HOMEPRODUCTSCOMPANYCONTACTFAQResearchDictionaryPharmaSign Up FREE or Login

Molecular analysis of a Clostridium butyricum NCIMB 7423 gene encoding 4-alpha-glucanotransferase and characterization of the recombinant enzyme produced in Escherichia coli.

Abstract
An Escherichia coli clone was detected in a Clostridium butyricum NCIMB 7423 plasmid library capable of degrading soluble amylose. Deletion subcloning of its recombinant plasmid indicated that the gene(s) responsible for amylose degradation was localized on a 1.8 kb NspHI-Scal fragment. This region was sequenced in its entirety and shown to encompass a large ORF capable of encoding a protein with a calculated molecular mass of 57,184 Da. Although the deduced amino acid sequence showed only weak similarity with known amylases, significant sequences identity was apparent with the 4-alpha-glucano-transferase enzymes of Streptococcus pneumoniae (46.9%), potato (42.9%) and E. coli (16.2%). The clostridial gene (designated maIQ) was followed by a second ORF which, through its homology to the equivalent enzymes of E. coli and S. pneumoniae, was deduced to encode maltodextrin phosphorylase (MaIP). The translation stop codon of MaIQ overlapped the translation start codon of the putative maIP gene, suggesting that the two genes may be both transcriptionally and translationally coupled. 4-alpha-Glucanotransferase catalyses a disproportionation reaction in which single or multiple glucose units from oligosaccharides are transferred to the 4-hydroxyl group of acceptor sugars. Characterization of the recombinant C. butyricum enzyme demonstrated that glucose, maltose and maltotriose could act as acceptor, whereas of the three only maltotriose could act as donor. The enzyme therefore shares properties with the E. coli MaIQ protein, but differs significantly from the glucanotransferase of Thermotoga maritima, which is unable to use maltotriose as donor or glucose as acceptor. Physiologically, the concerted action of 4-alpha-glucanotransferase and maltodextrin phosphorylase provides C. butyricum with a mechanism of utilizing amylose/maltodextrins with little drain on cellular ATP reserves.
AuthorsSayed K Goda, Omima Eissa, Muhammad Akhtar, Nigel P Minton
JournalMicrobiology (Reading, England) (Microbiology (Reading)) Vol. 143 ( Pt 10) Pg. 3287-3294 (Oct 1997) ISSN: 1350-0872 [Print] England
PMID9353929 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • DNA, Bacterial
  • Glycogen Debranching Enzyme System
  • Recombinant Proteins
  • Amylose
  • Glucosyltransferases
  • maltodextrin phosphorylase
  • 4 alpha-glucanotransferase
Topics
  • Amino Acid Sequence
  • Amylose (metabolism)
  • Base Sequence
  • Cloning, Molecular
  • Clostridium (enzymology, genetics)
  • DNA, Bacterial (genetics)
  • Escherichia coli (enzymology, genetics)
  • Genes, Bacterial
  • Glucosyltransferases (genetics)
  • Glycogen Debranching Enzyme System (biosynthesis, genetics, metabolism)
  • Molecular Sequence Data
  • Molecular Weight
  • Open Reading Frames
  • Recombinant Proteins (biosynthesis, genetics)
  • Sequence Homology, Amino Acid

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.
Realize the full power of the drug-disease research graph!


Choose Username:
Email:
Password:
Verify Password:
Enter Code Shown: