Understanding the silica-based sol-gel encapsulation mechanism of Thermomyces lanuginosus lipase: The role of polyethylenimine

Sindy Escobar, Claudia Bernal, Juan M. Bolivar, Bernd Nidetzky, Fernando López-Gallego, Monica Mesa

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Silica-based sol-gel enzyme immobilization aided by polyamines has been used for generating active and stable heterogeneous biocatalysts. However, the encapsulation mechanism and its influence on the characteristics of the resulting biocatalysts have not been fully understood yet. This work aimed to shed light on how Thermomyces lanuginosus lipase, siliceous species and the additive polyethylenimine (PEI) interact during enzyme encapsulation, by establishing a correlation between addition of PEI, material formation and enzyme encapsulation. The addition of PEI appears to increase enzyme incorporation producing a biocatalyst with high encapsulation efficiency in terms of percentage of encapsulated protein (90%) and activity (47%). Furthermore, we observed that the addition of PEI resulted in a biocatalyst with high catalytic activity (600 U/g) and thermal stability (247-fold more stable than the soluble enzyme at 65 °C). The formation of silica particles was followed over time, showing a homogeneous incorporation of the enzyme in its active form. Intrinsic fluorescence spectroscopy suggested that the addition of PEI during the encapsulation process resulted in a conservation of the structural and functional features of the enzyme, suggesting a stabilizing role of the additive. The integration of the results of this work allowed proposing a polycondensation/enzyme encapsulation mechanism for silica-based sol-gel encapsulation mediated by PEI. These new evidences will contribute to the design of novel and more efficient silica-based biocatalysts achieved by sol gel encapsulation methodology.

Original languageEnglish
Pages (from-to)106-113
Number of pages8
JournalMolecular Catalysis
DOIs
Publication statusPublished - 1 Apr 2018

Fingerprint

Polyethyleneimine
Lipases
Lipase
Encapsulation
Silicon Dioxide
Sol-gels
enzymes
Biocatalysts
Silica
gels
silicon dioxide
Enzymes
Enzyme immobilization
Fluorescence spectroscopy
Polycondensation
immobilization
catalytic activity
conservation
Catalyst activity
Conservation

Keywords

  • Encapsulation
  • Polyethylenimine
  • Sol-gel process
  • Thermomyces lanuginosus lipase

ASJC Scopus subject areas

  • Catalysis
  • Process Chemistry and Technology
  • Physical and Theoretical Chemistry

Cite this

Understanding the silica-based sol-gel encapsulation mechanism of Thermomyces lanuginosus lipase : The role of polyethylenimine. / Escobar, Sindy; Bernal, Claudia; Bolivar, Juan M.; Nidetzky, Bernd; López-Gallego, Fernando; Mesa, Monica.

In: Molecular Catalysis, 01.04.2018, p. 106-113.

Research output: Contribution to journalArticleResearchpeer-review

Escobar, Sindy ; Bernal, Claudia ; Bolivar, Juan M. ; Nidetzky, Bernd ; López-Gallego, Fernando ; Mesa, Monica. / Understanding the silica-based sol-gel encapsulation mechanism of Thermomyces lanuginosus lipase : The role of polyethylenimine. In: Molecular Catalysis. 2018 ; pp. 106-113.
@article{ad113e7fc86943588e7f4901585cff8b,
title = "Understanding the silica-based sol-gel encapsulation mechanism of Thermomyces lanuginosus lipase: The role of polyethylenimine",
abstract = "Silica-based sol-gel enzyme immobilization aided by polyamines has been used for generating active and stable heterogeneous biocatalysts. However, the encapsulation mechanism and its influence on the characteristics of the resulting biocatalysts have not been fully understood yet. This work aimed to shed light on how Thermomyces lanuginosus lipase, siliceous species and the additive polyethylenimine (PEI) interact during enzyme encapsulation, by establishing a correlation between addition of PEI, material formation and enzyme encapsulation. The addition of PEI appears to increase enzyme incorporation producing a biocatalyst with high encapsulation efficiency in terms of percentage of encapsulated protein (90{\%}) and activity (47{\%}). Furthermore, we observed that the addition of PEI resulted in a biocatalyst with high catalytic activity (600 U/g) and thermal stability (247-fold more stable than the soluble enzyme at 65 °C). The formation of silica particles was followed over time, showing a homogeneous incorporation of the enzyme in its active form. Intrinsic fluorescence spectroscopy suggested that the addition of PEI during the encapsulation process resulted in a conservation of the structural and functional features of the enzyme, suggesting a stabilizing role of the additive. The integration of the results of this work allowed proposing a polycondensation/enzyme encapsulation mechanism for silica-based sol-gel encapsulation mediated by PEI. These new evidences will contribute to the design of novel and more efficient silica-based biocatalysts achieved by sol gel encapsulation methodology.",
keywords = "Encapsulation, Polyethylenimine, Sol-gel process, Thermomyces lanuginosus lipase",
author = "Sindy Escobar and Claudia Bernal and Bolivar, {Juan M.} and Bernd Nidetzky and Fernando L{\'o}pez-Gallego and Monica Mesa",
year = "2018",
month = "4",
day = "1",
doi = "10.1016/j.mcat.2018.02.024",
language = "English",
pages = "106--113",
journal = "Molecular Catalysis",
issn = "2468-8231",
publisher = "Elsevier B.V.",

