Document Detail


Enzymatic properties of futalosine hydrolase, an enzyme essential to a newly identified menaquinone biosynthetic pathway.
MedLine Citation:
PMID:  19420717     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In prokaryotes, menaquinone is used for respiration. In Escherichia coli, menaquinone is biosynthesized from chorismate by seven enzymes. However, very recently, we identified an alternative pathway (the futalosine pathway), which operates in some bacteria, including Streptomyces coelicolor, Helicobacter pylori, Campylobacter jejuni, and Thermus thermophilus. We describe the steps of this pathway, which branches at chorismate in a manner similar to the known pathway, but then follows a different route. This new pathway includes futalosine, an unusual nucleoside derivative consisting of inosine and o-substituted benzoate moieties, as a biosynthetic intermediate. In this study, a recombinant futalosine hydrolase (TTHA0556) of T. thermophilus, which participates in the second step of the pathway and catalyzes the reaction releasing hypoxanthine from futalosine, was prepared and used in functional analyses. Recombinant TTHA0556 formed a homotetramer and reacted only with futalosine; other structurally related nucleotides and nucleosides were not accepted. Recombinant TTHA0556 required no cofactors, and the optimum pH and temperature were 4.5 and 80 degrees C. The Km value was calculated to be 154.0+/-5.3 microM and the kcat value was 1.02/s. Recombinant TTHA0556 was slightly inhibited by hypoxanthine, with a Ki value of 1.1 mM.
Authors:
Tomoshige Hiratsuka; Nobuya Itoh; Haruo Seto; Tohru Dairi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-05-07
Journal Detail:
Title:  Bioscience, biotechnology, and biochemistry     Volume:  73     ISSN:  1347-6947     ISO Abbreviation:  Biosci. Biotechnol. Biochem.     Publication Date:  2009 May 
Date Detail:
Created Date:  2009-05-25     Completed Date:  2009-08-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9205717     Medline TA:  Biosci Biotechnol Biochem     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  1137-41     Citation Subset:  IM    
Affiliation:
Biotechnology Research Center, Toyama Prefectural University, Toyama, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
Hydrogen-Ion Concentration
Hydrolases / antagonists & inhibitors,  chemistry,  metabolism*
Hypoxanthine / pharmacology
Kinetics
Metals / pharmacology
Nucleosides / metabolism*
Protein Structure, Quaternary
Recombinant Fusion Proteins / antagonists & inhibitors,  chemistry,  metabolism
Substrate Specificity
Temperature
Thermus thermophilus / enzymology*,  metabolism
Vitamin K 2 / metabolism*
Chemical
Reg. No./Substance:
0/Metals; 0/Nucleosides; 0/Recombinant Fusion Proteins; 0/futalosine; 11032-49-8/Vitamin K 2; 68-94-0/Hypoxanthine; EC 3.-/Hydrolases

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