Cloning, sequence analysis and expression in Escherichia coli of a gene encoding an alginate lyase from Pseudomonas sp. OS-ALG-9
- PMID: 8336113
- DOI: 10.1099/00221287-139-5-987
Cloning, sequence analysis and expression in Escherichia coli of a gene encoding an alginate lyase from Pseudomonas sp. OS-ALG-9
Abstract
A gene (aly) encoding alginate lyase (ALY; EC 4.2.2.3) was isolated from a library constructed with the cosmid vector pHC79 and Sau3AI-digested genomic DNA of Pseudomonas sp. OS-ALG-9. Successive subcloning of the aly-containing cosmid enabled us to locate the gene on a 2.3 kb HpaI fragment. Nucleotide sequencing of this fragment revealed a single open reading frame (ORF) of 1365 bp. The directly determined N-terminal amino acid sequence of the ALY protein purified from Pseudomonas sp. OS-ALG-9 was found in the amino acid sequence deduced from this ORF between nucleotides 282 and 366. Expression of aly was induced by IPTG in Escherichia coli and leakage of the enzyme into the extracellular milieu was significantly enhanced by addition of glycine to the growth medium. The ALY enzyme had a greater specificity for the homopolymer of mannuronate than for that of guluronate.
Similar articles
-
Cloning and sequence analysis of the gene (alyII) coding for an alginate lyase of Pseudomonas sp. OS-ALG-9.Biosci Biotechnol Biochem. 1997 Nov;61(11):1853-7. doi: 10.1271/bbb.61.1853. Biosci Biotechnol Biochem. 1997. PMID: 9404064
-
Purification and characterization of the recombinant alginate lyase from Pseudomonas sp. leaked by Escherichia coli upon addition of glycine.FEMS Microbiol Lett. 1995 Feb 1;126(1):19-23. doi: 10.1111/j.1574-6968.1995.tb07384.x. FEMS Microbiol Lett. 1995. PMID: 7896072
-
High gene expression in Escherichia coli of recombinant alginate lyase as a fused protein with beta-galactosidase alpha-peptide.FEMS Microbiol Lett. 1995 Feb 1;126(1):13-7. doi: 10.1111/j.1574-6968.1995.tb07383.x. FEMS Microbiol Lett. 1995. PMID: 7896071
-
Polysaccharide lyase: molecular cloning of gellan lyase gene and formation of the lyase from a huge precursor protein in Bacillus sp. GL1.Arch Biochem Biophys. 1998 Jun 1;354(1):31-9. doi: 10.1006/abbi.1998.0674. Arch Biochem Biophys. 1998. PMID: 9633595
-
Vector design and development of host systems for Pseudomonas.Genet Eng (N Y). 2001;23:69-81. doi: 10.1007/0-306-47572-3_5. Genet Eng (N Y). 2001. PMID: 11570107 Review. No abstract available.
Cited by
-
Isolation and expression in Escherichia coli of cslA and cslB, genes coding for the chondroitin sulfate-degrading enzymes chondroitinase AC and chondroitinase B, respectively, from Flavobacterium heparinum.Appl Environ Microbiol. 2000 Jan;66(1):29-35. doi: 10.1128/AEM.66.1.29-35.2000. Appl Environ Microbiol. 2000. PMID: 10618199 Free PMC article.
-
Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms.Sci Rep. 2020 Jun 10;10(1):9390. doi: 10.1038/s41598-020-66293-2. Sci Rep. 2020. PMID: 32523130 Free PMC article.
-
Characterization of alginate lyase gene using a metagenomic library constructed from the gut microflora of abalone.J Ind Microbiol Biotechnol. 2012 Apr;39(4):585-93. doi: 10.1007/s10295-011-1054-0. Epub 2011 Nov 10. J Ind Microbiol Biotechnol. 2012. PMID: 22072436
-
Diversity of Three-Dimensional Structures and Catalytic Mechanisms of Alginate Lyases.Appl Environ Microbiol. 2018 Jan 17;84(3):e02040-17. doi: 10.1128/AEM.02040-17. Print 2018 Feb 1. Appl Environ Microbiol. 2018. PMID: 29150496 Free PMC article. Review.
-
Structural and functional aspects of mannuronic acid-specific PL6 alginate lyase from the human gut microbe Bacteroides cellulosilyticus.J Biol Chem. 2019 Nov 22;294(47):17915-17930. doi: 10.1074/jbc.RA119.010206. Epub 2019 Sep 17. J Biol Chem. 2019. PMID: 31530640 Free PMC article.
Publication types
MeSH terms
Substances
Associated data
- Actions