Codon optimization of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> mating factor alpha prepro-leader to improve recombinant protein production in <Emphasis Type="Italic">Pichia pastoris</Emphasis> |
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| Authors: | Jungoh Ahn Min-Jung Jang Kok Siong Ang Hongweon Lee Eui-Sung Choi Dong-Yup Lee |
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| Institution: | 1.Biotechnology Process Engineering Center,Korea Research Institute of Bioscience and Biotechnology (KRIBB),Cheongju,Korea;2.Department of Biotechnology Process Engineering,University of Science and Technology (UST),Daejeon,South Korea;3.Department of Chemical and Biomolecular Engineering,National University of Singapore,Singapore,Singapore;4.Bioprocessing Technology Institute,Agency for Science, Technology and Research (A*STAR),Singapore,Singapore;5.Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute,National University of Singapore,Singapore,Singapore |
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| Abstract: | ObjectivesTo 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.ResultsCodon 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.ConclusionThe CC optimized MFLS increased secretory protein production of CAL-B in P. pastoris by fourfold. |
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