Genome mining in Streptomyces coelicolor:: Molecular cloning and characterization of a new sesquiterpene synthase

被引：121

作者：

Lin, Xin ^{[1
]}

Hopson, Russell ^{[1
]}

Cane, David E. ^{[1
]}

机构：

[1] Brown Univ, Dept Chem, Providence, RI 02912 USA

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2006年 / 128卷 / 18期

关键词：

D O I：

10.1021/ja061292s

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The terpene synthase encoded by the SCO5222 (SC7E4.19) gene of Streptomyces coelicolor was cloned by PCR and expressed in Escherichia coli as an N-terminal-His6-tag protein. Incubation of the recombinant protein, SCO5222p, with farnesyl diphosphate (1, FPP) in the presence of Mg(II) gave a new sesquiterpene, (+)-epi-isozizaene (2), whose structure and stereochemistry were determined by a combination of 1H, 13C, COSY, HMQC, HMBC, and NOESY NMR. The steady-state kinetic parameters were kcat 0.049 ± 0.001 s-1 and a Km (FPP) of 147 ± 14 nM. Individual incubations of recombinant epi-isozizaene synthase with [1,1-2H2]FPP (1a), (1R)-[1-2H]-FPP (1b), and (1S)-[1-2H]-FPP (1c) and NMR analysis of the resulting deuterated epi-isozizaenes supported an isomerization-cyclization-rearrangement mechanism involving the intermediacy of (3R)-nerolidyl diphosphate (3). Copyright © 2006 American Chemical Society.

引用

页码：6022 / 6023

页数：2

共 14 条

[1] REARRANGEMENT OF ZIZAENE-RELATED SESQUITERPENES [J].

ANDERSEN, NH ;

SMITH, SE ;

OHTA, Y .

JOURNAL OF THE CHEMICAL SOCIETY-CHEMICAL COMMUNICATIONS, 1973, (13) :447-448

[2] Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2) [J].

Bentley, SD ;

Chater, KF ;

Cerdeño-Tárraga, AM ;

Challis, GL ;

Thomson, NR ;

James, KD ;

Harris, DE ;

Quail, MA ;

Kieser, H ;

Harper, D ;

Bateman, A ;

Brown, S ;

Chandra, G ;

Chen, CW ;

Collins, M ;

Cronin, A ;

Fraser, A ;

Goble, A ;

Hidalgo, J ;

Hornsby, T ;

Howarth, S ;

Huang, CH ;

Kieser, T ;

Larke, L ;

Murphy, L ;

Oliver, K ;

O'Neil, S ;

Rabbinowitsch, E ;

Rajandream, MA ;

Rutherford, K ;

Rutter, S ;

Seeger, K ;

Saunders, D ;

Sharp, S ;

Squares, R ;

Squares, S ;

Taylor, K ;

Warren, T ;

Wietzorrek, A ;

Woodward, J ;

Barrell, BG ;

Parkhill, J ;

Hopwood, DA .

NATURE, 2002, 417 (6885) :141-147

[3] Expression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesis [J].

Cane, DE ;

Watt, RM .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (04) :1547-1551

[4] PENTALENENE SYNTHASE - PURIFICATION, MOLECULAR-CLONING, SEQUENCING, AND HIGH-LEVEL EXPRESSION IN ESCHERICHIA-COLI OF A TERPENOID CYCLASE FROM STREPTOMYCES UC5319 [J].

CANE, DE ;

SOHNG, JK ;

LAMBERSON, CR ;

RUDNICKI, SM ;

WU, Z ;

LLOYD, MD ;

OLIVER, JS ;

HUBBARD, BR .

BIOCHEMISTRY, 1994, 33 (19) :5846-5857

[5] BIOSYNTHESIS OF PENTALENENE AND PENTALENOLACTONE [J].

CANE, DE ;

OLIVER, JS ;

HARRISON, PHM ;

ABELL, C ;

HUBBARD, BR ;

KANE, CT ;

LATTMAN, R .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1990, 112 (11) :4513-4524

[6]

GARSON MJ, 1979, CHEM SOC REV, P539

[7]

GURTLER H, 1994, J ANTIBIOT, V47, P434

[8] PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin [J].

Gust, B ;

Challis, GL ;

Fowler, K ;

Kieser, T ;

Chater, KF .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (04) :1541-1546

[9] Mechanism and stereochemistry of the germacradienol/germacrene D synthase of Streptomyces coelicolor A3(2) [J].

He, XF ;

Cane, DE .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (09) :2678-2679

[10] Primary metabolism and its control in streptomycetes: A most unusual group of bacteria [J].

Hodgson, DA .

ADVANCES IN MICROBIAL PHYSIOLOGY, VOL 42, 2000, 42 :47-238

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