A Pichia pastoris fermentation strategy for enhancing the heterologous expression of an Escherichia coli phytase

被引:78
作者
Chen, CC
Wu, PH
Huang, CT
Cheng, KJ
机构
[1] Natl Taiwan Univ, Inst Microbiol & Biochem, Sect 4, Taipei 106, Taiwan
[2] Natl Taiwan Univ, Dept Biochem Sci, Sect 4, Taipei 106, Taiwan
[3] Acad Sinica, Inst Bioagr Sci, Taipei 115, Taiwan
关键词
appA; methanol induction; Pichia pastoris; phytase;
D O I
10.1016/j.enzmictec.2004.05.007
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The Escherichia coli phytase gene appA was highly expressed in the methylotrophic yeast Pichia pastoris under the control of the AOX1 promoter. Replacement of culture medium with fresh medium in order to remove repressing glycerol and metabolic wastes prior to methanol induction significantly improved phytase expression. The phytase activity level was enhanced from 118 to 204U/ml at the flask scale and 1880-4946 U/ml for high cell-density fermentation, respectively, by appropriately modifying the medium composition and fermentation strategy. Most of the protein in the culture supernatant was recombinant phytase, the enzyme characteristics of which were similar to native E. coli phytase. (C) 2004 Elsevier Inc. All rights reserved.
引用
收藏
页码:315 / 320
页数:6
相关论文
共 33 条
[1]   A novel staining method for detecting phytase activity [J].
Bae, HD ;
Yanke, LJ ;
Cheng, KJ ;
Selinger, LB .
JOURNAL OF MICROBIOLOGICAL METHODS, 1999, 39 (01) :17-22
[2]  
Cereghino JL, 2000, FEMS MICROBIOL REV, V24, P45, DOI 10.1016/S0168-6445(99)00029-7
[3]   PHYTIC ACID INTERACTIONS IN FOOD SYSTEMS [J].
CHERYAN, M .
CRC CRITICAL REVIEWS IN FOOD SCIENCE AND NUTRITION, 1980, 13 (04) :297-335
[4]   THE COMPLETE NUCLEOTIDE-SEQUENCE OF THE ESCHERICHIA-COLI GENE APPA REVEALS SIGNIFICANT HOMOLOGY BETWEEN PH 2.5 ACID-PHOSPHATASE AND GLUCOSE-1-PHOSPHATASE [J].
DASSA, J ;
MARCK, C ;
BOQUET, PL .
JOURNAL OF BACTERIOLOGY, 1990, 172 (09) :5497-5500
[5]   Heterologous protein production in methylotrophic yeasts [J].
Gellissen, G .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2000, 54 (06) :741-750
[6]   PURIFICATION AND CHARACTERIZATION OF PHYTASE FROM COTYLEDONS OF GERMINATING SOYBEAN SEEDS [J].
GIBSON, DM ;
ULLAH, AHJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1988, 260 (02) :503-513
[7]   Characterization and overproduction of the Escherichia coli appA encoded bifunctional enzyme that exhibits both phytase and acid phosphatase activities [J].
Golovan, S ;
Wang, GR ;
Zhang, J ;
Forsberg, CW .
CANADIAN JOURNAL OF MICROBIOLOGY, 2000, 46 (01) :59-71
[8]   High-level expression and stabilization of recombinant human chitinase produced in a continuous constitutive Pichia pastoris expression system [J].
Goodrick, JC ;
Xu, M ;
Finnegan, R ;
Schilling, BM ;
Schiavi, S ;
Hoppe, H ;
Wan, NC .
BIOTECHNOLOGY AND BIOENGINEERING, 2001, 74 (06) :492-497
[9]   PURIFICATION AND CHARACTERIZATION OF 2 PHYTASES FROM ESCHERICHIA-COLI [J].
GREINER, R ;
KONIETZNY, U ;
JANY, KD .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1993, 303 (01) :107-113
[10]   Production of recombinant proteins by methylotrophic yeasts [J].
Hollenberg, CP ;
Gellissen, G .
CURRENT OPINION IN BIOTECHNOLOGY, 1997, 8 (05) :554-560