Sol-gel encapsulation: an efficient and versatile immobilization technique for cutinase in non-aqueous media

Pedro Vidinha, Vera Augusto, Miguel Conceição Almeida, Isabel Fonseca, Alexandra Fidalgo, Laura Ilharco, Joaquim M. S. Cabral, Susana Barreiros

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Cutinase from Fusarium solani pisi was encapsulated in sol-gel matrices prepared with a combination of alkyl-alkoxysilane precursors of different chain-lengths. The specific activity of cutinase in a model transesterification reaction at fixed water activity in n-hexane was highest for the precursor combination tetramethoxysilane/n-butyltrimetoxysilane (TMOS/BTMS) in a 1:5 ratio, lower and higher chain lengths of the mono-alkylated precursor or decreasing proportions of the latter relative to TMOS leading to lower enzyme activity. Results obtained using combinations of three precursors confirmed the beneficial effect of the presence of BTMS in the preparations. Scanning electron microscopy of the 1:5 TMOS/n-alkylTMS gels showed a direct correlation between the macropore dimensions and the alkyl chain length of the alkylated precursor and revealed that TMOS/n-octylTMS gels suffered extensive pore collapse during the drying process. The specific activity of TMOS/BTMS sol-gel entrapped cutinase was similar to that exhibited by the enzyme immobilized by adsorption on zeolite NaY. However, the incorporation of different additives (zeolites, silica, Biogel, grinded sol-gel, etc.) having in common the capability to react with residual silanol groups of the sol-gel matrix brought about remarkable enhancements of cutinase activity, despite the fact that the global porosity of the gels did not change. The behavior of the gels in supercritical CO 2 (sc-CO 2 ) paralleled that exhibited in n-hexane, although cutinase activity was ca. one order of magnitude lower (i.e. sol-gel encapsulation did not prevent the deleterious effect of CO 2 ). The impact that functionalization of some of the additives had on cutinase activity indicates that the enzyme/matrix interactions must play an important role. Some of the best additives from the standpoint of enzyme activity were also the best from the standpoint of its operational stability (ca. 80% retention of enzyme activity at the tenth reutilization cycle). None of the additives that proved effective for cutinase could improve the catalytic activity of sol-gel encapsulated Pseudomonas cepacia lipase.

Original languageEnglish
Pages (from-to)23-33
Number of pages11
JournalJournal of Biotechnology
Volume121
Issue number1
DOIs
Publication statusPublished - 2 Jan 2006

Fingerprint

Polymethyl Methacrylate
Encapsulation
Immobilization
Sol-gels
Gels
Enzyme activity
Chain length
Zeolites
Hexane
Carbon Monoxide
Enzymes
Transesterification
Lipases
Immobilized Enzymes
cutinase
Lipase
Catalyst activity
Drying
Silicon Dioxide
Burkholderia cepacia

Keywords

  • Cutinase
  • Organic solvents
  • Pseudomonas cepacia lipase
  • Sol-gel
  • Supercritical fluids
  • Zeolites

Cite this

Vidinha, Pedro ; Augusto, Vera ; Almeida, Miguel Conceição ; Fonseca, Isabel ; Fidalgo, Alexandra ; Ilharco, Laura ; Cabral, Joaquim M. S. ; Barreiros, Susana. / Sol-gel encapsulation: an efficient and versatile immobilization technique for cutinase in non-aqueous media. In: Journal of Biotechnology. 2006 ; Vol. 121, No. 1. pp. 23-33.
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abstract = "Cutinase from Fusarium solani pisi was encapsulated in sol-gel matrices prepared with a combination of alkyl-alkoxysilane precursors of different chain-lengths. The specific activity of cutinase in a model transesterification reaction at fixed water activity in n-hexane was highest for the precursor combination tetramethoxysilane/n-butyltrimetoxysilane (TMOS/BTMS) in a 1:5 ratio, lower and higher chain lengths of the mono-alkylated precursor or decreasing proportions of the latter relative to TMOS leading to lower enzyme activity. Results obtained using combinations of three precursors confirmed the beneficial effect of the presence of BTMS in the preparations. Scanning electron microscopy of the 1:5 TMOS/n-alkylTMS gels showed a direct correlation between the macropore dimensions and the alkyl chain length of the alkylated precursor and revealed that TMOS/n-octylTMS gels suffered extensive pore collapse during the drying process. The specific activity of TMOS/BTMS sol-gel entrapped cutinase was similar to that exhibited by the enzyme immobilized by adsorption on zeolite NaY. However, the incorporation of different additives (zeolites, silica, Biogel, grinded sol-gel, etc.) having in common the capability to react with residual silanol groups of the sol-gel matrix brought about remarkable enhancements of cutinase activity, despite the fact that the global porosity of the gels did not change. The behavior of the gels in supercritical CO 2 (sc-CO 2 ) paralleled that exhibited in n-hexane, although cutinase activity was ca. one order of magnitude lower (i.e. sol-gel encapsulation did not prevent the deleterious effect of CO 2 ). The impact that functionalization of some of the additives had on cutinase activity indicates that the enzyme/matrix interactions must play an important role. Some of the best additives from the standpoint of enzyme activity were also the best from the standpoint of its operational stability (ca. 80{\%} retention of enzyme activity at the tenth reutilization cycle). None of the additives that proved effective for cutinase could improve the catalytic activity of sol-gel encapsulated Pseudomonas cepacia lipase.",
keywords = "Cutinase, Organic solvents, Pseudomonas cepacia lipase, Sol-gel, Supercritical fluids, Zeolites",
author = "Pedro Vidinha and Vera Augusto and Almeida, {Miguel Concei{\cc}{\~a}o} and Isabel Fonseca and Alexandra Fidalgo and Laura Ilharco and Cabral, {Joaquim M. S.} and Susana Barreiros",
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Sol-gel encapsulation: an efficient and versatile immobilization technique for cutinase in non-aqueous media. / Vidinha, Pedro; Augusto, Vera; Almeida, Miguel Conceição; Fonseca, Isabel; Fidalgo, Alexandra; Ilharco, Laura; Cabral, Joaquim M. S.; Barreiros, Susana.

