Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones
The lipases A and B from Candida antarctica (CALA and CALB), Thermomyces lanuginosus (TLL) or Rhizomucor miehei (RML), and the commercial and artificial phospholipase Lecitase ultra (LEU) may be co-immobilized on octyl agarose beads. However, LEU and RML became almost fully inactivated under conditi...
Published in: | Catalysts |
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Language: | English |
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MDPI AG
2020
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Online Access: | https://doi.org/10.3390/catal10101207 https://doaj.org/article/60bfbced70d54e47a8ded29eebd1bddf |
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ftdoajarticles:oai:doaj.org/article:60bfbced70d54e47a8ded29eebd1bddf 2023-05-15T14:00:35+02:00 Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones Sara Arana-Peña Diego Carballares Vicente Cortés Corberan Roberto Fernandez-Lafuente 2020-10-01T00:00:00Z https://doi.org/10.3390/catal10101207 https://doaj.org/article/60bfbced70d54e47a8ded29eebd1bddf EN eng MDPI AG https://www.mdpi.com/2073-4344/10/10/1207 https://doaj.org/toc/2073-4344 doi:10.3390/catal10101207 2073-4344 https://doaj.org/article/60bfbced70d54e47a8ded29eebd1bddf Catalysts, Vol 10, Iss 1207, p 1207 (2020) combilipase enzyme co-immobilization reuse of the most stable co-immobilized enzyme interfacial activation PEI in enzyme immobilization Chemical technology TP1-1185 Chemistry QD1-999 article 2020 ftdoajarticles https://doi.org/10.3390/catal10101207 2022-12-31T12:52:55Z The lipases A and B from Candida antarctica (CALA and CALB), Thermomyces lanuginosus (TLL) or Rhizomucor miehei (RML), and the commercial and artificial phospholipase Lecitase ultra (LEU) may be co-immobilized on octyl agarose beads. However, LEU and RML became almost fully inactivated under conditions where CALA, CALB and TLL retained full activity. This means that, to have a five components co-immobilized combi-lipase, we should discard 3 fully active and immobilized enzymes when the other two enzymes are inactivated. To solve this situation, CALA, CALB and TLL have been co-immobilized on octyl-vinyl sulfone agarose beads, coated with polyethylenimine (PEI) and the least stable enzymes, RML and LEU have been co-immobilized over these immobilized enzymes. The coating with PEI is even favorable for the activity of the immobilized enzymes. It was checked that RML and LEU could be released from the enzyme-PEI coated biocatalyst, although this also produced some release of the PEI. That way, a protocol was developed to co-immobilize the five enzymes, in a way that the most stable could be reused after the inactivation of the least stable ones. After RML and LEU inactivation, the combi-biocatalysts were incubated in 0.5 M of ammonium sulfate to release the inactivated enzymes, incubated again with PEI and a new RML and LEU batch could be immobilized, maintaining the activity of the three most stable enzymes for at least five cycles of incubation at pH 7.0 and 60 °C for 3 h, incubation on ammonium sulfate, incubation in PEI and co-immobilization of new enzymes. The effect of the order of co-immobilization of the different enzymes on the co-immobilized biocatalyst activity was also investigated using different substrates, finding that when the most active enzyme versus one substrate was immobilized first (nearer to the surface of the particle), the activity was higher than when this enzyme was co-immobilized last (nearer to the particle core). Article in Journal/Newspaper Antarc* Antarctica Directory of Open Access Journals: DOAJ Articles Catalysts 10 10 1207 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
combilipase enzyme co-immobilization reuse of the most stable co-immobilized enzyme interfacial activation PEI in enzyme immobilization Chemical technology TP1-1185 Chemistry QD1-999 |
spellingShingle |
combilipase enzyme co-immobilization reuse of the most stable co-immobilized enzyme interfacial activation PEI in enzyme immobilization Chemical technology TP1-1185 Chemistry QD1-999 Sara Arana-Peña Diego Carballares Vicente Cortés Corberan Roberto Fernandez-Lafuente Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones |
topic_facet |
combilipase enzyme co-immobilization reuse of the most stable co-immobilized enzyme interfacial activation PEI in enzyme immobilization Chemical technology TP1-1185 Chemistry QD1-999 |
description |
The lipases A and B from Candida antarctica (CALA and CALB), Thermomyces lanuginosus (TLL) or Rhizomucor miehei (RML), and the commercial and artificial phospholipase Lecitase ultra (LEU) may be co-immobilized on octyl agarose beads. However, LEU and RML became almost fully inactivated under conditions where CALA, CALB and TLL retained full activity. This means that, to have a five components co-immobilized combi-lipase, we should discard 3 fully active and immobilized enzymes when the other two enzymes are inactivated. To solve this situation, CALA, CALB and TLL have been co-immobilized on octyl-vinyl sulfone agarose beads, coated with polyethylenimine (PEI) and the least stable enzymes, RML and LEU have been co-immobilized over these immobilized enzymes. The coating with PEI is even favorable for the activity of the immobilized enzymes. It was checked that RML and LEU could be released from the enzyme-PEI coated biocatalyst, although this also produced some release of the PEI. That way, a protocol was developed to co-immobilize the five enzymes, in a way that the most stable could be reused after the inactivation of the least stable ones. After RML and LEU inactivation, the combi-biocatalysts were incubated in 0.5 M of ammonium sulfate to release the inactivated enzymes, incubated again with PEI and a new RML and LEU batch could be immobilized, maintaining the activity of the three most stable enzymes for at least five cycles of incubation at pH 7.0 and 60 °C for 3 h, incubation on ammonium sulfate, incubation in PEI and co-immobilization of new enzymes. The effect of the order of co-immobilization of the different enzymes on the co-immobilized biocatalyst activity was also investigated using different substrates, finding that when the most active enzyme versus one substrate was immobilized first (nearer to the surface of the particle), the activity was higher than when this enzyme was co-immobilized last (nearer to the particle core). |
format |
Article in Journal/Newspaper |
author |
Sara Arana-Peña Diego Carballares Vicente Cortés Corberan Roberto Fernandez-Lafuente |
author_facet |
Sara Arana-Peña Diego Carballares Vicente Cortés Corberan Roberto Fernandez-Lafuente |
author_sort |
Sara Arana-Peña |
title |
Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones |
title_short |
Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones |
title_full |
Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones |
title_fullStr |
Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones |
title_full_unstemmed |
Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones |
title_sort |
multi-combilipases: co-immobilizing lipases with very different stabilities combining immobilization via interfacial activation and ion exchange. the reuse of the most stable co-immobilized enzymes after inactivation of the least stable ones |
publisher |
MDPI AG |
publishDate |
2020 |
url |
https://doi.org/10.3390/catal10101207 https://doaj.org/article/60bfbced70d54e47a8ded29eebd1bddf |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Catalysts, Vol 10, Iss 1207, p 1207 (2020) |
op_relation |
https://www.mdpi.com/2073-4344/10/10/1207 https://doaj.org/toc/2073-4344 doi:10.3390/catal10101207 2073-4344 https://doaj.org/article/60bfbced70d54e47a8ded29eebd1bddf |
op_doi |
https://doi.org/10.3390/catal10101207 |
container_title |
Catalysts |
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10 |
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10 |
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1207 |
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