Purification and characterisation of the BIOH protein from the biotin biosynthetic pathway

FEBS Letters

Campopiano, Dominic J¹  Crichton, Hilary J¹  Nettleship, Joanne E¹  Tomczyk, Nicholas H¹  Baxter, Robert L¹  Webster, Scott P¹ 

[1] Department of Chemistry, Joseph Black Building, University of Edinburgh, West Mains Road, Edinburgh EH9 3JJ, UK

关键词: Pimeloyl-coenzyme A;    Biotin;    Biosynthesis;    Coenzyme A;    Mass spectrometry;    CoA;    coenzyme A;    PP;    phosphopantetheine;    ACP;    acyl carrier protein;    ACPS;    acyl carrier protein synthase;    DTT;    dithiothreitol;    PCR;    polymerase chain reaction;    LC-MS;    liquid chromatography-mass spectrometry;    AONS;    8-amino-7-oxononanoate synthase;    DANS;    7;    8-diaminononanoate synthase;    DTBS;    dethiobiotin synthetase;    BS;    biotin synthase;   

DOI  :  10.1016/S0014-5793(02)02342-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Conversion of pimeloyl-coenzyme A (CoA) to biotin in Escherichia coli requires at least four enzymes encoded by genes in the bio operon. One gene, bioH, which is not present in the bioABFCD operon, is required for the synthesis of pimeloyl-CoA but its exact role in formation of this intermediate is unknown. To investigate this further, we have overexpressed and purified the bioH gene products from both E. coli (BIOH EC) and Neisseria meningitis (BIOH NM) in E. coli. When purified BIOH was incubated with excess CoA and analysed by electrospray mass spectrometry a species of mass corresponding to a BIOH:CoA complex was observed. Mutation of a conserved serine residue to alanine (BIOH EC S82A) did not prevent CoA binding. This is the first report of the purification of BIOH and the observation of a small molecule bound to the protein provides clues to its role in pimeloyl-CoA synthesis.

【 授权许可】

Unknown