Resolution and some properties of enzymes involved in enantioselective transformation of 1,3-dichloro-2-propanol to (R)-3-chloro-1,2-propanediol by Corynebacterium sp. strain N-1074

Nakamura, T.; Nagasawa, Tôru; Yu, Fujio; Watanabe, Ichirô; Yamada, Hideaki

doi:10.1128/jb.174.23.7613-7619.1992

Cited by 66 publications

(35 citation statements)

References 34 publications

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“…The substrate range of recombinant HheA AD2 was in agreement with that of the enzyme isolated from strain AD2 (31). The substrate range of HheB GP1 was similar to those reported for HheB (20,21) (Table 2) and DehA (2). Halohydrin dehalogenase HheC displayed a relatively high level of activity with chloroacetone, which clearly distinguishes this enzyme from HheA AD2 and HheB GP1 .…”

Section: Resultssupporting

confidence: 73%

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Halohydrin Dehalogenases Are Structurally and Mechanistically Related to Short-Chain Dehydrogenases/Reductases

et al. 2001

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Halohydrin dehalogenases, also known as haloalcohol dehalogenases or halohydrin hydrogen-halide lyases, catalyze the nucleophilic displacement of a halogen by a vicinal hydroxyl function in halohydrins to yield epoxides. Three novel bacterial genes encoding halohydrin dehalogenases were cloned and expressed in Escherichia coli, and the enzymes were shown to display remarkable differences in substrate specificity. The halohydrin dehalogenase of Agrobacterium radiobacter strain AD1, designated HheC, was purified to homogeneity. The k cat and K m values of this 28-kDa protein with 1,3-dichloro-2-propanol were 37 s ؊1 and 0.010 mM, respectively. A sequence homology search as well as secondary and tertiary structure predictions indicated that the halohydrin dehalogenases are structurally similar to proteins belonging to the family of short-chain dehydrogenases/reductases (SDRs). Moreover, catalytically important serine and tyrosine residues that are highly conserved in the SDR family are also present in HheC and other halohydrin dehalogenases. The third essential catalytic residue in the SDR family, a lysine, is replaced by an arginine in halohydrin dehalogenases. A site-directed mutagenesis study, with HheC as a model enzyme, supports a mechanism for halohydrin dehalogenases in which the conserved Tyr145 acts as a catalytic base and Ser132 is involved in substrate binding. The primary role of Arg149 may be lowering of the pK a of Tyr145, which abstracts a proton from the substrate hydroxyl group to increase its nucleophilicity for displacement of the neighboring halide. The proposed mechanism is fundamentally different from that of the well-studied hydrolytic dehalogenases, since it does not involve a covalent enzyme-substrate intermediate.Halogenated aliphatics constitute an important class of environmental pollutants. Various microorganisms have evolved that are able to degrade some of these compounds and use them as sole sources of carbon and energy. Such organisms are of importance for bioremediation of polluted soil, groundwater, and wastewater. In most cases, specialized enzymes, designated dehalogenases, catalyze the cleavage of the carbonhalogen bonds, which is a key detoxification reaction. Hydrolytic dehalogenases have been studied extensively, which has resulted in detailed insight into the structure and mechanism of several enzymes of this class (8,33). For other dehalogenases, structural and mechanistic data are hardly available.Halohydrin dehalogenases, also referred to as haloalcohol dehalogenases or halohydrin hydrogen-halide lyases, occur in the degradation pathways of halopropanols and 1,2-dibromoethane, where they catalyze the nucleophilic displacement of a halogen by a vicinal hydroxyl group in halohydrins, yielding an epoxide, a proton, and a halide ion (7,22,30,31). These enzymes also efficiently catalyze the reverse reaction, the halogenation of epoxides, and the dehalogenation of vicinal chlorocarbonyls to hydroxycarbonyls (2, 14, 31). The interest in halohydrin dehalogenases increased when i...

