Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst.
Yeast Surface Display (YSD) is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB) is one of the most studied enzymes in the context of industrial biotechnology. This study...
Published in: | PLOS ONE |
---|---|
Main Authors: | , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Public Library of Science (PLoS)
2015
|
Subjects: | |
Online Access: | https://doi.org/10.1371/journal.pone.0141454 https://doaj.org/article/23d2736ded32498e99b9ff6ddafe08b1 |
id |
ftdoajarticles:oai:doaj.org/article:23d2736ded32498e99b9ff6ddafe08b1 |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:23d2736ded32498e99b9ff6ddafe08b1 2023-05-15T13:59:22+02:00 Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst. Marcelo Victor Holanda Moura Giulia Pontes da Silva Antônio Carlos de Oliveira Machado Fernando Araripe Gonçalves Torres Denise Maria Guimarães Freire Rodrigo Volcan Almeida 2015-01-01T00:00:00Z https://doi.org/10.1371/journal.pone.0141454 https://doaj.org/article/23d2736ded32498e99b9ff6ddafe08b1 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4624902?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0141454 https://doaj.org/article/23d2736ded32498e99b9ff6ddafe08b1 PLoS ONE, Vol 10, Iss 10, p e0141454 (2015) Medicine R Science Q article 2015 ftdoajarticles https://doi.org/10.1371/journal.pone.0141454 2022-12-31T10:28:11Z Yeast Surface Display (YSD) is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB) is one of the most studied enzymes in the context of industrial biotechnology. This study aimed to assess the biochemical features of this important biocatalyst when immobilized on the cell surface of the methylotrophic yeast Pichia pastoris using the YSD approach. For that purpose, two anchors were tested. The first (Flo9) was identified after a prospection of the P. pastoris genome being related to the family of flocculins similar to Flo1 but significantly smaller. The second is the Protein with Internal Repeats (Pir1) from P. pastoris. An immunolocalization assay showed that both anchor proteins were able to display the reporter protein EGFP in the yeast outer cell wall. LipB was expressed in P. pastoris fused either to Flo9 (FLOLIPB) or Pir1 (PIRLIPB). Both constructions showed hydrolytic activity towards tributyrin (>100 U/mgdcw and >80 U/mgdcw, respectively), optimal hydrolytic activity around 45°C and pH 7.0, higher thermostability at 45°C and stability in organic solvents when compared to a free lipase. Article in Journal/Newspaper Antarc* Antarctica Directory of Open Access Journals: DOAJ Articles PLOS ONE 10 10 e0141454 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Medicine R Science Q |
spellingShingle |
Medicine R Science Q Marcelo Victor Holanda Moura Giulia Pontes da Silva Antônio Carlos de Oliveira Machado Fernando Araripe Gonçalves Torres Denise Maria Guimarães Freire Rodrigo Volcan Almeida Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst. |
topic_facet |
Medicine R Science Q |
description |
Yeast Surface Display (YSD) is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB) is one of the most studied enzymes in the context of industrial biotechnology. This study aimed to assess the biochemical features of this important biocatalyst when immobilized on the cell surface of the methylotrophic yeast Pichia pastoris using the YSD approach. For that purpose, two anchors were tested. The first (Flo9) was identified after a prospection of the P. pastoris genome being related to the family of flocculins similar to Flo1 but significantly smaller. The second is the Protein with Internal Repeats (Pir1) from P. pastoris. An immunolocalization assay showed that both anchor proteins were able to display the reporter protein EGFP in the yeast outer cell wall. LipB was expressed in P. pastoris fused either to Flo9 (FLOLIPB) or Pir1 (PIRLIPB). Both constructions showed hydrolytic activity towards tributyrin (>100 U/mgdcw and >80 U/mgdcw, respectively), optimal hydrolytic activity around 45°C and pH 7.0, higher thermostability at 45°C and stability in organic solvents when compared to a free lipase. |
format |
Article in Journal/Newspaper |
author |
Marcelo Victor Holanda Moura Giulia Pontes da Silva Antônio Carlos de Oliveira Machado Fernando Araripe Gonçalves Torres Denise Maria Guimarães Freire Rodrigo Volcan Almeida |
author_facet |
Marcelo Victor Holanda Moura Giulia Pontes da Silva Antônio Carlos de Oliveira Machado Fernando Araripe Gonçalves Torres Denise Maria Guimarães Freire Rodrigo Volcan Almeida |
author_sort |
Marcelo Victor Holanda Moura |
title |
Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst. |
title_short |
Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst. |
title_full |
Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst. |
title_fullStr |
Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst. |
title_full_unstemmed |
Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst. |
title_sort |
displaying lipase b from candida antarctica in pichia pastoris using the yeast surface display approach: prospection of a new anchor and characterization of the whole cell biocatalyst. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2015 |
url |
https://doi.org/10.1371/journal.pone.0141454 https://doaj.org/article/23d2736ded32498e99b9ff6ddafe08b1 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
PLoS ONE, Vol 10, Iss 10, p e0141454 (2015) |
op_relation |
http://europepmc.org/articles/PMC4624902?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0141454 https://doaj.org/article/23d2736ded32498e99b9ff6ddafe08b1 |
op_doi |
https://doi.org/10.1371/journal.pone.0141454 |
container_title |
PLOS ONE |
container_volume |
10 |
container_issue |
10 |
container_start_page |
e0141454 |
_version_ |
1766267909883035648 |