Precursor-directed production of erythromycin analogs by Saccharopolyspora erythraea

被引:25
作者
Frykman, S [1 ]
Leaf, T [1 ]
Carreras, C [1 ]
Licari, P [1 ]
机构
[1] Kosan Biosci Inc, Dept Proc Sci, Hayward, CA 94544 USA
关键词
chemobiosynthesis; diketide; erythraea; erythromycin; polyketide; thioester;
D O I
10.1002/bit.10086
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
Diketide N-acetylcysteamine (diketide NAC) thioester precursors were fed to 6-Deoxyerythronolide B synthase (DEBS) ketosynthase-1 inactivated (KS1 degrees) Saccharopolyspora erythraea strains to produce 13-substituted erythromycin analogs. This direct feeding process potentially represents a simplified production process over the current analog production system. Titers of these analogs were observed to increase linearly with the diketide concentration up to a precursor-specific saturation level. However, the rate of product formation was lower and the rate of diketide consumption higher with S. erythraea than was previously observed with a recombinant strain of Streptomyces coelicolor. Several strategies were pursued to address the issue of these high diketide consumption rates: (1) elucidation of the locale of diketide degradation, (2) addition of beta -oxidation inhibitors to the cultures, and (3) addition of a sacrificial diketide enantiomer to occupy putative degradative enzymes. Additionally, repeated addition of diketide to an S. erythraea KS1 degrees culture indicated that the titer of these erythromycin analogs is also currently limited by a shorter production period than observed during erythromycin synthesis by the parent strain. These results indicate potential avenues for expanding the use of this precursor-directed system from the generation of limited quantities of erythromycin analogs to a large-scale production system for these compounds. (C) 2001 John Wiley & Sons, Inc.
引用
收藏
页码:303 / 310
页数:8
相关论文
共 12 条
[1]   Production of 6-deoxy-13-cyclopropyl-erythromycin B by Saccharopolyspora erythraea NRRL 18643 [J].
Brown, MS ;
Dirlam, JP ;
McArthur, HAI ;
McCormick, EL ;
Morse, BK ;
Murphy, PA ;
O'Connell, TN ;
Pacey, M ;
Rescek, DM ;
Ruddock, J ;
Wax, RG .
JOURNAL OF ANTIBIOTICS, 1999, 52 (08) :742-747
[2]   Precursor-directed biosynthesis of erythromycin analogs by an engineered polyketide synthase [J].
Jacobsen, JR ;
Hutchinson, CR ;
Cane, DE ;
Khosla, C .
SCIENCE, 1997, 277 (5324) :367-369
[3]   Evidence for two catalytically independent clusters of active sites in a functional modular polyketide synthase [J].
Kao, CM ;
Pieper, R ;
Cane, DE ;
Khosla, C .
BIOCHEMISTRY, 1996, 35 (38) :12363-12368
[4]  
KIBWAGE IO, 1987, J ANTIBIOT, V40, P1
[5]   Precursor-directed biosynthesis of 6-deoxyerythronolide B analogs in Streptomyces coelicolor:: Understanding precursor effects [J].
Leaf, T ;
Cadapan, L ;
Carreras, C ;
Regentin, R ;
Ou, S ;
Woo, E ;
Ashley, G ;
Licari, P .
BIOTECHNOLOGY PROGRESS, 2000, 16 (04) :553-556
[6]   BIOSYNTHETIC INCORPORATION OF LABELED TETRAKETIDE INTERMEDIATES INTO DEHYDROCURVULARIN, A PHYTOTOXIN FROM ALTERNARIA-CINERARIAE, WITH ASSISTANCE OF BETA-OXIDATION INHIBITORS [J].
LI, Z ;
MARTIN, M ;
VEDERAS, JC .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1992, 114 (04) :1531-1533
[7]   Engineering broader specificity into an antibiotic-producing polyketide synthase [J].
Marsden, AFA ;
Wilkinson, B ;
Cortés, J ;
Dunster, NJ ;
Staunton, J ;
Leadlay, PF .
SCIENCE, 1998, 279 (5348) :199-202
[8]   REMARKABLY BROAD SUBSTRATE-SPECIFICITY OF A MODULAR POLYKETIDE SYNTHASE IN A CELL-FREE SYSTEM [J].
PIEPER, R ;
LUO, GL ;
CANE, DE ;
KHOSLA, C .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1995, 117 (45) :11373-11374
[9]   Metabolic routing towards polyhydroxyalkanoic acid synthesis in recombinant Escherichia coli (fadR):: Inhibition of fatty acid β-oxidation by acrylic acid [J].
Qi, QS ;
Steinbüchel, A ;
Rehm, BHA .
FEMS MICROBIOLOGY LETTERS, 1998, 167 (01) :89-94
[10]   IDENTIFICATION OF A SACCHAROPOLYSPORA-ERYTHRAEA GENE REQUIRED FOR THE FINAL HYDROXYLATION STEP IN ERYTHROMYCIN BIOSYNTHESIS [J].
STASSI, D ;
DONADIO, S ;
STAVER, MJ ;
KATZ, L .
JOURNAL OF BACTERIOLOGY, 1993, 175 (01) :182-189