Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium
Lactose is a disaccharide found in most dairy products. Parallel to this a large percentage of the global population suffers from hypolactasia and is intolerant to the substance. The dairy industry is constantly developing new products that are free from lactose, to satisfy these individuals. Even t...
| Main Author: | |
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| Format: | M2 |
| Language: | Inglés |
| Published: |
SLU/Dept. of Microbiology
2015
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| Subjects: |
| _version_ | 1855571377747132416 |
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| author | Borsch Reniers, Henrik |
| author_browse | Borsch Reniers, Henrik |
| author_facet | Borsch Reniers, Henrik |
| author_sort | Borsch Reniers, Henrik |
| collection | Epsilon Archive for Student Projects |
| description | Lactose is a disaccharide found in most dairy products. Parallel to this a large percentage of the global population suffers from hypolactasia and is intolerant to the substance. The dairy industry is constantly developing new products that are free from lactose, to satisfy these individuals. Even though producers have come a long way in developing techniques that are efficient and cheap there is still improvements to be made. β-D-galactosidase have been proven to effectively remove lactase from dairy products, still further enhancement is presumed possible.
In this study the aim was to further explore and develop the properties of known β-D-galactosidase enzyme found in Arthrobacter psychrolactophilus. With the use of polymerase chain reaction and transformation techniques, the β-D-galactosidase coding bglA gene from Arthrobacter psychrolactophilus B7 was transformed into Escherichia coli BL 21 competent cells. Furthermore over expression of the rBglAp and spectrophotometric analysis was conducted to detect any successful transformations.
The transformation showed no indication of catalysis of lactose into its monomeric forms. Further verification of the techniques would have increased the success rate of the experiment. Detection methods like subcloning of the plasmid vector and selective media should be performed if the experiment is repeated. |
| format | M2 |
| id | RepoSLU8592 |
| institution | Swedish University of Agricultural Sciences |
| language | Inglés |
| publishDate | 2015 |
| publishDateSort | 2015 |
| publisher | SLU/Dept. of Microbiology |
| publisherStr | SLU/Dept. of Microbiology |
| record_format | eprints |
| spelling | RepoSLU85922015-10-27T12:09:46Z Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium Borsch Reniers, Henrik Lactase Arthrobacter sp Beta-galactosidase Lactose is a disaccharide found in most dairy products. Parallel to this a large percentage of the global population suffers from hypolactasia and is intolerant to the substance. The dairy industry is constantly developing new products that are free from lactose, to satisfy these individuals. Even though producers have come a long way in developing techniques that are efficient and cheap there is still improvements to be made. β-D-galactosidase have been proven to effectively remove lactase from dairy products, still further enhancement is presumed possible. In this study the aim was to further explore and develop the properties of known β-D-galactosidase enzyme found in Arthrobacter psychrolactophilus. With the use of polymerase chain reaction and transformation techniques, the β-D-galactosidase coding bglA gene from Arthrobacter psychrolactophilus B7 was transformed into Escherichia coli BL 21 competent cells. Furthermore over expression of the rBglAp and spectrophotometric analysis was conducted to detect any successful transformations. The transformation showed no indication of catalysis of lactose into its monomeric forms. Further verification of the techniques would have increased the success rate of the experiment. Detection methods like subcloning of the plasmid vector and selective media should be performed if the experiment is repeated. SLU/Dept. of Microbiology 2015 M2 eng https://stud.epsilon.slu.se/8592/ |
| spellingShingle | Lactase Arthrobacter sp Beta-galactosidase Borsch Reniers, Henrik Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium |
| title | Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium |
| title_full | Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium |
| title_fullStr | Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium |
| title_full_unstemmed | Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium |
| title_short | Cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium |
| title_sort | cloning and in vitro evolution of a beta-galactosidase from a psychrophilic bacterium |
| topic | Lactase Arthrobacter sp Beta-galactosidase |