Genomics-guided discovery of a new and significantly better source of anticancer natural drug FK228
| Autor: | Feng Xie, Xiangyang Liu, Lixin Zhang, Xueting Liu, Yi-Qiang Cheng, Qi Wang, Leah B. Doughty |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
FK228 lcsh:Biotechnology 030106 microbiology Biomedical Engineering Burkholderia thailandensis MSMB43 Applied Microbiology and Biotechnology Article Natural product law.invention 03 medical and health sciences chemistry.chemical_compound Structural Biology law Genome mining lcsh:TP248.13-248.65 Gene cluster Genetics lcsh:QH301-705.5 Productivity Regulator gene biology Burkholderia thailandensis biology.organism_classification Fermentation optimization lcsh:Biology (General) chemistry Biochemistry Recombinant DNA Fermentation Chromobacterium violaceum Bacteria |
| Zdroj: | Synthetic and Systems Biotechnology, Vol 3, Iss 4, Pp 268-274 (2018) Synthetic and Systems Biotechnology |
| ISSN: | 2405-805X |
| DOI: | 10.1016/j.synbio.2018.10.011 |
| Popis: | 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 fed-batch 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. Keywords: Burkholderia thailandensis MSMB43, Fermentation optimization, FK228, Genome mining, Natural product, Productivity |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|