Open AccessPurification and characterization of the recombinant alginate lyase from Pseudomonas sp. leaked by Escherichia coli upon addition of glycine
Author(s) -
Fujiyama Kazuhito,
Maki Hideaki,
Kinoshita Shinichi,
Yoshida Toshiomi
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07384.x
Subject(s) - escherichia coli , extracellular , biochemistry , pseudomonas , glycine , biology , recombinant dna , peptide sequence , enterobacteriaceae , lyase , microbiology and biotechnology , pseudomonadaceae , plasmid , amino acid , enzyme , bacteria , gene , genetics
The plasmid pAL205 encodes an alginate lyase gene of Pseudomonas sp. OS‐ALG‐9, fused in frame to the β‐galactosidase α‐peptide gene. The alginate lyase (Aly) expressed in Escherichia coli (pAL205) was significantly secreted into the medium by the addition of glycine. The extracellular enzyme isolated from the culture of E. coli JM109 (pAL205) was purified over 15 000‐fold by successive chromatography and subjected to amino acid sequence analysis. The sequence determined was identical to that of the intracellular protein. Since the activity and molecular size of the extracellular Aly is identical to the intracellular protein and to the Aly isolated from Pseudomonas , the glycine does not affect or modify the Aly during its leakage into the medium.