Journal
BIOTECHNOLOGY LETTERS
Volume 38, Issue 12, Pages 2137-2143Publisher
SPRINGER
DOI: 10.1007/s10529-016-2203-3
Keywords
Codon optimization; Codon-pair context; Individual codon usage; Mating factor alpha prepro-leader sequence; Pichia pastoris; Saccharomyces cerevisiae; Synthetic biology
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Funding
- National University of Singapore
- Biomedical Research Council of A*STAR (Agency for Science, Technology and Research), Singapore
- Grants from the Global R&D project program [N011500017]
- Ministry of Trade, Industry and Energy (MOTIE), Republic of Korea
- Next-Generation BioGreen 21 Program (SSAC) [PJ01109405]
- Rural Development Administration, Republic of Korea
- KRIBB Research Initiative Program
- Ministry of Trade, Industry & Energy (MOTIE), Republic of Korea [N0000677] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- National Research Council of Science & Technology (NST), Republic of Korea [KGM4631633] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- Rural Development Administration (RDA), Republic of Korea [PJ011094052016] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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To evaluate different codon optimization parameters on the Saccharomyces cerevisiae-derived mating factor alpha 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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