Immobilization of transferase enzymes for the production of isooligosaccharide panose | |
| Author | Vorapat Sanguanchaipaiwong |
| Call Number | AIT Thesis no. BP-00-08 |
| Subject(s) | Oligosaccharides Glycosyltransferases Glycosidases |
| Note | A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science, School of Environment, Resources and Development |
| Publisher | Asian Institute of Technology |
| Series Statement | Thesis ; no. BP-00-08 |
| Abstract | The main aim of this study was to immobilize one fraction of transferase enzymes from the fermentation broth of Aspergillus foetidus for the production of isooligosaccharide panose. Optimization of conditions for the production of isomaltooligosaccharide panose using this fraction compared to free enzyme and immobilized freeze-dried enzyme obtained from ammonium sulphate precipitation were done. The transferase enzymes were produced by Aspergillus foetidus NRRL 337 fermented for 7 days. After the fermentation broth was separated by ammonium sulphate precipitation (45% - 75% saturation), crude enzyme was obtained. The fractions having transferase enzyme activity were obtained from freeeze-dried enyme obtained from ammonium sulphate precipitation by DEAE-cellulose chromatography. Freeeze-dried enyme obtained from ammonium sulphate precipitation and one fraction of transferase enzyme (fraction A) were ir'nmobilized into ·calcium alginate beads. The immobilization efficiencies of immobilized freeeze-dried enyme obtained from ammonium sulphate precipitation and immobilized fraction A transferase enzyme were 37% and 19%, respectively. PEG was not found to give fractionation of the transferase enzymes_in this study. Optimization of conditions (i.e. pH, temperature, and stability) for isomaltooligosaccharide was done. Optimum pH and temperature of immobilized fraction A transferase enzyme were 4.5 and 50 °C. Maltose concentration of 30% gave the best panose production. The immobilized fraction A transferase enzyme showed improved kinetics, pH and temperature stability, etc. compared to free enzyme. The immobilized fraction A transferase enzyme was tested for 4 cycles and it was found that reuse of the enzyme was possible. The Michaelis-Menten constant of immobilized fraction A transferase enzyme was less than those of immobilized freeeze-dried enyme obtained from ammonium sulphate precipitation and free transferase enzyme fraction A showing that the enzyme in this form had greater affinity for the substrate. |
| Year | 2000 |
| Corresponding Series Added Entry | Asian Institute of Technology. Thesis ; no. BP-00-08 |
| Type | Thesis |
| School | School of Environment, Resources, and Development (SERD) |
| Department | Department of Food, Agriculture and Natural Resources (Former title: Department of Food Agriculture, and BioResources (DFAB)) |
| Academic Program/FoS | Bioprocess Technology (BP) |
| Chairperson(s) | Rakshit, Sudip K.; |
| Examination Committee(s) | Stevens, Willem F.;Suwalee Chandrkrachang;Parkon Nuchnoi; |
| Scholarship Donor(s) | Partial Scholarships; |
| Degree | Thesis (M.Sc.) - Asian Institute of Technology, 2000 |