}

TY - JOUR

T1 - Understanding the silica-based sol-gel encapsulation mechanism of Thermomyces lanuginosus lipase

T2 - The role of polyethylenimine

AU - Escobar, Sindy

AU - Bernal, Claudia

AU - Bolivar, Juan M.

AU - Nidetzky, Bernd

AU - López-Gallego, Fernando

AU - Mesa, Monica

PY - 2018/4/1

Y1 - 2018/4/1

N2 - Silica-based sol-gel enzyme immobilization aided by polyamines has been used for generating active and stable heterogeneous biocatalysts. However, the encapsulation mechanism and its influence on the characteristics of the resulting biocatalysts have not been fully understood yet. This work aimed to shed light on how Thermomyces lanuginosus lipase, siliceous species and the additive polyethylenimine (PEI) interact during enzyme encapsulation, by establishing a correlation between addition of PEI, material formation and enzyme encapsulation. The addition of PEI appears to increase enzyme incorporation producing a biocatalyst with high encapsulation efficiency in terms of percentage of encapsulated protein (90%) and activity (47%). Furthermore, we observed that the addition of PEI resulted in a biocatalyst with high catalytic activity (600 U/g) and thermal stability (247-fold more stable than the soluble enzyme at 65 °C). The formation of silica particles was followed over time, showing a homogeneous incorporation of the enzyme in its active form. Intrinsic fluorescence spectroscopy suggested that the addition of PEI during the encapsulation process resulted in a conservation of the structural and functional features of the enzyme, suggesting a stabilizing role of the additive. The integration of the results of this work allowed proposing a polycondensation/enzyme encapsulation mechanism for silica-based sol-gel encapsulation mediated by PEI. These new evidences will contribute to the design of novel and more efficient silica-based biocatalysts achieved by sol gel encapsulation methodology.

AB - Silica-based sol-gel enzyme immobilization aided by polyamines has been used for generating active and stable heterogeneous biocatalysts. However, the encapsulation mechanism and its influence on the characteristics of the resulting biocatalysts have not been fully understood yet. This work aimed to shed light on how Thermomyces lanuginosus lipase, siliceous species and the additive polyethylenimine (PEI) interact during enzyme encapsulation, by establishing a correlation between addition of PEI, material formation and enzyme encapsulation. The addition of PEI appears to increase enzyme incorporation producing a biocatalyst with high encapsulation efficiency in terms of percentage of encapsulated protein (90%) and activity (47%). Furthermore, we observed that the addition of PEI resulted in a biocatalyst with high catalytic activity (600 U/g) and thermal stability (247-fold more stable than the soluble enzyme at 65 °C). The formation of silica particles was followed over time, showing a homogeneous incorporation of the enzyme in its active form. Intrinsic fluorescence spectroscopy suggested that the addition of PEI during the encapsulation process resulted in a conservation of the structural and functional features of the enzyme, suggesting a stabilizing role of the additive. The integration of the results of this work allowed proposing a polycondensation/enzyme encapsulation mechanism for silica-based sol-gel encapsulation mediated by PEI. These new evidences will contribute to the design of novel and more efficient silica-based biocatalysts achieved by sol gel encapsulation methodology.

KW - Encapsulation

KW - Polyethylenimine

KW - Sol-gel process

KW - Thermomyces lanuginosus lipase

UR - http://www.scopus.com/inward/record.url?scp=85056421921&partnerID=8YFLogxK

U2 - 10.1016/j.mcat.2018.02.024

DO - 10.1016/j.mcat.2018.02.024

M3 - Article

SP - 106

EP - 113

JO - Molecular Catalysis

JF - Molecular Catalysis

SN - 2468-8231

ER -