RT Journal Article SR Electronic(1) A1 Maki, Hideaki A1 Mori, Atsutoshi A1 Fujiyama, Kazuhito A1 Kinoshita, Shinichi A1 Yoshida, ToshiomiYR 1993 T1 Cloning, sequence analysis and expression in Escherichia coli of a gene encoding an alginate lyase from Pseudomonas sp. OS-ALG-9 JF Microbiology, VO 139 IS 5 SP 987 OP 993 DO https://doi.org/10.1099/00221287-139-5-987 PB Microbiology Society, SN 1465-2080, AB Summary: 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 I365 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., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-139-5-987