Biochemical properties and substrate specificities of a recombinantly produced Azotobacter vinelandii alginate lyase
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Author(s)
Ertesvag, H
Erlien, F
Skjak-Braek, G
Rehm, BHA
Valla, S
Griffith University Author(s)
Year published
1998
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Alginate is a polysaccharide composed of β-D-mannuronic acid (M) and α-L-guluronic acid (G). An Azotobacter vinelandii alginate lyase gene,algL, was cloned, sequenced, and expressed inEscherichia coli. The deduced molecular mass of the corresponding protein is 41.4 kDa, but a signal peptide is cleaved off, leaving a mature protein of 39 kDa. Sixty-three percent of the amino acids in this mature protein are identical to those in AlgL fromPseudomonas aeruginosa. AlgL was partially purified, and the activity was found to be optimal at a pH of 8.1 to 8.4 and at 0.35 M NaCl. Divalent cations are not necessary for activity. The ...
View more >Alginate is a polysaccharide composed of β-D-mannuronic acid (M) and α-L-guluronic acid (G). An Azotobacter vinelandii alginate lyase gene,algL, was cloned, sequenced, and expressed inEscherichia coli. The deduced molecular mass of the corresponding protein is 41.4 kDa, but a signal peptide is cleaved off, leaving a mature protein of 39 kDa. Sixty-three percent of the amino acids in this mature protein are identical to those in AlgL fromPseudomonas aeruginosa. AlgL was partially purified, and the activity was found to be optimal at a pH of 8.1 to 8.4 and at 0.35 M NaCl. Divalent cations are not necessary for activity. The pI of the enzyme is 5.1. When an alginate rich in mannuronic acid was used as the substrate, the Km was found to be 4.6 × 10−4 M (sugar residues). AlgL was found to cleave M-M and M-G bonds but not G-M or G-G bonds. Bonds involving acetylated residues were also cleaved, but this activity may be sensitive to the extent of acetylation.
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View more >Alginate is a polysaccharide composed of β-D-mannuronic acid (M) and α-L-guluronic acid (G). An Azotobacter vinelandii alginate lyase gene,algL, was cloned, sequenced, and expressed inEscherichia coli. The deduced molecular mass of the corresponding protein is 41.4 kDa, but a signal peptide is cleaved off, leaving a mature protein of 39 kDa. Sixty-three percent of the amino acids in this mature protein are identical to those in AlgL fromPseudomonas aeruginosa. AlgL was partially purified, and the activity was found to be optimal at a pH of 8.1 to 8.4 and at 0.35 M NaCl. Divalent cations are not necessary for activity. The pI of the enzyme is 5.1. When an alginate rich in mannuronic acid was used as the substrate, the Km was found to be 4.6 × 10−4 M (sugar residues). AlgL was found to cleave M-M and M-G bonds but not G-M or G-G bonds. Bonds involving acetylated residues were also cleaved, but this activity may be sensitive to the extent of acetylation.
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Journal Title
Journal of Bacteriology
Volume
180
Issue
15
Publisher URI
Copyright Statement
© 1998 American Society for Microbiology. The attached file is reproduced here in accordance with the copyright policy of the publisher. Please refer to the journal's website for access to the definitive, published version.
Subject
Biological sciences
Biochemistry and cell biology not elsewhere classified
Agricultural, veterinary and food sciences
Biomedical and clinical sciences