title: Cloning, sequence, and expression of a chitinase gene from a marine bacterium, Altermonas sp. strain O-7.
abstract: The gene encoding an extracellular chitinase from marine Alteromonas sp. strain O-7 was cloned in Escherichia coli JM109 by using pUC18. The chitinase produced was not secreted into the growth medium but accumulated in the periplasmic space. A chitinase-positive clone of E. coli produced two chitinases with different molecular weights from a single chitinase gene. These proteins showed almost the same enzymatic properties as the native chitinase of Alteromonas sp. strain O-7. The N-terminal sequences of the two enzymes were identical. The nucleotide sequence of the 3,394-bp SphI-HindIII fragment that included the chitinase gene was determined. A single open reading frame was found to encode a protein consisting of 820 amino acids with a molecular weight of 87,341. A putative ribosome-binding site, promoter, and signal sequence were identified. The deduced amino acid sequence of the cloned chitinase showed sequence homology with chitinases A (33.4%) and B (15.3%) from Serratia marcescens. Regardless of origin, the enzymes of the two bacteria isolated from marine and terrestrial environments had high homology, suggesting that these organisms evolved from a common ancestor.
authors.affiliation: Osaka University of Pharmaceutical Sciences, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Tsujibo
authors.forename: H
authors.initials: H
authors.lastname: Orikoshi
authors.forename: H
authors.initials: H
authors.lastname: Tanno
authors.forename: K
authors.initials: K
authors.lastname: Fujimoto
authors.forename: K
authors.initials: K
authors.lastname: Miyamoto
authors.forename: C
authors.initials: C
authors.lastname: Imada
authors.forename: Y
authors.initials: Y
authors.lastname: Okami
authors.forename: Y
authors.initials: Y
authors.lastname: Inamori
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002688:Chitinases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D005798:Genes, Bacterial / Q000235:genetics*; D006088:Gram-Negative Aerobic Bacteria / Q000201:enzymology / Q000235:genetics*; D008386:Marine Biology; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
chemical_list: D011994:Recombinant Proteins; D002688:Chitinases