Expanding tryptophan-containing cyclodipeptide synthase spectrum by identification of nine members from Streptomyces strains
Expanding tryptophan-containing cyclodipeptide synthase spectrum by identification of nine members from Streptomyces strains
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DOI：
10.1007/s00253-018-8908-6
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发表时间：
2018-05-01
影响因子：
5
通讯作者：
Li, Shu-Ming
中科院分区：
文献类型：
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作者：
Liu, Jing;Yu, Huili;Li, Shu-Ming
环二肽合成酶(CDPS)通常由200-300个氨基酸残基组成，主要存在于细菌中。他们劫持了核糖体机器中的氨基-tRNA，以形成环二肽。本研究将8株链霉菌的9个新的CDPS基因克隆到pET28a载体中，并在大肠杆菌中进行了表达。对分离出的产物进行结构鉴定，鉴定出一个环-L-Trp-L-Leu，两个环-L-Trp-L-Pro和三个环-L-Trp-L-Trp合成酶。其他3个CDPS以环-L-色氨酸-L-丙氨酸或环-L-色氨酸-L-酪氨酸为主要环二肽。总产物产量为46~211 mg/L。我们的发现代表了来自放线杆菌的CDPs家族的罕见例子，这些放线杆菌形成了各种含有色氨酸的环二肽。此外，本研究突出了含色氨酸环二肽生物合成的微生物机制的潜力，并为这些CDPS基因与其他修饰基因在合成生物学中的进一步结合提供了有效的实验基础。
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.