Cloning and Sequence Analysis of Vibrio halioticoli Genes Encoding Three Types of Polyguluronate Lyase

The alginate lyase-coding genes of Vibrio halioticoli IAM 14596(T), which was isolated from the gut of the abalone Haliotis discus hannai, were cloned using plasmid vector pUC 18, and expressed in Escherichia coli. Three alginate lyase-positive clones, pVHB, pVHC, and pVHE, were obtained, and all cl...

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Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2000-01, Vol.2 (1), p.65-73
Main Authors: Sugimura, I, I, Sawabe, T, Ezura, Y
Format: Article
Language:English
Subjects:
Alginate lyase
Alginic acid
alyVG1 gene
alyVG2 gene
alyVG3 gene
Amino acid sequences
Amino acids
Bacteria
Clones
Cloning
DNA
E coli
Enzymatic activity
Enzyme activity
Enzymes
Escherichia coli
Genes
Haliotis discus hannai
Homology
Klebsiella
Lyases
Marine
Marine molluscs
Nucleotides
Plasmids
polyguluronate lyase
Residues
Seaweed meal
Sequencing
Vibrio halioticoli
Waterborne diseases
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Summary:The alginate lyase-coding genes of Vibrio halioticoli IAM 14596(T), which was isolated from the gut of the abalone Haliotis discus hannai, were cloned using plasmid vector pUC 18, and expressed in Escherichia coli. Three alginate lyase-positive clones, pVHB, pVHC, and pVHE, were obtained, and all clones expressed the enzyme activity specific for polyguluronate. Three genes, alyVG1, alyVG2, and alyVG3, encoding polyguluronate lyase were sequenced: alyVG1 from pVHB was composed of a 1056-bp open reading frame (ORF) encoding 352 amino acid residues; alyVG2 gene from pVHC was composed of a 993-bp ORF encoding 331 amino acid residues; and alyVG3 gene from pVHE was composed of a 705-bp ORF encoding 235 amino acid residues. Comparison of nucleotide and deduced amino acid sequences among AlyVG1, AlyVG2, and AlyVG3 revealed low homologies. The identity value between AlyVG1 and AlyVG2 was 18.7%, and that between AlyVG2 and AlyVG3 was 17.0%. A higher identity value (26.0%) was observed between AlyVG1 and AlyVG3. Sequence comparison among known polyguluronate lyases including AlyVG1, AlyVG2, and AlyVG3 also did not reveal an identical region in these sequences. However, AlyVG1 showed the highest identity value (36.2%) and the highest similarity (73.3%) to AlyA from Klebsiella pneumoniae. A consensus region comprising nine amino acid (YFKAGXYXQ) in the carboxy-terminal region previously reported by Mallisard and colleagues was observed only in AlyVG1 and AlyVG2.
ISSN:1436-2228
1436-2236
DOI:10.1007/s101269900010