In: Journal of Biotechnology, Vol. 121, No. 1, 02.01.2006, p. 23-33.

Research output: Contribution to journalArticle

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T1 - Sol-gel encapsulation: an efficient and versatile immobilization technique for cutinase in non-aqueous media

AU - Vidinha, Pedro

AU - Augusto, Vera

AU - Almeida, Miguel Conceição

AU - Fonseca, Isabel

AU - Fidalgo, Alexandra

AU - Ilharco, Laura

AU - Cabral, Joaquim M. S.

AU - Barreiros, Susana

N1 - This work has been supported by Fundação para a Ciência e Tecnologia (FCT, Portugal) through the contract POCTI/35429/QUI/2000 and the grant PRAXIS XXI/SFRH/BD/2003 (P. Vidinha), and by FEDER.

PY - 2006/1/2

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N2 - Cutinase from Fusarium solani pisi was encapsulated in sol-gel matrices prepared with a combination of alkyl-alkoxysilane precursors of different chain-lengths. The specific activity of cutinase in a model transesterification reaction at fixed water activity in n-hexane was highest for the precursor combination tetramethoxysilane/n-butyltrimetoxysilane (TMOS/BTMS) in a 1:5 ratio, lower and higher chain lengths of the mono-alkylated precursor or decreasing proportions of the latter relative to TMOS leading to lower enzyme activity. Results obtained using combinations of three precursors confirmed the beneficial effect of the presence of BTMS in the preparations. Scanning electron microscopy of the 1:5 TMOS/n-alkylTMS gels showed a direct correlation between the macropore dimensions and the alkyl chain length of the alkylated precursor and revealed that TMOS/n-octylTMS gels suffered extensive pore collapse during the drying process. The specific activity of TMOS/BTMS sol-gel entrapped cutinase was similar to that exhibited by the enzyme immobilized by adsorption on zeolite NaY. However, the incorporation of different additives (zeolites, silica, Biogel, grinded sol-gel, etc.) having in common the capability to react with residual silanol groups of the sol-gel matrix brought about remarkable enhancements of cutinase activity, despite the fact that the global porosity of the gels did not change. The behavior of the gels in supercritical CO 2 (sc-CO 2 ) paralleled that exhibited in n-hexane, although cutinase activity was ca. one order of magnitude lower (i.e. sol-gel encapsulation did not prevent the deleterious effect of CO 2 ). The impact that functionalization of some of the additives had on cutinase activity indicates that the enzyme/matrix interactions must play an important role. Some of the best additives from the standpoint of enzyme activity were also the best from the standpoint of its operational stability (ca. 80% retention of enzyme activity at the tenth reutilization cycle). None of the additives that proved effective for cutinase could improve the catalytic activity of sol-gel encapsulated Pseudomonas cepacia lipase.

AB - Cutinase from Fusarium solani pisi was encapsulated in sol-gel matrices prepared with a combination of alkyl-alkoxysilane precursors of different chain-lengths. The specific activity of cutinase in a model transesterification reaction at fixed water activity in n-hexane was highest for the precursor combination tetramethoxysilane/n-butyltrimetoxysilane (TMOS/BTMS) in a 1:5 ratio, lower and higher chain lengths of the mono-alkylated precursor or decreasing proportions of the latter relative to TMOS leading to lower enzyme activity. Results obtained using combinations of three precursors confirmed the beneficial effect of the presence of BTMS in the preparations. Scanning electron microscopy of the 1:5 TMOS/n-alkylTMS gels showed a direct correlation between the macropore dimensions and the alkyl chain length of the alkylated precursor and revealed that TMOS/n-octylTMS gels suffered extensive pore collapse during the drying process. The specific activity of TMOS/BTMS sol-gel entrapped cutinase was similar to that exhibited by the enzyme immobilized by adsorption on zeolite NaY. However, the incorporation of different additives (zeolites, silica, Biogel, grinded sol-gel, etc.) having in common the capability to react with residual silanol groups of the sol-gel matrix brought about remarkable enhancements of cutinase activity, despite the fact that the global porosity of the gels did not change. The behavior of the gels in supercritical CO 2 (sc-CO 2 ) paralleled that exhibited in n-hexane, although cutinase activity was ca. one order of magnitude lower (i.e. sol-gel encapsulation did not prevent the deleterious effect of CO 2 ). The impact that functionalization of some of the additives had on cutinase activity indicates that the enzyme/matrix interactions must play an important role. Some of the best additives from the standpoint of enzyme activity were also the best from the standpoint of its operational stability (ca. 80% retention of enzyme activity at the tenth reutilization cycle). None of the additives that proved effective for cutinase could improve the catalytic activity of sol-gel encapsulated Pseudomonas cepacia lipase.

KW - Cutinase

KW - Organic solvents

KW - Pseudomonas cepacia lipase

KW - Sol-gel

KW - Supercritical fluids

KW - Zeolites

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