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


CONCLUSIONS
The CC optimized MFLS increased secretory protein production of CAL-B in P. pastoris by fourfold.
Biotechnology Process Engineering Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 30 Yeongudanji-ro, Ochang-eup, Cheongwon-gu, Cheongju, 363-883, Korea.|Department of Biotechnology Process Engineering, University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon, South Korea.
Jungoh
J
Ahn
Biotechnology Process Engineering Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 30 Yeongudanji-ro, Ochang-eup, Cheongwon-gu, Cheongju, 363-883, Korea.
Min-Jung
MJ
Jang
Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, Singapore.
Kok Siong
KS
Ang
Biotechnology Process Engineering Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 30 Yeongudanji-ro, Ochang-eup, Cheongwon-gu, Cheongju, 363-883, Korea.|Department of Biotechnology Process Engineering, University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon, South Korea.
Hongweon
H
Lee
Biotechnology Process Engineering Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 30 Yeongudanji-ro, Ochang-eup, Cheongwon-gu, Cheongju, 363-883, Korea.|Department of Biotechnology Process Engineering, University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon, South Korea.
Eui-Sung
ES
Choi
Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, Singapore. cheld@nus.edu.sg.|Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01, Centros, Singapore, 138668, Singapore. cheld@nus.edu.sg.|Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, 28 Medical Drive, Singapore, 117456, Singapore. cheld@nus.edu.sg.
Dong-Yup
DY
Lee
D003062:Codon / Q000235:genetics*; D000072235:Mating Factor / Q000235:genetics; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D058615:Synthetic Biology
D003062:Codon; D011994:Recombinant Proteins; D000072235:Mating Factor
Codon optimization; Codon-pair context; Individual codon usage; Mating factor α prepro-leader sequence; Pichia pastoris; Saccharomyces cerevisiae; Synthetic biology