Codon optimization of Saccharomyces cerevisiae mating factor alpha prepro-leader to improve recombinant protein production in Pichia pastoris = 피키아 파스토리스에의 재조합 단백질 생산 향상을 위한 사카로마이세스 유래의 분비서열 코돈 최적화

Abstract

Objectives: To evaluate different codon optimization parameters on the Saccharomyces cerevisiae-derived mating factor α prepro-leader sequence (MFLS) to improve Candida antarctica lipase B (CAL-B) secretory production in Pichia pastoris. Results: Codon optimization based on the individual codon usage (ICU) and codon context (CC) design parameters enhanced secretory production of CAL-B to 7U/ml and 12U/ml, respectively. Only 3U/ml was obtained with the wild type sequence while the sequence optimized using both ICU and CC objectives showed intermediate performance of 10U/ml. These results clearly show that CC is the most relevant parameter for the codon optimization of MFLS in P. pastoris, and there is no synergistic effect achieved by considering both ICU and CC together. Conclusion: The CC optimized MFLS increased secretory protein production of CAL-B in P. pastoris by fourfold.