Title: | Codon optimization of Saccharomyces cerevisiae mating factor alpha prepro-leader to improve recombinant protein production in Pichia pastoris |
Author(s): | Ahn J; Jang MJ; Ang KS; Lee H; Choi ES; Lee DY; |
Address: | "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. Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, Singapore. 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" |
DOI: | 10.1007/s10529-016-2203-3 |
ISSN/ISBN: | 1573-6776 (Electronic) 0141-5492 (Linking) |
Abstract: | "OBJECTIVES: 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. 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. CONCLUSION: The CC optimized MFLS increased secretory protein production of CAL-B in P. pastoris by fourfold" |
Keywords: | Codon/*genetics Mating Factor/genetics Pichia/*genetics/*metabolism Recombinant Proteins/*genetics/*metabolism Saccharomyces cerevisiae/*genetics Synthetic Biology Codon optimization Codon-pair context Individual codon usage Mating factor alpha prepro-lea; |
Notes: | "MedlineAhn, Jungoh Jang, Min-Jung Ang, Kok Siong Lee, Hongweon Choi, Eui-Sung Lee, Dong-Yup eng Netherlands 2016/10/25 Biotechnol Lett. 2016 Dec; 38(12):2137-2143. doi: 10.1007/s10529-016-2203-3. Epub 2016 Sep 9" |