文献类型：期刊文献

英文题名：Toward Improvement of Erythromycin A Production in an Industrial Saccharopolyspora erythraea Strain via Facilitation of Genetic Manipulation with an Artificial attB Site for Specific Recombination

作者：Wu, Jiequn[1,2];Zhang, Qinglin[1,2];Deng, Wei[1];Qian, Jiangchao[2];Zhang, Siliang[2];Liu, Wen[1]

机构：[1]Chinese Acad Sci, Shanghai Inst Organ Chem, State Key Lab Bioorgan & Nat Prod Chem, Shanghai 200032, Peoples R China;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2011

卷号：77

期号：21

起止页码：7508

外文期刊名：APPLIED AND ENVIRONMENTAL MICROBIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000296568200009)】；

基金：This work was supported in part by grants from the National Natural Science Foundation (20832009 and 20921091), the National Basic Research Program (973 program, 2012CB721100 and 2010CB833200), the Chinese Academy of Sciences (KJCX2-YW-201), and the Science and Technology Commission of Shanghai Municipality (09QH1402700) of China.

语种：英文

摘要：Large-scale production of erythromycin A (Er-A) relies on the organism Saccharopolyspora erythraea, in which lack of a typical attB site largely impedes the application of phage Phi C31 integrase-mediated recombination into site-specific engineering. We herein report construction of an artificial attB site in an industrial S. erythraea strain, HL3168 E3, in an effort to break the bottleneck previously encountered during genetic manipulation mainly from homologous or unpredictable nonspecific integration. Replacement of a cryptic gene, nrps1-1, with a cassette containing eight attB DNA sequences did not affect the high Er-producing ability, setting the stage for precisely engineering the industrial Er-producing strain for foreign DNA introduction with a reliable conjugation frequency. Transfer of either exogenous or endogenous genes of importance to Er-A biosynthesis, including the S-adenosylmethionine synthetase gene for positive regulation, vhb for increasing the oxygen supply, and two tailoring genes, eryK and eryG, for optimizing the biotransformation at the late stage, was achieved by taking advantage of this facility, allowing systematic improvement of Er-A production as well as elimination of the by-products Er-B and Er-C in fermentation. The strategy developed here can generally be applicable to other strains that lack the attB site.