Functional identification of a putative beta-galactosidase gene in the special lac gene cluster of Lactobacillus acidophilus.
The putative beta-galactosidase gene (lacZ) of Lactobacillus acidophilus has a very low degree of homology to the Escherichia coli beta-galactosidase gene (lacZ) and locates in a special lac gene cluster which contains two beta-galactosidase genes. No functional characteristic of the putative beta-galactosidase has been described so far. In this study, the lacZ gene of L. acidophilus was hetero-expressed in E. coli and the recombinant protein was purified by a three-step procedure. The product of the lacZ gene was also extracted from L. acidophilus ATCC 4356 and active staining was carried out. The enzymatic properties of the purified recombinant LacZ were assayed. The results of hetero-expression showed the recombinant LacZ without tag had beta-galactosidase activity. The purified recombinant LacZ had a specific activity of 43.2 U/mg protein. The result of active staining showed that the functional product of the lacZ gene did exist in L. acidophilus. The L. acidophilus beta-galactosidase (LacZ) had an optimal pH of 6, an optimal temperature of 37 degrees C and could hydrolyze 73% of lactose in milk in 30 h at 10 degrees C. The L. acidophilus beta-galactosidase (LacZ) was identified as cold-adapted beta-galactosidase in this study for the first time, and may be useful for lactose removal from dairy products at low temperatures.
Pan
Qu
Q
Department of Microbiology, Third Military Medical University, Chongqing, 400038, China.
Zhu
Junmin
J
Liu
Lina
L
Cong
Yanguang
Y
Hu
Fuquan
F
Li
Jinchuan
J
Yu
Xiaoping
X
D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D005822:Genetic Vectors; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007422:Intestines / Q000382:microbiology; D007700:Kinetics; D007779:Lactobacillus acidophilus / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D008970:Molecular Weight; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D001616:beta-Galactosidase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D017931:DNA Primers; D011994:Recombinant Proteins; D001616:beta-Galactosidase