Rational design of a bimodular model system for the investigation of heterocyclization in nonribosomal peptide biosynthesis

被引：49

作者：

Duerfahrt, T

Eppelmann, K

Müller, R

Marahiel, MA

机构：

[1] Univ Marburg, Fachbereich Chem Biochem, D-35043 Marburg, Germany

[2] Gesell Biotechnol Forsch mbH, D-38124 Braunschweig, Germany

来源：

CHEMISTRY & BIOLOGY | 2004年 / 11卷 / 02期

关键词：

D O I：

10.1016/j.chembiol.2004.01.013

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Cyclization (Cy) domains in NRPS catalyze the heterocyclization of cysteine and serine/threonine to thiazoline and oxazoline rings. A model system consisting of the first two modules of bacitracin synthetase A fused to the thioesterase (Te) domain of tyrocidine synthetase was constructed (BaCA1-2-Te) and shown to be active in production of the heterocyclic IleCys(thiazoline). Based on this model system, the feasibility of Cy domain module fusions was investigated by replacing the BacA2 Cy-A-PCP-module with modules of MbtB and MtaD from the biosynthesis systems of mycobactin and myxothiazol, revealing the formation of novel heterocyclic dipeptides. To dissect the reaction sequence of the Cy domain in peptide bond formation and heterocyclization, several residues of the BaCA1-2-Te Cy domain were analyzed by mutagenesis. Two mutants exhibited formation of the noncyclic dipeptide, providing clear evidence for the independence of condensation and cyclization.

引用

页码：261 / 271

页数：11

共 38 条

[1] Aminoacyl-CoAs as probes of condensation domain selectivity in nonribosomal peptide synthesis
Belshaw, PJ
Walsh, CT
Stachelhaus, T
[J]. SCIENCE, 1999, 284 (5413) : 486 - 489
[2] Mutational analysis of the C-domain in nonribosomal peptide synthesis
Bergendahl, V
Linne, U
Marahiel, MA
[J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 2002, 269 (02): : 620 - 629
[3] Structural basis for the activation of phenylalanine in the non-ribosomal biosynthesis of gramicidin S
Conti, E
Stachelhaus, T
Marahiel, MA
Brick, P
[J]. EMBO JOURNAL, 1997, 16 (14) : 4174 - 4183
[4] Dipeptide formation on engineered hybrid peptide synthetases
Doekel, S
Marahiel, MA
[J]. CHEMISTRY & BIOLOGY, 2000, 7 (06): : 373 - 384
[5] BlmIII and BImIV nonribosornal peptide synthetase-catalyzed biosynthesis of the bleomycin bithiazole moiety involving both in Cis and in trans aminoacylation
Du, LC
Chen, M
Zhang, Y
Shen, B
[J]. BIOCHEMISTRY, 2003, 42 (32) : 9731 - 9740
[6] Du LC, 2000, FEMS MICROBIOL LETT, V189, P171, DOI 10.1111/j.1574-6968.2000.tb09225.x
[7] Construction of hybrid peptide synthetases for the production of α-L-aspartyl-L-phenylalanine, a precursor for the high-intensity sweetener aspartame
Duerfahrt, T
Doekel, S
Sonke, T
Quaedflieg, PJLM
Marahiel, MA
[J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 2003, 270 (22): : 4555 - 4563
[8] Epothilons A and B: Antifungal and cytotoxic compounds from Sorangium cellulosum (Myxobacteria) - Production, physico-chemical and biological properties
Gerth, K
Bedorf, N
Hofle, G
Irschik, H
Reichenbach, H
[J]. JOURNAL OF ANTIBIOTICS, 1996, 49 (06) : 560 - 563
[9] MYXOTHIAZOL, AN ANTIBIOTIC FROM MYXOCOCCUS-FULVUS (MYXOBACTERALES) .1. CULTIVATION, ISOLATION, PHYSICOCHEMICAL AND BIOLOGICAL PROPERTIES
GERTH, K
IRSCHIK, H
REICHENBACH, H
TROWITZSCH, W
[J]. JOURNAL OF ANTIBIOTICS, 1980, 33 (12) : 1474 - 1479
[10] STRUCTURAL CHARACTERIZATION OF BACITRACIN COMPONENTS BY FRIT-FAST ATOM BOMBARDMENT (FAB) LIQUID-CHROMATOGRAPHY MASS-SPECTROMETRY (LC/MS)
IKAI, Y
OKA, H
HAYAKAWA, J
HARADA, K
SUZUKI, M
[J]. JOURNAL OF ANTIBIOTICS, 1992, 45 (08) : 1325 - 1334

← 1 2 3 4 →