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High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production

BACKGROUND: Production of biodiesel from non-edible oils is receiving increasing attention. Tung oil, called “China wood oil” is one kind of promising non-edible biodiesel oil in China. To our knowledge, tung oil has not been used to produce biodiesel by enzymatic method. The enzymatic production of...

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Autores principales: Yu, Xiao-Wei, Sha, Chong, Guo, Yong-Liang, Xiao, Rong, Xu, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3674748/
https://www.ncbi.nlm.nih.gov/pubmed/23432946
http://dx.doi.org/10.1186/1754-6834-6-29
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author Yu, Xiao-Wei
Sha, Chong
Guo, Yong-Liang
Xiao, Rong
Xu, Yan
author_facet Yu, Xiao-Wei
Sha, Chong
Guo, Yong-Liang
Xiao, Rong
Xu, Yan
author_sort Yu, Xiao-Wei
collection PubMed
description BACKGROUND: Production of biodiesel from non-edible oils is receiving increasing attention. Tung oil, called “China wood oil” is one kind of promising non-edible biodiesel oil in China. To our knowledge, tung oil has not been used to produce biodiesel by enzymatic method. The enzymatic production of biodiesel has been investigated extensively by using Rhizopus oryzae lipase as catalyst. However, the high cost of R. oryzae lipase remains a barrier for its industrial applications. Through different heterologous expression strategies and fermentation techniques, the highest expression level of the lipase from R. oryzae reached 1334 U/mL in Pichia pastoris, which is still not optimistic for industry applications. RESULTS: The prosequence of lipases from Rhizopus sp. is very important for the folding and secretion of an active lipase. A chimeric lipase from R. oryzae was constructed by replacing the prosequence with that from the R. chinensis lipase and expressed in P. pastoris. The maximum activity of the chimera reached 4050 U/mL, which was 11 fold higher than that of the parent. The properties of the chimera were studied. The immobilized chimera was used successfully for biodiesel production from tung oil, which achieved higher FAME yield compared with the free chimeric lipase, non-chimeric lipase and mature lipase. By response surface methodology, three variables, water content, methanol to tung oil molar ratio and enzyme dosage were proved to be crucial parameters for biosynthesis of FAME and the FAME yield reached 91.9±2.5% at the optimized conditions by adding 5.66 wt.% of the initial water based on oil weight, 3.88 of methanol to tung oil molar ratio and 13.24 wt.% of enzyme concentration based on oil weight at 40°C. CONCLUSIONS: This is the first report on improving the expression level of the lipase from R. oryzae by replacing prosequences. The immobilized chimera was used successfully for biodiesel production from tung oil. Using tung oil as non-edible raw material and a chimeric lipase from R. oryzae as an economic catalyst make this study a promising one for biodiesel applications.
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spelling pubmed-36747482013-06-10 High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production Yu, Xiao-Wei Sha, Chong Guo, Yong-Liang Xiao, Rong Xu, Yan Biotechnol Biofuels Research BACKGROUND: Production of biodiesel from non-edible oils is receiving increasing attention. Tung oil, called “China wood oil” is one kind of promising non-edible biodiesel oil in China. To our knowledge, tung oil has not been used to produce biodiesel by enzymatic method. The enzymatic production of biodiesel has been investigated extensively by using Rhizopus oryzae lipase as catalyst. However, the high cost of R. oryzae lipase remains a barrier for its industrial applications. Through different heterologous expression strategies and fermentation techniques, the highest expression level of the lipase from R. oryzae reached 1334 U/mL in Pichia pastoris, which is still not optimistic for industry applications. RESULTS: The prosequence of lipases from Rhizopus sp. is very important for the folding and secretion of an active lipase. A chimeric lipase from R. oryzae was constructed by replacing the prosequence with that from the R. chinensis lipase and expressed in P. pastoris. The maximum activity of the chimera reached 4050 U/mL, which was 11 fold higher than that of the parent. The properties of the chimera were studied. The immobilized chimera was used successfully for biodiesel production from tung oil, which achieved higher FAME yield compared with the free chimeric lipase, non-chimeric lipase and mature lipase. By response surface methodology, three variables, water content, methanol to tung oil molar ratio and enzyme dosage were proved to be crucial parameters for biosynthesis of FAME and the FAME yield reached 91.9±2.5% at the optimized conditions by adding 5.66 wt.% of the initial water based on oil weight, 3.88 of methanol to tung oil molar ratio and 13.24 wt.% of enzyme concentration based on oil weight at 40°C. CONCLUSIONS: This is the first report on improving the expression level of the lipase from R. oryzae by replacing prosequences. The immobilized chimera was used successfully for biodiesel production from tung oil. Using tung oil as non-edible raw material and a chimeric lipase from R. oryzae as an economic catalyst make this study a promising one for biodiesel applications. BioMed Central 2013-02-21 /pmc/articles/PMC3674748/ /pubmed/23432946 http://dx.doi.org/10.1186/1754-6834-6-29 Text en Copyright © 2013 Yu et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Yu, Xiao-Wei
Sha, Chong
Guo, Yong-Liang
Xiao, Rong
Xu, Yan
High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production
title High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production
title_full High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production
title_fullStr High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production
title_full_unstemmed High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production
title_short High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production
title_sort high-level expression and characterization of a chimeric lipase from rhizopus oryzae for biodiesel production
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3674748/
https://www.ncbi.nlm.nih.gov/pubmed/23432946
http://dx.doi.org/10.1186/1754-6834-6-29
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