Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design
Performing transesterifications in aqueous media is becoming a priority challenge in lipid biotechnology in order to develop more eco-friendly and efficient biocatalytic processes in systems containing both polar and apolar substrates. In this context, our group has explored for several years the hi...
Published in: | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics |
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ftinraparis:oai:prodinra.inra.fr:333999 2023-05-15T14:03:09+02:00 Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design Jan, Anne Hélène Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric Subileau, Maeva 2016 application/pdf http://prodinra.inra.fr/ft/9CDAC24F-45A6-47B3-9C56-1003E9BC23C7 http://prodinra.inra.fr/record/333999 https://doi.org/10.1016/j.bbapap.2015.11.006 eng eng http://creativecommons.org/licenses/by-nd-nc/1.0/ CC-BY-ND-NC Biochimica et Biophysica Acta - Proteins and Proteomics 2 (1864), 187–194. (2016) Ingénierie des aliments Food engineering lipases/acyltransferases;CpLIP2;CAL-A;transesterification;rational design;biocatalysis transesterification acétyltransférase activité enzymatique lipogénique biocatalyse candida parapsilosis lipase ARTICLE 2016 ftinraparis https://doi.org/10.1016/j.bbapap.2015.11.006 2016-11-22T23:27:55Z Performing transesterifications in aqueous media is becoming a priority challenge in lipid biotechnology in order to develop more eco-friendly and efficient biocatalytic processes in systems containing both polar and apolar substrates. In this context, our group has explored for several years the high potential of the lipase/acyltransferase CpLIP2 from Candida parapsilosis and of several of its homologs, that catalyze efficiently acyltransfer reactions in lipid/water media with high water activity (aw > 0.9). The discovery of a new member of this group, CduLAc from C. dubliniensis, with a higher acyltransferase activity than CpLIP2, has provided a new insight on structure-function relationships in this group. Indeed, the comparison of sequences and 3D models, especially of CpLIP2 and CduLAc, with those of the phylogenetically related lipase A from Ps. Antarctica (CAL-A), allowed elucidating a key structural determinant of the acyltransferase activity: serine S369 in CpLIP2 and its equivalents E370 in CAL-A and A366 in CduLAc. Mutants obtained by rational design at this key position showed significant changes in acyltransfer activity. Whereas mutation S369E resulted in an increase in the hydrolytic activity of CpLIP2, S369A increased alcoholysis. More strikingly, the single E370A mutation in CAL-A drastically increased the acyltransferase activity of this enzyme, giving it the character of a lipase/acyltransferase. Indeed, this single mutation lowered the methanol concentration for which the initial rates of alcoholysis and hydrolysis are equal from 2 M in CAL-A down to 0.3 M in its mutant, while the exceptional stability of the parental enzyme toward alcohol and temperature was conserved. Article in Journal/Newspaper Antarc* Antarctica Institut National de la Recherche Agronomique: ProdINRA Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics 1864 2 187 194 |
institution |
Open Polar |
collection |
Institut National de la Recherche Agronomique: ProdINRA |
op_collection_id |
ftinraparis |
language |
English |
topic |
Ingénierie des aliments Food engineering lipases/acyltransferases;CpLIP2;CAL-A;transesterification;rational design;biocatalysis transesterification acétyltransférase activité enzymatique lipogénique biocatalyse candida parapsilosis lipase |
spellingShingle |
Ingénierie des aliments Food engineering lipases/acyltransferases;CpLIP2;CAL-A;transesterification;rational design;biocatalysis transesterification acétyltransférase activité enzymatique lipogénique biocatalyse candida parapsilosis lipase Jan, Anne Hélène Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design |
topic_facet |
Ingénierie des aliments Food engineering lipases/acyltransferases;CpLIP2;CAL-A;transesterification;rational design;biocatalysis transesterification acétyltransférase activité enzymatique lipogénique biocatalyse candida parapsilosis lipase |
description |
Performing transesterifications in aqueous media is becoming a priority challenge in lipid biotechnology in order to develop more eco-friendly and efficient biocatalytic processes in systems containing both polar and apolar substrates. In this context, our group has explored for several years the high potential of the lipase/acyltransferase CpLIP2 from Candida parapsilosis and of several of its homologs, that catalyze efficiently acyltransfer reactions in lipid/water media with high water activity (aw > 0.9). The discovery of a new member of this group, CduLAc from C. dubliniensis, with a higher acyltransferase activity than CpLIP2, has provided a new insight on structure-function relationships in this group. Indeed, the comparison of sequences and 3D models, especially of CpLIP2 and CduLAc, with those of the phylogenetically related lipase A from Ps. Antarctica (CAL-A), allowed elucidating a key structural determinant of the acyltransferase activity: serine S369 in CpLIP2 and its equivalents E370 in CAL-A and A366 in CduLAc. Mutants obtained by rational design at this key position showed significant changes in acyltransfer activity. Whereas mutation S369E resulted in an increase in the hydrolytic activity of CpLIP2, S369A increased alcoholysis. More strikingly, the single E370A mutation in CAL-A drastically increased the acyltransferase activity of this enzyme, giving it the character of a lipase/acyltransferase. Indeed, this single mutation lowered the methanol concentration for which the initial rates of alcoholysis and hydrolysis are equal from 2 M in CAL-A down to 0.3 M in its mutant, while the exceptional stability of the parental enzyme toward alcohol and temperature was conserved. |
author2 |
Subileau, Maeva |
format |
Article in Journal/Newspaper |
author |
Jan, Anne Hélène Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric |
author_facet |
Jan, Anne Hélène Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric |
author_sort |
Jan, Anne Hélène |
title |
Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design |
title_short |
Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design |
title_full |
Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design |
title_fullStr |
Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design |
title_full_unstemmed |
Elucidation of a key position for acyltransfer activity in Candida parapsilosis lipase/acyltransferase (CpLIP2) and in pseudozyma Antarctica lipase a (CAL-a) by rational design |
title_sort |
elucidation of a key position for acyltransfer activity in candida parapsilosis lipase/acyltransferase (cplip2) and in pseudozyma antarctica lipase a (cal-a) by rational design |
publishDate |
2016 |
url |
http://prodinra.inra.fr/ft/9CDAC24F-45A6-47B3-9C56-1003E9BC23C7 http://prodinra.inra.fr/record/333999 https://doi.org/10.1016/j.bbapap.2015.11.006 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Biochimica et Biophysica Acta - Proteins and Proteomics 2 (1864), 187–194. (2016) |
op_rights |
http://creativecommons.org/licenses/by-nd-nc/1.0/ |
op_rightsnorm |
CC-BY-ND-NC |
op_doi |
https://doi.org/10.1016/j.bbapap.2015.11.006 |
container_title |
Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics |
container_volume |
1864 |
container_issue |
2 |
container_start_page |
187 |
op_container_end_page |
194 |
_version_ |
1766273718673211392 |