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Codon optimization of Saccharomyces cerevisiae mating factor alpha prepro-leader to improve recombinant protein production in Pichia pastoris

Ahn, J.; Jang, M.-J.; Ang, K.S.; Lee, H.; Choi, E.-S.; Lee, D.-Y.

Biotechnology Letters 38(12): 2137-2143

2016


ISSN/ISBN: 1573-6776
PMID: 27613154
DOI: 10.1007/s10529-016-2203-3
Accession: 057434705

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. Codon optimization based on the individual codon usage (ICU) and codon context (CC) design parameters enhanced secretory production of CAL-B to 7 U/ml and 12 U/ml, respectively. Only 3 U/ml was obtained with the wild type sequence while the sequence optimized using both ICU and CC objectives showed intermediate performance of 10 U/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. The CC optimized MFLS increased secretory protein production of CAL-B in P. pastoris by fourfold.

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