Purification and characterization of a novel glucansucrase from Leuconostoc lactis EG001.
A gene encoding glucansucrase was identified in Leuconostoc lactis EG001 isolated from lactic acid bacteria (LAB) in Kimchi, a traditional Korean fermented food. The L. lactis EG001 glucansucrase gene consists of 4503 bp open reading frame (ORF) and encodes an enzyme of 1500 amino acids with an apparent molecular mass of 165 kDa. The deduced amino-acid sequence showed the highest amino-acid sequence identity (75%) to that of dextransucrase of L. mesenteroides. The gene was cloned and over-expressed in Escherichia coli strain. The recombinant enzyme was purified via Ni-NTA affinity chromatography and its enzymatic properties were characterized. The enzyme exhibited optimum activity at 30 degrees C and pH 5.0. In addition, the enzyme was able to catalyze the glycosylation of l-ascorbic acid to l-ascorbic acid 2-glucoside. The glycosylated product via EG001 glucansucrase has the potential as an antioxidant in industrial applications.
Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology, Jeonbuk 580-185, Republic of Korea.
Yong-Mo
YM
Kim
Min Ji
MJ
Yeon
Nack-Shick
NS
Choi
Young-Hyo
YH
Chang
Min Young
MY
Jung
Jae Jun
JJ
Song
Joong Su
JS
Kim
D000595:Amino Acid Sequence; D001483:Base Sequence; D016695:Glycosyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007934:Leuconostoc / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny
D016695:Glycosyltransferases; C033837:alternansucrase