Cloning and biochemical characterization of a glucosidase from a marine bacterium Aeromonas sp. HC11e-3.
By constructing the genomic library, a β-glucosidase gene, with a length of 2,382 bp, encoding 793 amino acids, designated bgla, is cloned from a marine bacterium Aeromonas sp. HC11e-3. The enzyme is expressed successfully in the recombinant host Escherichia coli BL21 (DE3) and purified using glutathione affinity purification system. It shows the optimal activity at pH 6, 55 °C and hydrolyzes aryl-glucoside specially. Ca(2+), Mn(2+), Zn(2+), Ba(2+), Pb(2+), Sr(2+) can activate the enzyme activity, whereas SDS, EDTA, DTT show slight inhibition to the enzyme activity. Homologous comparing shows that the enzyme belongs to glycosyl hydrolase family 3, exhibiting 46 % identity with a fully characterized glucosidase from Thermotoga neapolitana DSM 4359. Such results provide useful references for investigating other glucosidases in the glycosyl family 3 as well as developing glucosidases using in suitable industrial area.
State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
Xiaoluo
X
Huang
Yan
Y
Zhao
Yunjing
Y
Dai
Gaobing
G
Wu
Zongze
Z
Shao
Qinglan
Q
Zeng
Ziduo
Z
Liu
D000333:Aeromonas / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D002413:Cations, Divalent / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D020536:Enzyme Activators / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005959:Glucosidases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012623:Seawater / Q000382:microbiology; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D046949:Thermotoga neapolitana / Q000201:enzymology / Q000235:genetics
D002413:Cations, Divalent; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D008670:Metals; D011994:Recombinant Proteins; D005959:Glucosidases