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Expression and activity analysis of sucrose:sucrose 1-fructosyltransferase from onion
- Source :
- New biotechnology. 27(4)
- Publication Year :
- 2009
-
Abstract
- This study was designed to express the onion fructosyltransferase by Escherichia coli DH5alpha, and obtain the optimal conditions of FST-1 activity. Thereby, fructosyltransferase gene was obtained by RT-PCR from onion in this experiment, and named FST-1. The expressed proteins were analyzed by SDS-PAGE. FST-1 activity was identified by the high performance liquid chromatography (HPLC). The optimal conditions of FST-1 were analyzed by the dinonylnaphthalene sulfonic acid (DNS) and orthogonal test. Results revealed that FST-1 was identified to 98% similarity with fructosyltransferase mRNA of onion (accession number: AJ006066). FST-1 was successfully expressed in E. coli DH5alpha. HPLC results indicated that the expressed protein from FST-1 had a good transferring activity for fructose. The optimal conditions of FST-1 in catalyzing reaction were the pH 5.0, 45 degrees C and 60% sucrose substrate. The results in this experiment would lay the foundation for the large-scale of kestose by bio-catalysis method.
- Subjects :
- Sucrose
Time Factors
Bioengineering
medicine.disease_cause
High-performance liquid chromatography
Gene Expression Regulation, Enzymologic
Substrate Specificity
chemistry.chemical_compound
Gene Expression Regulation, Plant
Gene expression
Onions
medicine
Molecular Biology
Escherichia coli
Chromatography, High Pressure Liquid
biology
Accession number (library science)
Reverse Transcriptase Polymerase Chain Reaction
Temperature
Substrate (chemistry)
Fructose
General Medicine
Hydrogen-Ion Concentration
Reference Standards
Enzyme assay
chemistry
Biochemistry
Hexosyltransferases
biology.protein
human activities
Biotechnology
Subjects
Details
- ISSN :
- 18764347
- Volume :
- 27
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- New biotechnology
- Accession number :
- edsair.doi.dedup.....6775a8d41cd56fa8685346c73e7053b2