Enhancement of recombinant BmK AngM1 production in Pichia pastoris by regulating gene dosage, co-expressing with chaperones and fermenting in fed-batch mode. Issue 6 (3rd June 2017)

• Record Type: Journal Article
• Authors: Wang, Qing-Hua; Liang, Lan; Liu, Wan-Cang; Gong, Ting; Chen, Jing-Jing; Hou, Qi; Yang, Jin-Ling; Zhu, Ping
• Is Part Of: Journal of Asian natural products research. Volume 19:Issue 6(2017)
• Page Start: 581
• Page End: 594
• Publication Date: 2017-06-03
• Subjects: BmK AngM1 -- Pichia pastoris -- gene dosage -- molecular chaperones -- high-density fermentation; Materia medica -- Asia -- Periodicals; Pharmacognosy -- Asia -- Periodicals; Medicinal plants -- Asia -- Periodicals; Medicine, Oriental -- Asia -- Periodicals; 615.321095
• DOI: 10.1080/10286020.2017.1311872
• Languages: English
• ISSNs: 1028-6020

Abstract

Abstract: The scorpion peptide BmK AngM1 was reported to exhibit evident analgesic effect, but its yield by extraction from scorpion venom limits the research and application. The heterologous expression of BmK AngM1 was achieved in Pichia pastoris in our previous study. In order to realize high-level expression of recombinant BmK AngM1 (rBmK AngM1), the gene dosage of BmK AngM1 was optimized in engineered strains. The yield of rBmK AngM1 in the four-copy strain reached up to 100 mg/L, which was further enhanced to 190 mg/L by co-expressing with chaperones of PDI, BiP, and HAC1. Moreover, the yield of rBmK AngM1 was up to 1200 mg/L by high-density fermentation in 10 L fermenter. Finally, 360 mg rBmK AngM1 was purified from 1 L cultures by a two-step purification method. The efficient and convenient techniques presented in this study could facilitate further scale-up for industrial production of rBmK AngM1. Abstract :