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Organization and characterization of three genes involved in D-xylose catabolism in Lactobacillus pentosus

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Author: Lokman, B.C. · Santen, P. van · Verdoes, J.C. · Kruse, J. · Leer, R.J. · Posno, M. · Pouwels, P.H.
Type:article
Date:1991
Institution: Medisch Biologisch Laboratorium TNO
Source:Molecular and General Genetics, 1-2, 230, 161-169
Identifier: 231696
doi: doi:10.1007/BF00290664
Keywords: Biology · Amino acid comparison · D-xylose isomerase · D-xylulose kinase · NMR analysis · Regulatory protein · Xylose · Xylose isomerase · Amino acid sequence · Gene structure · Lactobacillus · Nonhuman · Nucleotide sequence · Priority journal · Sequence homology · Amino Acid Sequence · Bacillus subtilis · Base Sequence · Carbohydrate Epimerases · Cloning, Molecular · DNA, Bacterial · Genes, Bacterial · Lactobacillus · Magnetic Resonance Spectroscopy · Molecular Sequence Data · Multigene Family · Open Reading Frames · Phosphotransferases · Plasmids · Repressor Proteins · Restriction Mapping · Sequence Alignment · Sequence Homology, Nucleic Acid · Streptomyces · Xylose

Abstract

A cluster of three genes involved in D-xylose catabolism (viz. xylose genes) in Lactobacillus pentosus has been cloned in Escherichia coli and characterized by nucleotide sequence analysis. The deduced gene products show considerable sequence similarity to a repressor protein involved in the regulation of expression of xylose genes in Bacillus subtilis (58%), to E. coli and B. subtilis D-xylose isomerase (68% and 77%, respectively), and to E. coli D-xylulose kinase (58%). The cloned genes represent functional xylose genes since they are able to complement the inability of a L. casei strain to ferment D-xylose. NMR analysis confirmed that 13C-xylose was converted into 13C-acetate in L. casei cells transformed with L. pentosus xylose genes but not in untransformed L. casei cells. Comparison with the aligned amino acid sequences of D-xylose isomerases of different bacteria suggests that L. pentosus D-xylose isomerase belongs to the same similarity group as B. subtilis and E. coli D-xylose isomerase and not to a second similarity group comprising D-xylose isomerases of Streptomyces violaceoniger, Ampullariella sp. and Actinoplanes. The organization of the L. pentosus xylose genes, 5'-xylR (1167 bp, repressor) - xylA (1350 bp, D-xylose isomerase) - xylB (1506 bp, D-xylulose kinase) - 3' is similar to that in B. subtilis. In contrast to B. subtilis xylR, L. pentosus xylR is transcribed in the same direction as xylA and xylB. Molecular Sequence Numbers: GENBANK: M57384, S55527, S55528, S55529, S55530, S55531, S55532, S63909, S69823, Z14057; Chemicals/CAS: Carbohydrate Epimerases, EC 5.1.3; DNA, Bacterial; Phosphotransferases, EC 2.7; Plasmids; Repressor Proteins; xylose isomerase, EC 5.3.1.5; Xylose; xylulokinase, EC 2.7.1.17