Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production
Abstract An Escherichia coli strain expressing γ-glutamyltranspeptidase on its extracellular surface using the Met1 to Arg232 fragment of YiaT of E. coli as an anchor protein was immobilized with alginate for repeated use. Measurement of γ-glutamyltranspeptidase activity of the immobilized cells was...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2023-03-01
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| Series: | AMB Express |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13568-023-01528-9 |
| _version_ | 1797863494253543424 |
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| author | Shintaro Arai Hideyuki Suzuki |
| author_facet | Shintaro Arai Hideyuki Suzuki |
| author_sort | Shintaro Arai |
| collection | DOAJ |
| description | Abstract An Escherichia coli strain expressing γ-glutamyltranspeptidase on its extracellular surface using the Met1 to Arg232 fragment of YiaT of E. coli as an anchor protein was immobilized with alginate for repeated use. Measurement of γ-glutamyltranspeptidase activity of the immobilized cells was performed repeatedly at pH 8.73 and 37 °C for 10 days using γ-glutamyl-p-nitroanilide in the presence of 100 mM CaCl2 and 3% NaCl with and without glycylglycine. Even after the 10th day, the enzyme activity did not decrease from the initial levels. The production of γ-glutamylglutamine from glutamine using the immobilized cells was performed repeatedly at pH 10.5 and 37 °C for 10 days in the presence of 250 mM glutamine, 100 mM CaCl2, and 3% NaCl. Sixty-four % of glutamine was converted to γ-glutamylglutamine in the first cycle. While repeating the production 10 times, the surface of the beads gradually became covered with white precipitate, and the conversion efficiency gradually decreased, but 72% of the initial value still remained even at the 10th measurement. |
| first_indexed | 2024-04-09T22:36:23Z |
| format | Article |
| id | doaj.art-b428faf2272547e6b28ac9e99c0edd43 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-04-09T22:36:23Z |
| publishDate | 2023-03-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-b428faf2272547e6b28ac9e99c0edd432023-03-22T12:27:57ZengSpringerOpenAMB Express2191-08552023-03-011311710.1186/s13568-023-01528-9Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound productionShintaro Arai0Hideyuki Suzuki1Division of Applied Biology, Kyoto Institute of TechnologyDivision of Applied Biology, Kyoto Institute of TechnologyAbstract An Escherichia coli strain expressing γ-glutamyltranspeptidase on its extracellular surface using the Met1 to Arg232 fragment of YiaT of E. coli as an anchor protein was immobilized with alginate for repeated use. Measurement of γ-glutamyltranspeptidase activity of the immobilized cells was performed repeatedly at pH 8.73 and 37 °C for 10 days using γ-glutamyl-p-nitroanilide in the presence of 100 mM CaCl2 and 3% NaCl with and without glycylglycine. Even after the 10th day, the enzyme activity did not decrease from the initial levels. The production of γ-glutamylglutamine from glutamine using the immobilized cells was performed repeatedly at pH 10.5 and 37 °C for 10 days in the presence of 250 mM glutamine, 100 mM CaCl2, and 3% NaCl. Sixty-four % of glutamine was converted to γ-glutamylglutamine in the first cycle. While repeating the production 10 times, the surface of the beads gradually became covered with white precipitate, and the conversion efficiency gradually decreased, but 72% of the initial value still remained even at the 10th measurement.https://doi.org/10.1186/s13568-023-01528-9γ-glutamyl peptideKokumiγ-glutamyltransferaseγ-glutamylglutamineAlginate |
| spellingShingle | Shintaro Arai Hideyuki Suzuki Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production AMB Express γ-glutamyl peptide Kokumi γ-glutamyltransferase γ-glutamylglutamine Alginate |
| title | Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production |
| title_full | Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production |
| title_fullStr | Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production |
| title_full_unstemmed | Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production |
| title_short | Immobilization of E. coli expressing γ-glutamyltranspeptidase on its surface for γ-glutamyl compound production |
| title_sort | immobilization of e coli expressing γ glutamyltranspeptidase on its surface for γ glutamyl compound production |
| topic | γ-glutamyl peptide Kokumi γ-glutamyltransferase γ-glutamylglutamine Alginate |
| url | https://doi.org/10.1186/s13568-023-01528-9 |
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