Finally, we co-expressed the ferredoxin reductase (FNR) gene, which is also clustered with the PKS-NRPS gene, with the SD gene in E. coli. The results showed that FNR significantly enhanced the conversion of 3-deOH-HSAF into HSAF. Together, the study established the mechanism for the installation of this common functionality that is important to the activity of the PTM-type natural products.
收录类别:SCOPUS;SCIE
WOS核心被引频次:22
Scopus被引频次:23
资源类型:期刊论文
原文链接:https://www.scopus.com/inward/record.uri?eid=2-s2.0-84864367139&doi=10.1039%2fc2md20026k&partnerID=40&md5=5d1438421077f5a8504cec07be5db7f2
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