|
KMID : 0545120000100040559
|
|
Journal of Microbiology and Biotechnology
2000 Volume.10 No. 4 p.559 ~ p.563
|
|
|
Cloning and Sequencing of the α-1->6 Dextransucrase Gene from Leuconostoc mensenteroides B-742CB
|
|
Kim, Ho Sang
Kim, Do Man/Ryu, Hwa Ja/John F.Robyt
|
|
|
Abstract
|
|
|
|
A dextransucrase gene (dsrB742) that expresses a dextransucrase to synthesize mostly α-1→6 linked dextran with a low amount (3-5%) of α-1→3 branching was cloned and sequenced from Leuconostoc mesenteroides B-742CB. The 6.1-kb PstI fragments were ligated with pGEM-3Zf(-) and transformed into E. coli DH5α. The recombinant clone (pDSRB742) synthesized dextran on an agar plate containing 2%(w/v) sucrose. The dextran synthesized was hydrolyzed with Penicillium endo-dextranase. The hydrolyzate was composed of glucose, isomaltose, isomaltotriose, and branched pentasaccharide. The nucleotide sequence of dsrB742 showed one open reading frame (ORF) composed of 4,524bp encoding dextransucrase. The deduced amino acid sequence revealed a calculated molecular mass of 168.6kDa. It also showed an activity band of 184kDa on a non-denaturing SDS-PAGE (10%). The amino acid sequence of DSRB742 exhibited a 50% similarity with DSRA from L. mesenteroides B-1299, a 70% similarity with DSRS from L. mesenteroides B-512 (F, FMCM) and a 45-56% similarity with Streptococcal GTFs.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|