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Section: Resultssupporting

confidence: 73%

“…The specific activity of HheC with this substrate is similar to that reported for HheA AD2 (31). However, the K m value of HheC for 1,3-dichloro-2-propanol is 2 to 3 orders of magnitude lower than those of HheA, HheA AD2 , and HheB (17,21,31).…”

Section: Resultssupporting

confidence: 48%

Halohydrin Dehalogenases Are Structurally and Mechanistically Related to Short-Chain Dehydrogenases/Reductases

et al. 2001

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“…epoxide hydrolase. This might be the carboxyl terminus of another epoxide hydrolase 4 ) of Corynebacterium sp. strain N-I074, although Jacobs et al mentioned that the other peptide (fragment 2 in ref.…”

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confidence: 99%

“…strain N-I074 into two OCP-dechlorinating activities and two ECH-hydrolyzing activities. 4 ) We report here cloning of the hheA and hheB genes encoding two isoenzymes involved in the first step reaction from Corynebacterium sp. strain N-I074 and their nucleotide sequence and expression in E. coli.…”

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confidence: 99%

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Cloning of Two Halohydrin Hydrogen–Halide–Lyase Genes ofCorynebacteriumsp. Strain N-1074 and Structural Comparison of the Genes and Gene Products

Nakamura

Mizunashi

et al. 1994

Bioscience, Biotechnology, and Biochemistry

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Optically active chlorohydrins as chiral C3 and C4 building units: Microbial resolution and synthetic applications

1998

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Production of highly optically active C3 and C4 chlorohydrins was developed by using the bacteria stereoselectively dehalogenating and assimilating the racemic substrate: Pseudomonas sp. and Alcaligenes sp. These bacteria stereoselectively assimilate (RS)‐2,3‐dichloro‐1‐propanol (DCP) and (RS)‐3‐chloro‐1,2‐propanediol (CPD) followed by microbial preparation of (R)‐ and (S)‐DCP, and (R)‐ and (S)‐CPD with >99% ee. A novel dehalogenating enzyme, halohydrin dehydro‐dehalogenase from one of the above strains, Alcaligenes sp. DS‐S‐7G, was applicable for preparation of optically active 1,2‐diols with 60–99% ee. Moreover, microbial resolution of C4 chlorohydrins with whole cells of Pseudomonas sp. was carried out. This resolution reaction using the resting cells gave (R)‐ and (S)‐4‐chloro‐3‐hydroxybutyrate (CHB) and (S)‐4‐chloro‐3‐hydroxybutyronitrile (BN) with >98% ee. In the case of the resting cells of Enterobacter sp., both (R)‐CHB (>99% ee) and (S)‐3‐hydroxy‐γ‐butyrolactone (95% ee) with excellent yield were obtained. Also, some typical synthetic applications using the above chiral C3 and C4 synthons were introduced: ferroelectric liquid crystals, optically active β‐blockers, and other chiral pharmaceuticals. Chirality 10:682–692, 1998. © 1998 Wiley‐Liss, Inc.

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Resolution and some properties of enzymes involved in enantioselective transformation of 1,3-dichloro-2-propanol to (R)-3-chloro-1,2-propanediol by Corynebacterium sp. strain N-1074

Abstract: During the course of the transformation of 1,3-dichloro-2-propanol (DCP) into (R)-3-chloro-1,2-propanediol

Cited by 66 publications

References 34 publications

Halohydrin Dehalogenases Are Structurally and Mechanistically Related to Short-Chain Dehydrogenases/Reductases

Halohydrin Dehalogenases Are Structurally and Mechanistically Related to Short-Chain Dehydrogenases/Reductases

Cloning of Two Halohydrin Hydrogen–Halide–Lyase Genes ofCorynebacteriumsp. Strain N-1074 and Structural Comparison of the Genes and Gene Products

Optically active chlorohydrins as chiral C3 and C4 building units: Microbial resolution and synthetic applications

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