The Nitrile‐Forming Enzyme 7‐Cyano‐7‐Deazaguanine Synthase from Geobacillus kaustophilus: A Reverse Nitrilase?. Issue 16 (14th October 2015)

• Record Type: Journal Article
• Authors: Winkler, Margit; Dokulil, Katharina; Weber, Hansjörg; Pavkov‐Keller, Tea; Wilding, Birgit
• Is Part Of: Chembiochem. Volume 16:Issue 16(2015)
• Page Start: 2373
• Page End: 2378
• Publication Date: 2015-10-14
• Subjects: Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201500335
• Languages: English
• ISSNs: 1439-4227

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>7‐Cyano‐7‐deazaguanine synthase (E.C. 6.3.4.20) is an enzyme that catalyzes the formation of a nitrile from a carboxylic acid and ammonia at the expense of ATP. The protein from <italic>G. kaustophilus</italic> was heterologously expressed, and its biochemical characteristics were explored by using a newly developed HPLC‐MS based assay, ³¹P NMR, and a fluorescence‐based thermal‐shift assay. The protein showed the expected high thermostability, had a pH optimum at pH 9.5, and an apparent temperature optimum at 60 °C. We observed strict substrate specificity of QueC for the natural substrate 7‐carboxy‐7‐deazaguanine, and determined AMP and pyrophosphate as co‐products of preQ₀.</p> </abstract>