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Effects of substrate feed rates on heterologous protein expression by Pichia pastoris in DO-stat fed-batch fermentation

Title
Effects of substrate feed rates on heterologous protein expression by Pichia pastoris in DO-stat fed-batch fermentation
Author
이은규
Keywords
Methylotrophic yeast; Induction with methanol; High-cell-density fermentation; DO-stat; Alpha-amylase
Issue Date
2003-09
Publisher
ELSEVIER SCIENCE INC
Citation
ENZYME AND MICROBIAL TECHNOLOGY, v. 33, issue. 4, page. 358-365
Abstract
Effects of regulated substrate feed on the cell growth and recombinant protein expression by the Pichia system were studied by applying either constant feed rates or variable feed rates in DO-stat fed-batch fermentation. Chimeric α-amylase (Amy1A/3D) that consists of 159 aa of one isozyme (Amy1A) of rice α-amylases followed by 251 aa of another (Amy3D) was used as a model protein. In the glycerol fed-batch (alcohol oxidase I derepression) phase, a step-decreasing glycerol feed control allowed a high growth rate (0.086 h−1) and high cell density (97.2 g DCW l−1). In the induction phase, methanol feed rate had a significant effect on the specific expression rate of Amy1A/3D. A constant feed rate of 1.0 ml l−1 min−1 was found to be optimum for a high specific expression rate of Amy1A/3D (0.091 mg g DCW−1 h−1). Under this condition, allowing a higher specific methanol consumption rate, a maximum concentration of 340 mg l−1 of Amy1A/3D and the expression yield of 2.16 mg Amy1A/3D g-methanol−1 were attained. This result strongly demonstrated that the efficiency of recombinant protein expression was directly dependent on methanol utilization in Pichia system.
URI
https://www.sciencedirect.com/science/article/pii/S0141022903001467https://repository.hanyang.ac.kr/handle/20.500.11754/156224
ISSN
0141-0229
DOI
10.1016/S0141-0229(03)00146-7
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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