Epothilone C macrolactonization and hydrolysis are catalyzed by the isolated thioesterase domain of epothilone polyketide synthase

被引：53
作者：
Boddy, CN
Schneider, TL
Hotta, K
Walsh, CT
Khosla, C [1 ]
机构：
[1] Stanford Univ, Dept Chem Engn, Stanford, CA 94305 USA
[2] Stanford Univ, Dept Chem, Stanford, CA 94305 USA
[3] Stanford Univ, Dept Biochem, Stanford, CA 94305 USA
[4] Harvard Univ, Sch Med, Dept Biol Chem & Mol Pharmacol, Boston, MA 02115 USA
来源：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2003年 / 125卷 / 12期
关键词：
D O I：
10.1021/ja0298646
中图分类号：
O6 [化学];
学科分类号：
0703 ;
摘要：
Epothilone C is produced by the combined action of one nonribosomal peptide synthetase (NRPS) and nine polyketide synthase (PKS) modules in a multienzyme system. The final step in the biosynthesis is the thioesterase (TE)-catalyzed cyclorelease of epothilone from the EpoF protein. It has been unclear whether isolated PKS TE domains could exhibit macrolactonization activity. Here we demonstrate that the excised epothilone TE domain can catalyze the efficient cyclization of the N-acetylcysteamine thioester of seco-epothilone C to generate epothilone C (kcat/KM = 0.41 ± 0.03 min-1 mM-1). The TE domain also catalyzes the hydrolysis of both the N-acetylcysteamine thioester of seco-epothilone C (kcat = 0.087 ± 0.005 min-1, KM = 291 ± 53 μM) and that of the epothilone C (kcat = 0.67 ± 0.01 min-1, KM = 117 ± 5 μM) to form seco-epothilone C. Copyright © 2003 American Chemical Society.
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页码：3428 / 3429
页数：2
相关论文
共 32 条
[1]   Epothilones and related structures - a new class of microtubule inhibitors with potent in vivo antitumor activity [J].
Altmann, KH ;
Wartmann, M ;
O'Reilly, T .
BIOCHIMICA ET BIOPHYSICA ACTA-REVIEWS ON CANCER, 2000, 1470 (03) :M79-M91
[2]  
BOLLAG DM, 1995, CANCER RES, V55, P2325
[3]   Epothilone biosynthesis: assembly of the methylthiazolylcarboxy starter unit on the EpoB subunit [J].
Chen, HW ;
O'Connor, S ;
Cane, DE ;
Walsh, CT .
CHEMISTRY & BIOLOGY, 2001, 8 (09) :899-912
[4]   REPOSITIONING OF A DOMAIN IN A MODULAR POLYKETIDE SYNTHASE TO PROMOTE SPECIFIC CHAIN CLEAVAGE [J].
CORTES, J ;
WIESMANN, KEH ;
ROBERTS, GA ;
BROWN, MJB ;
STAUNTON, J ;
LEADLAY, PF .
SCIENCE, 1995, 268 (5216) :1487-1489
[5]   Epothilons A and B: Antifungal and cytotoxic compounds from Sorangium cellulosum (Myxobacteria) - Production, physico-chemical and biological properties [J].
Gerth, K ;
Bedorf, N ;
Hofle, G ;
Irschik, H ;
Reichenbach, H .
JOURNAL OF ANTIBIOTICS, 1996, 49 (06) :560-563
[6]   Mechanism and specificity of the terminal thioesterase domain from the erythromycin polyketide synthase [J].
Gokhale, RS ;
Hunziker, D ;
Cane, DE ;
Khosla, C .
CHEMISTRY & BIOLOGY, 1999, 6 (02) :117-125
[7]   Epothilone A and B - Novel 16-membered macrolides with cytotoxic activity: Isolation, crystal structure, and conformation in solution [J].
Hofle, GH ;
Bedorf, N ;
Steinmetz, H ;
Schomburg, D ;
Gerth, K ;
Reichenbach, H .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION IN ENGLISH, 1996, 35 (13-14) :1567-1569
[8]   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
[9]   MANIPULATION OF MACROLIDE RING SIZE BY DIRECTED MUTAGENESIS OF A MODULAR POLYKETIDE SYNTHASE [J].
KAO, CM ;
LUO, GL ;
KATZ, L ;
CANE, DE ;
KHOSLA, C .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1995, 117 (35) :9105-9106
[10]   Engineered biosynthesis of structurally diverse tetraketides by a trimodular polyketide synthase [J].
Kao, CM ;
Luo, GL ;
Katz, L ;
Cane, DE ;
Khosla, C .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1996, 118 (38) :9184-9185
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