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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...

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Autores principales: Salleh, Abu Bakar, Baharuddin, Siti Marha, Rahman, Raja Noor Zaliha Raja Abd, Leow, Thean Chor, Basri, Mahiran, Oslan, Siti Nurbaya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7694529/
https://www.ncbi.nlm.nih.gov/pubmed/33171893
http://dx.doi.org/10.3390/microorganisms8111738
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author Salleh, Abu Bakar
Baharuddin, Siti Marha
Rahman, Raja Noor Zaliha Raja Abd
Leow, Thean Chor
Basri, Mahiran
Oslan, Siti Nurbaya
author_facet Salleh, Abu Bakar
Baharuddin, Siti Marha
Rahman, Raja Noor Zaliha Raja Abd
Leow, Thean Chor
Basri, Mahiran
Oslan, Siti Nurbaya
author_sort Salleh, Abu Bakar
collection PubMed
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 P(FLD1). It could also be promising to express other recombinant proteins without inducers.
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spelling pubmed-76945292020-11-28 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 Rahman, Raja Noor Zaliha Raja Abd Leow, Thean Chor Basri, Mahiran Oslan, Siti Nurbaya Microorganisms Article 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 P(FLD1). It could also be promising to express other recombinant proteins without inducers. MDPI 2020-11-06 /pmc/articles/PMC7694529/ /pubmed/33171893 http://dx.doi.org/10.3390/microorganisms8111738 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Salleh, Abu Bakar
Baharuddin, Siti Marha
Rahman, Raja Noor Zaliha Raja Abd
Leow, 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7694529/
https://www.ncbi.nlm.nih.gov/pubmed/33171893
http://dx.doi.org/10.3390/microorganisms8111738
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