Jungoh Ahn1,2, Min-Jung Jang1, Kok Siong Ang3, Hongweon Lee1,2, Eui-Sung Choi1,2, Dong-Yup Lee4,5,6. 1. Biotechnology Process Engineering Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 30 Yeongudanji-ro, Ochang-eup, Cheongwon-gu, Cheongju, 363-883, Korea. 2. Department of Biotechnology Process Engineering, University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon, South Korea. 3. Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, Singapore. 4. Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, Singapore. cheld@nus.edu.sg. 5. Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01, Centros, Singapore, 138668, Singapore. cheld@nus.edu.sg. 6. 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.
Abstract
OBJECTIVES: 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. CONCLUSION: The CC optimized MFLS increased secretory protein production of CAL-B in P. pastoris by fourfold.
OBJECTIVES: 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. CONCLUSION: The CC optimized MFLS increased secretory protein production of CAL-B in P. pastoris by fourfold.
Authors: Juan J Barrero; Jason C Casler; Francisco Valero; Pau Ferrer; Benjamin S Glick Journal: Microb Cell Fact Date: 2018-10-12 Impact factor: 5.328
Authors: Juan J Barrero; Alejandro Pagazartaundua; Benjamin S Glick; Francisco Valero; Pau Ferrer Journal: N Biotechnol Date: 2020-10-09 Impact factor: 5.079