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Pubmed ID :16326697
Publication Date : //

Cloning, expression, and characterization of bacterial L-arabinose 1-dehydrogenase involved in an alternative pathway of L-arabinose metabolism.


Azospirillum brasiliense converts L-arabinose to alpha-ketoglutarate via five hypothetical enzymatic steps. We purified and characterized L-arabinose 1-dehydrogenase (EC 1.1.1.46), catalyzing the conversion of L-arabinose to L-arabino-gamma-lactone as an enzyme responsible for the first step of this alternative pathway of L-arabinose metabolism. The purified enzyme preferred NADP+ to NAD+ as a coenzyme. Kinetic analysis revealed that the enzyme had high catalytic efficiency for both L-arabinose and D-galactose. The gene encoding L-arabinose 1-dehydrogenase was cloned using a partial peptide sequence of the purified enzyme and was overexpressed in Escherichia coli as a fully active enzyme. The enzyme consists of 308 amino acids and has a calculated molecular mass of 33,663.92 Da. The deduced amino acid sequence had some similarity to glucose-fructose oxidoreductase, D-xylose 1-dehydrogenase, and D-galactose 1-dehydrogenase. Site-directed mutagenesis revealed that the enzyme possesses unique catalytic amino acid residues. Northern blot analysis showed that this gene was induced by L-arabinose but not by D-galactose. Furthermore, a disruptant of the L-arabinose 1-dehydrogenase gene did not grow on L-arabinose but grew on D-galactose at the same growth rate as the wild-type strain. There was a partial gene for L-arabinose transport in the flanking region of the L-arabinose 1-dehydrogenase gene. These results indicated that the enzyme is involved in the metabolism of L-arabinose but not D-galactose. This is the first identification of a gene involved in an alternative pathway of L-arabinose metabolism in bacterium.

Authors : Watanabe Seiya , Kodaki Tsutomu , Kodak Tsutomu , Makino Keisuke ,

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Catalog number Product name Quantity
B673 Phenol Red Arabinose Broth USE For Arabinose fermentation studies of microorganisms. Qty per Litre of Medium: 21
B673 Phenol Red Arabinose Broth USE : For Arabinose fermentation studies of microorganisms. 5x500gm
ARA-021 L-[1-13C]arabinose CAS: 10 mg
ARA-002 D-[2-13C]arabinose CAS: 10 mg
ARA-001 D-[1-13C]arabinose CAS: 25 mg
087 D(_) ARABINOSE, 25 gm
ARA-003 D-[3-13C]arabinose CAS: 2.5 mg
ARA-007 D-[1-2H]arabinose CAS: 10 mg
259 L (+) ARABINOSE, 25 gm
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ARA-008 D-[2-2H]arabinose CAS: 10 mg
ARA-010 L-[2-13C]arabinose CAS: 10 mg
ARA-009 D-[5,5'-2H2]arabinose CAS: 10 mg
087 D(_) ARABINOSE, 10 gm
259 L (+) ARABINOSE, 10 gm
259 L (+) ARABINOSE, 25 gm
087 D(_) ARABINOSE, 10 gm
087 D(_) ARABINOSE, 25 gm
ARA-004 D-[5-13C]arabinose CAS: 10 mg
ARA-019 D-[2,3,4,5-13C4]arabinose CAS: 2.5 mg
A35070 L-Arabinose 100 G
AB0071D D-Arabinose 25g
03330-74 D‐(-)‐Arabinose 5G
03330-32 D‐(-)‐Arabinose 25G
03306-04 L‐(+)‐Arabinose 5G


 

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