A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB
Screening for a new yeast as an alternative host is expected to solve the limitations in the present yeast expression system. A yeast sample which was isolated from the traditional food starter ‘ragi’ from Malaysia was identified to contain Meyerozyma guilliermondii strain SMB. This yeast-like fungu...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Multidisciplinary Digital Publishing Institute
2020
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| Online Access: | http://psasir.upm.edu.my/id/eprint/87604/1/ABSTRACT.pdf |
| _version_ | 1796981864998633472 |
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| author | Salleh, Abu Bakar Baharuddin, Siti Marha Raja Abd Rahman, Raja Noor Zaliha Leow, Adam Thean Chor Basri, Mahiran Oslan, Siti Nurbaya |
| author_facet | Salleh, Abu Bakar Baharuddin, Siti Marha Raja Abd Rahman, Raja Noor Zaliha Leow, Adam Thean Chor Basri, Mahiran Oslan, Siti Nurbaya |
| author_sort | Salleh, Abu Bakar |
| collection | UPM |
| description | Screening for a new yeast as an alternative host is expected to solve the limitations in the present yeast expression system. A yeast sample which was isolated from the traditional food starter ‘ragi’ from Malaysia was identified to contain Meyerozyma guilliermondii strain SMB. This yeast-like fungus strain SMB was characterized to assess its suitability as an expression host. Lipase activity was absent in this host (when assayed at 30 °C and 70 °C) and Hygromycin B (50 μg/mL) was found to be its best selection marker. Then, the hyg gene (Hygromycin B) was used to replace the sh ble gene (Zeocin) expression cassette in a Komagataella phaffii expression vector (designated as pFLDhα). A gene encoding the mature thermostable lipase from Bacillus sp. L2 was cloned into pFLDhα, followed by transformation into strain SMB. The optimal expression of L2 lipase was achieved using YPTM (Yeast Extract-Peptone-Tryptic-Methanol) medium after 48 h with 0.5% (v/v) methanol induction, which was 3 times faster than another K. phaffii expression system. In conclusion, a new host-vector system was established as a platform to express L2 lipase under the regulation of PFLD1. It could also be promising to express other recombinant proteins without inducers |
| first_indexed | 2024-03-06T10:43:53Z |
| format | Article |
| id | upm.eprints-87604 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T10:43:53Z |
| publishDate | 2020 |
| publisher | Multidisciplinary Digital Publishing Institute |
| record_format | dspace |
| spelling | upm.eprints-876042022-07-06T08:20:33Z http://psasir.upm.edu.my/id/eprint/87604/ A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB Salleh, Abu Bakar Baharuddin, Siti Marha Raja Abd Rahman, Raja Noor Zaliha Leow, Adam Thean Chor Basri, Mahiran Oslan, Siti Nurbaya Screening for a new yeast as an alternative host is expected to solve the limitations in the present yeast expression system. A yeast sample which was isolated from the traditional food starter ‘ragi’ from Malaysia was identified to contain Meyerozyma guilliermondii strain SMB. This yeast-like fungus strain SMB was characterized to assess its suitability as an expression host. Lipase activity was absent in this host (when assayed at 30 °C and 70 °C) and Hygromycin B (50 μg/mL) was found to be its best selection marker. Then, the hyg gene (Hygromycin B) was used to replace the sh ble gene (Zeocin) expression cassette in a Komagataella phaffii expression vector (designated as pFLDhα). A gene encoding the mature thermostable lipase from Bacillus sp. L2 was cloned into pFLDhα, followed by transformation into strain SMB. The optimal expression of L2 lipase was achieved using YPTM (Yeast Extract-Peptone-Tryptic-Methanol) medium after 48 h with 0.5% (v/v) methanol induction, which was 3 times faster than another K. phaffii expression system. In conclusion, a new host-vector system was established as a platform to express L2 lipase under the regulation of PFLD1. It could also be promising to express other recombinant proteins without inducers Multidisciplinary Digital Publishing Institute 2020 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/87604/1/ABSTRACT.pdf Salleh, Abu Bakar and Baharuddin, Siti Marha and Raja Abd Rahman, Raja Noor Zaliha and Leow, Adam Thean Chor and Basri, Mahiran and Oslan, Siti Nurbaya (2020) A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB. Microorganisms, 8 (11). art. no. 1738. pp. 1-11. ISSN 2076-2607 https://www.mdpi.com/2076-2607/8/11/1738 10.3390/microorganisms8111738 |
| spellingShingle | Salleh, Abu Bakar Baharuddin, Siti Marha Raja Abd Rahman, Raja Noor Zaliha Leow, Adam Thean Chor Basri, Mahiran Oslan, Siti Nurbaya A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB |
| title | A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB |
| title_full | A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB |
| title_fullStr | A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB |
| title_full_unstemmed | A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB |
| title_short | A host-vector system for the expression of a thermostable bacterial lipase in a locally isolated Meyerozyma guilliermondii SMB |
| title_sort | host vector system for the expression of a thermostable bacterial lipase in a locally isolated meyerozyma guilliermondii smb |
| url | http://psasir.upm.edu.my/id/eprint/87604/1/ABSTRACT.pdf |
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