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Functional characterization of the sucrose isomerase responsible for trehalulose production in plant-associated Pectobacterium species.
- Source :
-
Enzyme and microbial technology [Enzyme Microb Technol] 2014 Feb 05; Vol. 55, pp. 100-6. Date of Electronic Publication: 2013 Oct 24. - Publication Year :
- 2014
-
Abstract
- Fifty-three plant-associated microorganisms were investigated for their ability to convert sucrose to its isomers. These microorganisms included one Dickeya zeae isolate and 7 Enterobacter, 3 Pantoea, and 43 Pectobacterium species. Eleven out of the 53 strains (21%) showed the ability to transform sucrose to isomaltulose and trehalulose. Among those, Pectobacterium carotovorum KKH 3-1 showed the highest bioconversion yield (97.4%) from sucrose to its isomers. In this strain, the addition of up to 14% sucrose in the medium enhanced sucrose isomerase (SIase) production. The SIase activity at 14% sucrose (47.6 U/mg dcw) was about 3.6-fold higher than that of the negative control (13.3 U/mg dcw at 0% sucrose). The gene encoding SIase, which is comprised a 1776 bp open reading frame (ORF) encoding 591 amino acids, was cloned from P. carotovorum KKH 3-1 and expressed in Escherichia coli. The recombinant SIase (PCSI) was shown to have optimum activity at pH 6.0 and 40 °C. The reaction temperature significantly affected the ratio of sucrose isomers produced by PCSI. The amount of trehalulose increased from 47.5% to 79.1% as temperature was lowered from 50 °C to 30 °C, implying that SIase activity can be controlled by reaction temperature.<br /> (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Subjects :
- Amino Acid Sequence
Bacteria enzymology
Bacterial Proteins genetics
Bacterial Proteins metabolism
Base Sequence
Biocatalysis
Cloning, Molecular
Consensus Sequence
Crops, Agricultural microbiology
Escherichia coli
Glucosyltransferases genetics
Glucosyltransferases metabolism
Hydrogen-Ion Concentration
Isomaltose analogs & derivatives
Isomaltose metabolism
Isomerism
Molecular Sequence Data
Pectobacterium carotovorum genetics
Recombinant Fusion Proteins metabolism
Republic of Korea
Sequence Alignment
Sequence Homology, Amino Acid
Species Specificity
Temperature
Bacterial Proteins isolation & purification
Disaccharides biosynthesis
Glucosyltransferases isolation & purification
Pectobacterium carotovorum enzymology
Sucrose metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1879-0909
- Volume :
- 55
- Database :
- MEDLINE
- Journal :
- Enzyme and microbial technology
- Publication Type :
- Academic Journal
- Accession number :
- 24411451
- Full Text :
- https://doi.org/10.1016/j.enzmictec.2013.10.005