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Molecular cloning and characterization of active truncated dextransucrase from Leuconostoc mesenteroides B-1299CB4.
- Source :
-
Bioprocess and biosystems engineering [Bioprocess Biosyst Eng] 2013 Jun; Vol. 36 (6), pp. 857-65. Date of Electronic Publication: 2013 Apr 03. - Publication Year :
- 2013
-
Abstract
- The open reading frame of dsrE563, a dextransucrase gene obtained from a constitutive mutant (CB4-BF563) of Leuconostoc mesenteroides B-1299, consists of 8,511 bp encoding 2,836 amino acid residues. DsrE563 contains two catalytic domains (CD1 and CD2). Two truncated derivative mutants DsrE563ΔCD2ΔGBD (DsrE563-1) and DsrE563ΔCD2ΔVR (DsrE563-2) of DsrE563 were constructed and expressed using the pRSETC vector in Escherichia coli. The derivatives DsrE563-1 (deletion of 1,620 amino acids from the C-terminus) and DsrE563-2 (deletion of 1,258 amino acids from the C-terminus and 349 amino acids from the N-terminus) were expressed as active enzymes. Both enzymes synthesized less-soluble dextran, mainly containing α-1,6 glucosidic linkage. The synthesized less-soluble dextran also had a branched α-1,3 linkage. DsrE563-2 showed 4.5-fold higher dextransucrase activity than that of DsrE563-1 and showed higher acceptor reaction efficiency than that of dextransucrase from L. mesenteroides 512 FMCM when various mono or disaccharides were used as acceptors. Thus, the glucan-binding domain was important for both enzyme expression and dextransucrase activity.
- Subjects :
- Base Sequence
Escherichia coli enzymology
Escherichia coli genetics
Gene Expression
Molecular Sequence Data
Open Reading Frames
Protein Structure, Tertiary
Recombinant Proteins biosynthesis
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins isolation & purification
Amino Acid Sequence
Bacterial Proteins biosynthesis
Bacterial Proteins chemistry
Bacterial Proteins genetics
Bacterial Proteins isolation & purification
Cloning, Molecular
Glucosyltransferases biosynthesis
Glucosyltransferases chemistry
Glucosyltransferases genetics
Glucosyltransferases isolation & purification
Leuconostoc enzymology
Leuconostoc genetics
Sequence Deletion
Subjects
Details
- Language :
- English
- ISSN :
- 1615-7605
- Volume :
- 36
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Bioprocess and biosystems engineering
- Publication Type :
- Academic Journal
- Accession number :
- 23549744
- Full Text :
- https://doi.org/10.1007/s00449-013-0933-3