Designing of a novel dextransucrase efficient in acceptor reactions
| Autor: | Mahmut Parlak, Duran Ustek, Aziz Tanriseven |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Recombinant Fusion Proteins
Protein Engineering medicine.disease_cause Biochemistry Analytical Chemistry Dextransucrase Calcium Chloride chemistry.chemical_compound Bacterial Proteins Catalytic Domain Escherichia coli medicine Leuconostoc Glutathione Transferase chemistry.chemical_classification biology Chemistry Organic Chemistry Active site Dextrans General Medicine Hydrogen-Ion Concentration Enzymes Immobilized biology.organism_classification Enzyme assay Kinetics Dextran Enzyme Glucosyltransferases Leuconostoc mesenteroides Biocatalysis biology.protein |
| Zdroj: | Europe PubMed Central |
| ISSN: | 0008-6215 |
| DOI: | 10.1016/j.carres.2014.01.003 |
| Popis: | Dextransucrase is produced by Leuconostoc, Streptococcus and Lactobacillus Species. The enzyme synthesizes dextran and acceptor products some of which act as prebiotics that are increasingly used in such industries as food, medicine, and cosmetics. B-512F Leuconostoc mesenteroides dextransucrase (DSR-S) is the preferred enzyme in commercial production of dextran and prebiotics. In the present work, a novel dextransucrase which is efficient in prebiotics production was designed. The enzyme was produced at optimal conditions in Escherichia coli by truncation and fusion to glutathione S-transferase (GST) in the gene from Leuconostoc mesenteroides B-512 FMC. The novel enzyme (MW: 119 kDa) was active and carried out dextran biosynthesis and acceptor reactions effectively. The novel dextransucrase (fTDSR-S) was produced by truncating signal, variable, and the glucan-binding regions in the gene and fusion of gst gene at the 5' end. fTDSR-S was characterized in detail and compared to the DSR-S. Truncation and fusion resulted in an increase in fTDSR-S biosynthesis in E. coli BL21 (DE3) by 35 fold. fTDSR-S leads to production of dextran as well as increased acceptor reactions. Due to GST fusion, it was possible to immobilize fTDSR-S covalently onto Eupergit C successfully. It was also found that the size of the active site of dextransucrase is 49 amino acids shorter than that reported previously in the literature. |
| Databáze: | OpenAIRE |
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