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Universidade Federal o Rio Grande Do Sul - UFRGS Instituto de Ciência e Tecnologia de Alimentos - ICTA Programa e Pós-Graduação em Ciência e Tecnologia de Alimentos
| Content Provider | Semantic Scholar |
|---|---|
| Author | Wrzesinski, Andiara |
| Copyright Year | 2013 |
| Abstract | Cyclodextrin glucosyltransferase (CGTase) is an industrially important enzyme, capable to convert starch into cyclodextrins (CDs). The CDs are able to form inclusion complexes with a wide range of organic and inorganic compounds, which can change their chemical and physical properties, that gives them extensive applicability in the food, pharmaceutical, chemical, cosmetics and agricultural. Currently, many CGTases have been isolated and characterized from various microorganisms, particularly Bacillus, Paenibacillus, Pseudomonas, Klebsiella, Xanthomonas, Thermococcus, Vibrio, Geobacillus and Thermoanaerobacterium. This work demonstrates the first report involving cloning and expression of heterologous CGTase from Stenotrophomonas maltophilia, microorganism isolated from Brazilian soil. The gene encoding the S. maltophilia CGTase, was successfully amplified by PCR, cloned into the vector pET-23a (+) and expressed in Escherichia coli BL21(DE3). Recombinant cells required about 4 h of cultivation in Luria Bertani (LB) after addition of 0.1 mM IPTG for high expression of the target protein. However, the CGTase was expressed recombinant form of inclusion bodies remaining in the insoluble fraction, being necessary use protocols for solubilization, including different concentrations of urea, but the precipitation was not effective. Although it was observed a high expression of the protein about 60 kDa on SDS-PAGE 12%, corresponding to the expected size of the protein, but the active form of the enzyme was not obtained. Bioinformatic analysis was performed and observed a Putative Major Facilitator Transmembrane Protein (PMFTP) harboring the cgt gene. This protein can be used in further studies to develop a new and more effective system for expression of the CGTase, which may facilitate its extracellular expression. Thus, more studies are needed to develop a system of co-expression of rCGTase::PMFTP in E. coli and acquire more information about this protein of Stenotrophomonas maltophilia. |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://lume.ufrgs.br/bitstream/handle/10183/77643/000886031.pdf;jsessionid=1A2876BEE08FB13FD22D0F3D2F66E6FA?sequence=1 |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
