Expanding tryptophan-containing cyclodipeptide synthase spectrum by identification of nine members from Streptomyces strains
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Cyclodipeptide synthases (CDPSs) comprise normally 200–300 amino acid residues and are mainly found in bacteria. They hijack aminoacyl-tRNAs from the ribosomal machinery for cyclodipeptide formation. In this study, nine new CDPS genes from eight Streptomyces strains were cloned into pET28a vector and expressed in Escherichia coli. Structural elucidation of the isolated products led to the identification of one cyclo-l-Trp-l-Leu, two cyclo-l-Trp-l-Pro, and three cyclo-l-Trp-l-Trp synthases. Other three CDPSs produce cyclo-l-Trp-l-Ala or cyclo-l-Trp-l-Tyr as the major cyclodipeptide. Total product yields of 46 to 211 mg/L E. coli culture were obtained. Our findings represent rare examples of CDPS family derived from actinobacteria that form various tryptophan-containing cyclodipeptides. Furthermore, this study highlights the potential of the microbial machinery for tryptophan-containing cyclodipeptide biosynthesis and provides valid experimental basis for further combination of these CDPS genes with other modification genes in synthetic biology.
KeywordsAminoacyl t-RNA Diketopiperazine Cyclodipeptide synthase Streptomyces Tryptophan-containing cyclodipeptide
We thank ARS Culture Collection (NRRL) for providing Streptomyces strains, S. Newel, and R. Kraut (University Marburg) for taking NMR and MS spectra.
The Bruker micrOTOF QIII mass spectrometer was financially supported in part by a grant from the Deutsche Forschungsgemeinschaft (INST 160/620-1 to S.-M. L.). J.L. and H.Y are scholarship recipients of China Scholarship Council (201608310118 and 201306220024).
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Conflict of interest
The authors declare that they have no conflict of interest.
Human and animal rights
This article does not contain any studies with human participants or animals performed by any of the authors.
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