de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris.
Candida antarctica lipase B (CALB) is one of the most widely used and studied enzymes in the world. In order to achieve the high-level expression of CALB in Pichia, we optimized the codons of CALB gene and α-factor by using a de novo design and synthesis strategy. Through comparative analysis of a s...
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ftdoajarticles:oai:doaj.org/article:9d92cdf040f34c269fed8d375fb80b96 2023-05-15T13:52:47+02:00 de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris. Jiang-Ke Yang Li-Ying Liu Jiang-Hong Dai Qin Li 2013-01-01T00:00:00Z https://doi.org/10.1371/journal.pone.0053939 https://doaj.org/article/9d92cdf040f34c269fed8d375fb80b96 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3542265?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0053939 https://doaj.org/article/9d92cdf040f34c269fed8d375fb80b96 PLoS ONE, Vol 8, Iss 1, p e53939 (2013) Medicine R Science Q article 2013 ftdoajarticles https://doi.org/10.1371/journal.pone.0053939 2022-12-30T21:25:14Z Candida antarctica lipase B (CALB) is one of the most widely used and studied enzymes in the world. In order to achieve the high-level expression of CALB in Pichia, we optimized the codons of CALB gene and α-factor by using a de novo design and synthesis strategy. Through comparative analysis of a series of recombinants with different expression components, we found that the methanol-inducible expression recombinant carrying the codon-optimized α-factor and mature CALB gene (pPIC9KαM-CalBM) has the highest lipase production capacity. After fermentation parameters optimization, the lipase activity and protein content of the recombinant pPIC9KαM-CalBM reached 6,100 U/mL and 3.0 g/L, respectively, in a 5-L fermentor. We believe this strategy could be of special interest due to its capacity to improve the expression level of target gene, and the Pichia transformants carrying the codon-optimized gene had great potential for the industrial-scale production of CALB lipase. Article in Journal/Newspaper Antarc* Antarctica Directory of Open Access Journals: DOAJ Articles PLoS ONE 8 1 e53939 |
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Medicine R Science Q |
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Medicine R Science Q Jiang-Ke Yang Li-Ying Liu Jiang-Hong Dai Qin Li de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris. |
topic_facet |
Medicine R Science Q |
description |
Candida antarctica lipase B (CALB) is one of the most widely used and studied enzymes in the world. In order to achieve the high-level expression of CALB in Pichia, we optimized the codons of CALB gene and α-factor by using a de novo design and synthesis strategy. Through comparative analysis of a series of recombinants with different expression components, we found that the methanol-inducible expression recombinant carrying the codon-optimized α-factor and mature CALB gene (pPIC9KαM-CalBM) has the highest lipase production capacity. After fermentation parameters optimization, the lipase activity and protein content of the recombinant pPIC9KαM-CalBM reached 6,100 U/mL and 3.0 g/L, respectively, in a 5-L fermentor. We believe this strategy could be of special interest due to its capacity to improve the expression level of target gene, and the Pichia transformants carrying the codon-optimized gene had great potential for the industrial-scale production of CALB lipase. |
format |
Article in Journal/Newspaper |
author |
Jiang-Ke Yang Li-Ying Liu Jiang-Hong Dai Qin Li |
author_facet |
Jiang-Ke Yang Li-Ying Liu Jiang-Hong Dai Qin Li |
author_sort |
Jiang-Ke Yang |
title |
de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris. |
title_short |
de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris. |
title_full |
de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris. |
title_fullStr |
de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris. |
title_full_unstemmed |
de novo design and synthesis of Candida antarctica lipase B gene and α-factor leads to high-level expression in Pichia pastoris. |
title_sort |
de novo design and synthesis of candida antarctica lipase b gene and α-factor leads to high-level expression in pichia pastoris. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doi.org/10.1371/journal.pone.0053939 https://doaj.org/article/9d92cdf040f34c269fed8d375fb80b96 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
PLoS ONE, Vol 8, Iss 1, p e53939 (2013) |
op_relation |
http://europepmc.org/articles/PMC3542265?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0053939 https://doaj.org/article/9d92cdf040f34c269fed8d375fb80b96 |
op_doi |
https://doi.org/10.1371/journal.pone.0053939 |
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PLoS ONE |
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8 |
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e53939 |
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1766257480922300416 |