Fed-batch high-cell-density fermentation strategies for Pichia pastoris growth and production

Abstract

Pichia pastoris is extensively used to produce various heterologous proteins. Amounts of biopharmaceutical drugs and industrial enzymes have been successfully produced by fed-batch high-cell-density fermentation (HCDF) of this cell factory. High levels of cell mass in defined media can be easily achieved and therefore large quantities of recombinant proteins with enhanced activities and lower costs can be obtained through HCDF technology. A robust HCDF process makes a successful transition to commercial production. Recently, efforts have been made to increase the heterologous protein production and activity by the HCDF of P. pastoris. However, challenges around selecting a suitable HCDF strategy exist. The high-level expression of a specific protein in P. pastoris is still, at least in part, limited by optimizing the methanol feeding strategy. Here, we review the progress in developments and applications of P. pastoris HCDF strategies for enhanced expression of recombinant proteins. We focus on the methanol induction strategies for efficient fed-batch HCDF in bioreactors, mainly focusing on various stat-induction strategies, co-feeding, and the limited induction strategy. These processes control strategies have opened the door for expressing foreign proteins in P. pastoris and are expected to enhance the production of recombinant proteins.

Keywords:

• Pichia pastoris
• high-cell-density fermentation strategy
• stat-induction
• co-substrate feeding
• limited induction
• process control and monitoring

Acknowledgments

The work was partially carried out in the Biotechnology Core Laboratory, NIH/NIDDK. We would like to thank Dr. Joseph Shiloach for his kind assistance also we like to thank the NIH fellows editorial board (FEB) for editorial assistance.

Disclosure statement

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China under Grant Numbers 81573325 and 31270796; CAMS Initiative for Innovative Medicine under Grant Numbers CAMS-2017-I2M-2–004 and CAMS-2016-I2M-2–002; National Infrastructure of Microbial Resources under Grant Number NIMR-2018–3; Fundamental Research Funds for the Central Universities under Grant Numbers 2016ZX350054 and 2017PT35001; National Mega-project for Innovative Drugs under Grant Numbers 2018ZX09711001-006–001 and 2012ZX09301002-001–005; Beijing Natural Science Foundation under Grant Number 7174320.