Molecular cloning and isolation of a recombinant alpha-Momorcharin in E. coli against Pyricularia oryzae
Từ khóa
DOI:
https://doi.org/10.32508/stdj.v26i1.4023Tóm tắt
Introduction: Alpha-Momorcharin (α-MMC) is a member of the ribosome-inactivating protein (RIP) family that has been widely used as an antitumor, antiviral and antifungal agent.
Methods: In this study, the codons of DNA encoding α-MMC were optimized for expression in E. coli and cloned into the pET-28a(+) vector. The protein was then expressed in E. coli strain BL21 (DE3) and purified by nickel affinity chromatography.
Results: Under IPTG induction, α-MMC was expressed at approximately 50% of the total protein, showing high-level recombinant protein expression in E. coli. A high amount of purified α-MMC (70 mg) was isolated from 1 L LB culture medium of E. coli BL21 (DE3) with approximately 95% purity. Interestingly, α-MMC inhibited the mycelial growth of Pyricularia oryzae in a concentration-dependent manner.
Conclusion: Using a microbial system for α-MMC expression provides a promising method for the design of a new agent against pathogens.
