Scaling-up Fermentation of Pichia pastoris to demonstration-scale using new methanol-feeding strategy and increased air pressure instead of pure oxygen supplement

被引：82

作者：

Liu, Wan-Cang ^{[1
,2
]}

Gong, Ting ^{[1
,2
]}

Wang, Qing-Hua ^{[2
,3
]}

Liang, Xiao ^{[2
,3
]}

Chen, Jing-Jing ^{[1
,2
]}

Zhu, Ping ^{[1
,2
,3
]}

机构：

[1] Chinese Acad Med Sci, Inst Mat Med, State Key Lab Bioact Subst & Funct Nat Med, 1 Xian Nong Tan St, Beijing 100050, Peoples R China

[2] Peking Union Med Coll, 1 Xian Nong Tan St, Beijing 100050, Peoples R China

[3] Chinese Acad Med Sci, Key Lab Biosynth Nat Prod, Natl Hlth & Family Planning Commiss, Inst Mat Med, 1 Xian Nong Tan St, Beijing 100050, Peoples R China

来源：

SCIENTIFIC REPORTS | 2016年 / 6卷

关键词：

RECOMBINANT PROTEIN; YARROWIA-LIPOLYTICA; SACCHAROMYCES-CEREVISIAE; BETA-GALACTOSIDASE; LIPASE PRODUCTION; CULTURES; BATCH; EXPRESSION; YIELD; PURIFICATION;

D O I：

10.1038/srep18439

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Scaling-up of high-cell-density fermentation (HCDF) of Pichia pastoris from the lab or pilot scale to the demonstration scale possesses great significance because the latter is the final technological hurdle in the decision to go commercial. However, related investigations have rarely been reported. In this paper, we study the scaling-up processes of a recombinant P. pastoris from the pilot (10 to 100-L) to the demonstration (1,000-L) scales, which can be used to convert 7-beta-xylosyl-10-deacetyltaxol into 10-deacetyltaxol by the beta-xylosidase for semi-synthesis of Taxol. We demonstrated that a pure oxygen supplement can be omitted from the HCDF if the super atmospheric pressure was increased from 0.05 to 0.10 +/- 0.05 MPa, and we developed a new methanol feeding biomass-stat strategy (0.035 mL/g/h) with 1% dissolved oxygen and 100 g/L initial induction biomass (dry cell weight). The scaling-up was reproducible, and the best results were obtained from the 1,000-L scale, featuring a shorter induction time and the highest enzyme activities and productions, respectively. The specific growth and specific production rates were also determined. This study lays a solid foundation for the commercial preparation of 10-deacetyltaxol through the recombinant yeast. It also provides a successful paradigm for scaling-up HCDF of P. pastoris to the demonstration scale.

引用

页数：12