Thioesterase domain swapping of a linear polyketide tautomycetin with a macrocyclic polyketide pikromycin in Streptomyces sp. CK4412.
Tautomycetin (TMC) is a linear polyketide metabolite produced by Streptomyces sp. CK4412 that has been reported to possess multiple biological functions including T cell-specific immunosuppressive and anticancer activities that occur through a mechanism of differential inhibition of protein phosphatases such as PP1, PP2A, and SHP2. We previously reported the characterization of the entire TMC biosynthetic gene cluster constituted by multifunctional type I polyketide synthase (PKS) assembly and suggested that the linear form of TMC could be generated via free acid chain termination by a narrow TMC thioesterase (TE) pocket. The modular nature of the assembly presents a unique opportunity to alter or interchange the native biosynthetic domains to produce targeted variants of TMC. Herein, we report swapping of the TMC TE domain sequence with the exact counterpart of the macrocyclic polyketide pikromycin (PIK) TE. PIK TE-swapped Streptomyces sp. CK4412 mutant produced not only TMC, but also a cyclized form of TMC, implying that the bioengineering based in vivo custom construct can be exploited to produce engineered macrolactones with new structural functionality.
Life Sciences Institute and Departments of Medicinal Chemistry, Chemistry, and Microbiology and Immunology, University of Michigan, Ann Arbor, MI, 48109-2216, USA.
Ashootosh
A
Tripathi
Department of Biological Engineering, Inha University, Incheon, 402-751, Korea.
Si-Sun
SS
Choi
Life Sciences Institute and Departments of Medicinal Chemistry, Chemistry, and Microbiology and Immunology, University of Michigan, Ann Arbor, MI, 48109-2216, USA. davidhs@umich.edu.
David H
DH
Sherman
Department of Biological Engineering, Inha University, Incheon, 402-751, Korea. eungsoo@inha.ac.kr.
Eung-Soo
ES
Kim
D053898:Biosynthetic Pathways / Q000235:genetics; D060846:Cell Engineering; D005663:Furans / Q000737:chemistry* / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D018942:Macrolides / Q000737:chemistry*; D048630:Polyketide Synthases / Q000235:genetics; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*; D013869:Thiolester Hydrolases / Q000737:chemistry / Q000378:metabolism
D005663:Furans; D008055:Lipids; D018942:Macrolides; C058744:tautomycetin; D048630:Polyketide Synthases; D013869:Thiolester Hydrolases; C003145:picromycin
Domain swapping; Linear polyketide; Pikromycin; Tautomycetin; Thioesterase