Heterologous expression and antitumor activity analysis of syringolin from Pseudomonas syringae pv. syringae B728a

Microbial Cell Factories
Fan HuangLiqiu Xia

Abstract

Syringolin, synthesized by a mixed non-ribosomal peptide synthetase/polyketide synthetase in Pseudomonas syringae pv. syringae (Pss) B728a, is a novel eukaryotic proteasome inhibitor. Meanwhile, directly modifying large fragments in the PKS/NRPS gene cluster through traditional DNA engineering techniques is very difficult. In this study, we directly cloned the syl gene cluster from Pss B301D-R via Red/ET recombineering to effectively express syringolin in heterologous hosts. A 22 kb genomic fragment containing the sylA-sylE gene cluster was cloned into the pASK vector, and the obtained recombinant plasmid was transferred into Streptomyces coelicolor and Streptomyces lividans for the heterologous expression of syringolin. Transcriptional levels of recombinant syl gene in S. coelicolor M145 and S. lividans TK24 were evaluated via RT-PCR and the production of syringolin compounds was detected via LC-MS analysis. The extracts of the engineered bacteria showed cytotoxic activity to B16, 4T1, Meth-A, and HeLa tumor cells. It is noteworthy that the syringolin displayed anticancer activity against C57BL/6 mice with B16 murine melanoma tumor cells. Together, our results herein demonstrate the potential of syrinolin as effective antitumo...Continue Reading

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Citations

Jan 9, 2019·Natural Product Reports·Liujie HuoRolf Müller
Aug 28, 2019·Natural Product Reports·Maksym Myronovskyi, Andriy Luzhetskyy
Jun 15, 2019·Natural Product Reports·Jia Jia ZhangBradley S Moore
Jan 15, 2021·Current Opinion in Biotechnology·Hahk-Soo Kang, Eung-Soo Kim
Jan 25, 2020·Trends in Biotechnology·Muhammad Nazeer AbbasiAiying Li
Jul 31, 2021·Frontiers in Bioengineering and Biotechnology·Wenfang WangYinhua Lu

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Datasets Mentioned

BETA
AJ548826

Methods Mentioned

BETA
PCR
electrophoresis
ELISA
lavage

Software Mentioned

Bruker Compass

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