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The cloning and characterization of a second α‐amylase of A. hydrophila JMP636
Author(s) -
Kidd S.P.,
Pemberton J.M.
Publication year - 2002
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2002.01529.x
Subject(s) - aeromonas hydrophila , amylase , biology , escherichia coli , alpha amylase , gene , enzyme , strain (injury) , cloning (programming) , biochemistry , genetics , bacteria , anatomy , computer science , programming language
Aims: The aim of this study was to identify, clone and characterize the second amylase of Aeromonas hydrophila JMP636, AmyB, and to compare it to AmyA. Methods and Results: The amylase activity of A. hydrophila JMP636 is encoded by multiple genes. A second genetically distinct amylase gene, amyB , has been cloned and expressed from its own promoter in Escherichia coli . AmyB is a large α‐amylase of 668 amino acids. Outside the conserved domains of α‐amylases there is limited sequence relationship between the two α‐amylases of A. hydrophila JMP636 AmyA and AmyB. Significant (80%) similarity exists between amyB and an α‐amylase of A. hydrophila strain MCC‐1. Differences in either the functional properties or activity under different environmental conditions as possible explanations for multiple copies of amylases in JMP636 is less likely after an examination of several physical properties, with each of the properties being very similar for both enzymes (optimal pH and temperature, heat instability). However the reaction end products and substrate specificity did vary enough to give a possible reason for the two enzymes being present. Both enzymes were confirmed to be α‐type amylases. Conclusions: AmyB has been isolated, characterized and then compared to AmyA. Significance and Impact of Study: The amylase phenotype is rarely encoded by more than one enzyme within one strain, this study therefore allows the better understanding of the unusual amylase production by A. hydrophila.