Cloning and expression of sucrose phosphorylase gene from Bifidobacterium longum in E. coli and characterization of the recombinant enzyme

Mijin Kim, Taeyeon Kwon, Hyong Joo Lee, Kyoung Heon Kim, Dae Kyun Chung, Geun Eog Ji, Eui Seok Byeon, Jong Hoon Lee

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)

Abstract

A DNA fragment, which complemented the growth of E. coli both on M9 medium containing raffinose and on LB medium containing ampicillin, IPTG and 5-bromo-4-chloro-3-indoxyl-α-D-galactoside, was isolated from the genomic library of Bifidobacterium longum SJ32, which had been digested with EcoRI. In the cloned DNA fragment, a gene encoding a sucrose phosphorylase (splP) and a partially cloned putative sucrose regulator gene (splR) were identified using the deletion analysis and sequence analysis. A 56 kDa protein was synthesized in E. coli and partially purified by DEAE-ion exchange chromatography. The partially purified enzyme did not react with melibiose, melezitoze and raffinose but did with sucrose. It had transglucosylation activity in addition to hydrolytic activity.

Original languageEnglish
Pages (from-to)1211-1217
Number of pages7
JournalBiotechnology letters
Volume25
Issue number15
DOIs
Publication statusPublished - 2003 Aug
Externally publishedYes

Keywords

  • Bifidobacterium longum
  • SplP
  • SplR
  • Sucrose phosphorylase
  • Transglycosylation
  • α-linked oligosaccharide

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'Cloning and expression of sucrose phosphorylase gene from Bifidobacterium longum in E. coli and characterization of the recombinant enzyme'. Together they form a unique fingerprint.

Cite this