Reaction mechanism and stereochemistry of
gamma-hexachlorocyclohexane dehydrochlorinase LinA

J 2001

Reaction mechanism and stereochemistry of gamma-hexachlorocyclohexane dehydrochlorinase LinA

TRANTÍREK, Lukáš; Kamila HYNKOVÁ; Yuji NAGATA; Alexey MURZIN; Alena ANSORGOVÁ et. al.

Basic information

Original name

Reaction mechanism and stereochemistry of gamma-hexachlorocyclohexane dehydrochlorinase LinA

Authors

TRANTÍREK, Lukáš; Kamila HYNKOVÁ; Yuji NAGATA; Alexey MURZIN; Alena ANSORGOVÁ; Vladimír SKLENÁŘ and Jiří DAMBORSKÝ

Edition

Journal of Biological Chemistry, 2001

Other information

Language

English

Type of outcome

Article in a journal

Field of Study

10600 1.6 Biological sciences

Country of publisher

United States of America

Confidentiality degree

is not subject to a state or trade secret

References:

URL

RIV identification code

RIV/00216224:14310/01:00004145

Organization unit

Faculty of Science

UT WoS

000167474900011

Changed: 28/5/2003 22:04, prof. Mgr. Jiří Damborský, Dr.

Abstract

In the original language

g-hexachlorocyclohexane dehydrochlorinase (LinA) catalyses the initial steps in the biotransformation of the important insecticide g-hexachlorocyclohexane (g-HCH) by the soil bacterium Sphingomonas paucimobilis UT26. Stereochemical analysis of the reaction products formed during conversion of g-HCH by LinA was investigated by GC-MS, NMR, CD and molecular modeling. The NMR spectra of 1,3,4,5,6-pentachlorocyclohexene (PCCH) produced from g-HCH using either enzymatic dehydrochlorination or alkaline dehydrochlorination were compared and found to be identical. Both enantiomers present in the racemate of synthetic g-PCCH were converted by LinA, each at a different rate. 1,2,4-trichlorobenzene (1,2,4-TCB) was detected as the only product of the biotransformation of biosynthetic g-PCCH. 1,2,4-TCB and 1,2,3-TCB were identified as the dehydrochlorination products of racemic g-PCCH. d-PCCH was detected as the only product of dehydrochlorination of d-HCH. LinA requires the presence of a 1,2-biaxial HCl pair on a substrate molecule. LinA enantiotopologically differentiates two 1,2-biaxial HCl pairs present on g-HCH and gives rise to a single PCCH enantiomer 1,3(R),4(S),5(S),6(R)-PCCH. Furthermore, LinA enantiomerically differentiates 1,3(S),4(R),5(R),6(S)-PCCH and 1,3(R),4(S),5(S),6(R)-PCCH. The proposed mechanism of enzymatic biotransformation of g-HCH to 1,2,4-TCB by LinA consists of two 1,2-anti conformationally dependent dehydrochlorinations followed by 1,4-anti dehydrochlorination.

Links

GA203/97/P149, research and development project

Name: Studium molekulárních mechanismů biodegradačních reakcí - konstrukce QSBR modelů a proteinové inženýrství dehalogenas

Investor: Czech Science Foundation, Study of the molecular mechanisms of biodegradation reactions - construction of QSBR models and protein engineering of haloalkane dehalogenases

LN00A016, research and development project

Name: BIOMOLEKULÁRNÍ CENTRUM

Investor: Ministry of Education, Youth and Sports of the CR, Biomolecular Center

ME 276, research and development project

Name: Racionální re-design mikrobiálních enzymů podílejících se na degradaci toxických organických polutantů

Investor: Ministry of Education, Youth and Sports of the CR, Rational re-design of microbial enzymes involved in degradation of toxic organic pollutants.

Cite

TRANTÍREK, Lukáš; Kamila HYNKOVÁ; Yuji NAGATA; Alexey MURZIN; Alena ANSORGOVÁ; Vladimír SKLENÁŘ and Jiří DAMBORSKÝ. Reaction mechanism and stereochemistry of gamma-hexachlorocyclohexane dehydrochlorinase LinA. Journal of Biological Chemistry. 2001, vol. 276, No 11, p. 7734-7740.

@article{360452,
   author = {Trantírek, Lukáš and Hynková, Kamila and Nagata, Yuji and Murzin, Alexey and Ansorgová, Alena and Sklenář, Vladimír and Damborský, Jiří},
   article_number = {11},
   language = {eng},
   journal = {Journal of Biological Chemistry},
   title = {Reaction mechanism and stereochemistry of gamma-hexachlorocyclohexane dehydrochlorinase LinA},
   url = {http://www.ncbr.chemi.muni.cz/~jiri/ABSTRACTS/jbc01.html},
   volume = {276},
   year = {2001}
}

TY  - JOUR
ID  - 360452
AU  - Trantírek, Lukáš - Hynková, Kamila - Nagata, Yuji - Murzin, Alexey - Ansorgová, Alena - Sklenář, Vladimír - Damborský, Jiří
PY  - 2001
TI  - Reaction mechanism and stereochemistry of gamma-hexachlorocyclohexane dehydrochlorinase LinA
JF  - Journal of Biological Chemistry
VL  - 276
IS  - 11
SP  - 7734
EP  - 7734
UR  - http://www.ncbr.chemi.muni.cz/~jiri/ABSTRACTS/jbc01.html
N2  - g-hexachlorocyclohexane dehydrochlorinase (LinA) catalyses the initial steps in the biotransformation of the important insecticide g-hexachlorocyclohexane (g-HCH) by the soil bacterium Sphingomonas paucimobilis UT26. Stereochemical analysis of the reaction products formed during conversion of g-HCH by LinA was investigated by GC-MS, NMR, CD and molecular modeling. The NMR spectra of 1,3,4,5,6-pentachlorocyclohexene (PCCH) produced from g-HCH using either enzymatic dehydrochlorination or alkaline dehydrochlorination were compared and found to be identical. Both enantiomers present in the racemate of synthetic g-PCCH were converted by LinA, each at a different rate. 1,2,4-trichlorobenzene (1,2,4-TCB) was detected as the only product of the biotransformation of biosynthetic g-PCCH. 1,2,4-TCB and 1,2,3-TCB were identified as the dehydrochlorination products of racemic g-PCCH. d-PCCH was detected as the only product of dehydrochlorination of d-HCH. LinA requires the presence of a 1,2-biaxial HCl pair on a substrate molecule. LinA enantiotopologically differentiates two 1,2-biaxial HCl pairs present on g-HCH and gives rise to a single PCCH enantiomer 1,3(R),4(S),5(S),6(R)-PCCH. Furthermore, LinA enantiomerically differentiates 1,3(S),4(R),5(R),6(S)-PCCH and 1,3(R),4(S),5(S),6(R)-PCCH. The proposed mechanism of enzymatic biotransformation of g-HCH to 1,2,4-TCB by LinA consists of two 1,2-anti conformationally dependent dehydrochlorinations followed by 1,4-anti dehydrochlorination.
ER  -

TRANTÍREK, Lukáš; Kamila HYNKOVÁ; Yuji NAGATA; Alexey MURZIN; Alena ANSORGOVÁ; Vladimír SKLENÁŘ and Jiří DAMBORSKÝ. Reaction mechanism and stereochemistry of gamma-hexachlorocyclohexane dehydrochlorinase LinA. \textit{Journal of Biological Chemistry}. 2001, vol.~276, No~11, p.~7734-7740.

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