Genomics-guided discovery of a new and significantly better source of anticancer natural drug FK228  

作　　者：Xiangyang Liu Feng Xie Leah B.Doughty Qi Wang Lixin Zhang Xueting Liu Yi-Qiang Cheng 

机构地区：[1]State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai,200237,PR China [2]UNT System College of Pharmacy,University of North Texas Health Science Center,Fort Worth,TX,76107,USA [3]CAS Key Laboratory of Pathogenic Microbiology and Immunology,Institute of Microbiology,Chinese Academy of Sciences,Beijing,100101,PR China [4]Department of Biological Sciences,University of Wisconsin-Milwaukee,Milwaukee,WI,53201,USA

出　　处：《Synthetic and Systems Biotechnology》2018年第4期268-274,共7页合成和系统生物技术（英文）

基　　金：the University of Wisconsin-Milwaukee Research Foundation and a Public Health Service grant(CA152212);the National Cancer Institute to YQC,the National Science Foundation of China(31430002,31770055,31570031);the Fundamental Research Funds for the Central Universities(22221818014);the Major Basic Program of the Natural Science Foundation of Shandong Province(ZR2017ZB0206)。

摘　　要：FK228 is an FDA-approved anticancer drug naturally produced by Chromobacterium violaceum No.968 up to 19 mg/L in a pilot industry-scale batch fermentation.Here we report a genomics-guided discovery of Burkholderia thailandensis MSMB43 as a new and significantly better source of FK228.The genome of B.thailandensis MSMB43 was found to contain a functional biosynthetic gene cluster highly homologous to that of FK228 in C.violaceum No.968,and the bacterium indeed produces authentic FK228.By simple fermentation in shaking flasks in a preferred M8 medium,B.thailandensis MSMB43 produced FK228 up to 67.7 mg/L;by fedbatch fermentation in a 20-L fermentor in M8 medium,B.thailandensis MSMB43 produced FK228 up to 115.9 mg/L,which is 95 fold higher than that of C.violaceum No.968 under the same laboratory fermentation conditions.RT-PCR analysis indicated that the high FK228 yield of B.thailandensis MSMB43 was due to high expression of biosynthetic genes,represented by Bth_depA,during the fermentation process.Further genetic manipulation resulted in a recombinant strain,B.thailandensis MSMB43/pBMTL3-tdpR,which harbors a broad host-range vector expressing the thailandepsin biosynthetic pathway regulatory gene tdpR.This engineered strain produced up to 168.5 mg/L of FK228 in fed-batch fermentation in a 20-L fermentor in M8 medium.Therefore,the wild-type B.thailandensis MSMB43 or its engineered derivative could potentially be a good starting point for an industrial process to improve FK228 production for its expanding use in therapy.

关 键 词：Burkholderia thailandensis MSMB43 Fermentation optimization FK228 Genome mining Natural product PRODUCTIVITY 

分 类 号：TQ9[化学工程]