Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] 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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Online Access: | https://hal.science/hal-01269389 https://hal.science/hal-01269389/document https://hal.science/hal-01269389/file/Jan-BBA-2016-manuscript_1.pdf https://doi.org/10.1016/j.bbapap.2015.11.006 |
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ftsupagro:oai:HAL:hal-01269389v1 2023-07-30T03:58:40+02:00 Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] lipase a (CAL-a) by rational design Jan, Anne Hélène Subileau, Maeva Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE) Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro) Metaglyc 2 project (no. 22008910) BASF 2016 https://hal.science/hal-01269389 https://hal.science/hal-01269389/document https://hal.science/hal-01269389/file/Jan-BBA-2016-manuscript_1.pdf https://doi.org/10.1016/j.bbapap.2015.11.006 en eng HAL CCSD Elsevier info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbapap.2015.11.006 hal-01269389 https://hal.science/hal-01269389 https://hal.science/hal-01269389/document https://hal.science/hal-01269389/file/Jan-BBA-2016-manuscript_1.pdf doi:10.1016/j.bbapap.2015.11.006 PRODINRA: 333999 WOS: 000368566000001 info:eu-repo/semantics/OpenAccess ISSN: 1570-9639 Biochimica et Biophysica Acta Proteins and Proteomics https://hal.science/hal-01269389 Biochimica et Biophysica Acta Proteins and Proteomics, 2016, 1864 (2), pp.187-194. ⟨10.1016/j.bbapap.2015.11.006⟩ lipases/acyltransferases CpLIP2 CAL-A transesterification rational design biocatalysis [SDV.IDA]Life Sciences [q-bio]/Food engineering info:eu-repo/semantics/article Journal articles 2016 ftsupagro https://doi.org/10.1016/j.bbapap.2015.11.006 2023-07-08T08:53:18Z 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 Unknown Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics 1864 2 187 194 |
institution |
Open Polar |
collection |
Unknown |
op_collection_id |
ftsupagro |
language |
English |
topic |
lipases/acyltransferases CpLIP2 CAL-A transesterification rational design biocatalysis [SDV.IDA]Life Sciences [q-bio]/Food engineering |
spellingShingle |
lipases/acyltransferases CpLIP2 CAL-A transesterification rational design biocatalysis [SDV.IDA]Life Sciences [q-bio]/Food engineering Jan, Anne Hélène Subileau, Maeva Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] lipase a (CAL-a) by rational design |
topic_facet |
lipases/acyltransferases CpLIP2 CAL-A transesterification rational design biocatalysis [SDV.IDA]Life Sciences [q-bio]/Food engineering |
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 |
Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE) Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro) Metaglyc 2 project (no. 22008910) BASF |
format |
Article in Journal/Newspaper |
author |
Jan, Anne Hélène Subileau, Maeva Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric |
author_facet |
Jan, Anne Hélène Subileau, Maeva Deyrieux, Charlotte Perrier, Véronique Dubreucq, Eric |
author_sort |
Jan, Anne Hélène |
title |
Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] lipase a (CAL-a) by rational design |
title_short |
Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] lipase a (CAL-a) by rational design |
title_full |
Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] lipase a (CAL-a) by rational design |
title_fullStr |
Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] lipase a (CAL-a) by rational design |
title_full_unstemmed |
Elucidation of a key position for acyltransfer activity in[i] Candida parapsilosis[/i] lipase/acyltransferase (CpLIP2) and in [i]pseudozyma Antarctica[/i] lipase a (CAL-a) by rational design |
title_sort |
elucidation of a key position for acyltransfer activity in[i] candida parapsilosis[/i] lipase/acyltransferase (cplip2) and in [i]pseudozyma antarctica[/i] lipase a (cal-a) by rational design |
publisher |
HAL CCSD |
publishDate |
2016 |
url |
https://hal.science/hal-01269389 https://hal.science/hal-01269389/document https://hal.science/hal-01269389/file/Jan-BBA-2016-manuscript_1.pdf https://doi.org/10.1016/j.bbapap.2015.11.006 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
ISSN: 1570-9639 Biochimica et Biophysica Acta Proteins and Proteomics https://hal.science/hal-01269389 Biochimica et Biophysica Acta Proteins and Proteomics, 2016, 1864 (2), pp.187-194. ⟨10.1016/j.bbapap.2015.11.006⟩ |
op_relation |
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbapap.2015.11.006 hal-01269389 https://hal.science/hal-01269389 https://hal.science/hal-01269389/document https://hal.science/hal-01269389/file/Jan-BBA-2016-manuscript_1.pdf doi:10.1016/j.bbapap.2015.11.006 PRODINRA: 333999 WOS: 000368566000001 |
op_rights |
info:eu-repo/semantics/OpenAccess |
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 |
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1772821419409801216 |