Cloning, expression and characterization of a gene encoding nitroalkane-oxidizing enzyme from Streptomyces ansochromogenes

Zhang, Jihui, Ma, Wenbo and Tan, Huarong (2002) Cloning, expression and characterization of a gene encoding nitroalkane-oxidizing enzyme from Streptomyces ansochromogenes. European Journal of Biochemistry, 269 (24). pp. 6302-6307. ISSN 0014-2956

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Abstract

A nitroalkane-oxidizing enzyme gene (naoA) was cloned from a genomic DNA library of Streptomyces ansochromogenes 7100. The deduced protein (NaoA) of this gene contains 363 amino acids and has high similarity to several nitroalkane-oxidizing enzymes from various micro-organisms. The naoA gene was subcloned into an expression vector pET23b and overexpressed in Escherichia coli BL21(DE3). The protein was then purified, and its characteristics were studied. Experimental results showed that NaoA can convert 1-nitropropane, 2-nitropropane and nitroethane into the corresponding carbonyl compounds. The optimal pH and temperature for NaoA was found to be pH 7-8 and 48-56°C, respectively. The Km of NaoA for nitroethane is ≈26.8 mM. NADH and nitro blue tetrazolium are strong inhibitors of NaoA, and thiol compounds and superoxide dismutase partially inhibit the enzyme activity. Therefore, superoxide may be an essential intermediate in the oxidation of nitroalkane by NaoA.

Item Type: Article
Uncontrolled Keywords: enzymatic properties,expression,gene cloning,nitroalkane-oxidizing enzyme,streptomyces,biochemistry ,/dk/atira/pure/subjectarea/asjc/1300/1303
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Depositing User: LivePure Connector
Date Deposited: 12 Sep 2022 12:31
Last Modified: 30 Sep 2022 02:30
URI: https://ueaeprints.uea.ac.uk/id/eprint/88183
DOI: 10.1046/j.1432-1033.2002.03350.x

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