An organic solvent-tolerant lipase with both hydrolytic and synthetic activities from the oleaginous fungus mortierella echinosphaera

Alexandra Kotogán, Carolina Zambrano, Anita Kecskeméti, Mónika Varga, A. Szekeres, T. Papp, C. Vágvölgyi, Miklós Takó

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Lipase enzymes of the oleaginous fungal group Mortierella are rarely studied. However, considering that most commercial lipases are derived from filamentous fungal sources, their investigation can contribute to the cost-effective development of new biotechnological processes. Here, an extracellular lipase with a molecular mass of 30 kDa was isolated from Mortierella echinosphaera CBS 575.75 and characterized. The purified lipase exhibited an optimal p-nitrophenyl palmitate (pNPP)-hydrolyzing activity at 25C and pH 6.6–7.0 and proved to be highly stable at temperatures up to 40C and under broad pH conditions. The enzyme was active under low temperatures, retaining 32.5% of its activity at 10C, and was significantly stable in polar and non-polar organic solvents. The Km, Vmax, and kcat for pNPP were 0.336 mM, 30.4 µM/min, and 45.7 1/min for pNPP and 0.333 mM, 36.9 µM/min, and 55.6 1/min for pNP-decanoate, respectively. The pNPP hydrolysis was inhibited by Hg2+, N-bromosuccinimide, and sodium dodecyl sulfate, while ethylenediaminetetraacetic acid and metal ions, such as Ca2+, Mg2+, Na+, and K+ enhanced the activity. The purified lipase had non-regioselective activity and wide substrate specificity, showing a clear preference for medium-chained p-nitrophenyl esters. Besides its good transesterification activity, the enzyme appeared as a suitable biocatalyst to operate selective esterification reactions to long-chained alkyl esters. Adsorption to Accurel MP1000 improved the storage stability of the enzyme at 5C. The immobilized lipase displayed tolerance to a non-aqueous environment and was reusable for up to five cycles without significant loss in its synthetic and hydrolytic activities. These findings confirm the applicability of both the free and the immobilized enzyme preparations in future research.

Original languageEnglish
Article number1129
JournalInternational Journal of Molecular Sciences
Volume19
Issue number4
DOIs
Publication statusPublished - Apr 10 2018

Fingerprint

Mortierella
fungi
Lipases
Fungi
Lipase
Organic solvents
Palmitates
enzymes
Enzymes
esters
Esters
Bromosuccinimide
storage stability
Decanoates
Enzyme Stability
Immobilized Enzymes
Biocatalysts
Temperature
ethylenediaminetetraacetic acids
Esterification

Keywords

  • Alkyl esters
  • Biochemical characterization
  • Esterification
  • Extracellular lipase
  • Immobilization
  • P-nitrophenyl esters
  • Transesterification
  • Zygomycetes

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

Cite this

An organic solvent-tolerant lipase with both hydrolytic and synthetic activities from the oleaginous fungus mortierella echinosphaera. / Kotogán, Alexandra; Zambrano, Carolina; Kecskeméti, Anita; Varga, Mónika; Szekeres, A.; Papp, T.; Vágvölgyi, C.; Takó, Miklós.

In: International Journal of Molecular Sciences, Vol. 19, No. 4, 1129, 10.04.2018.

Research output: Contribution to journalArticle

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abstract = "Lipase enzymes of the oleaginous fungal group Mortierella are rarely studied. However, considering that most commercial lipases are derived from filamentous fungal sources, their investigation can contribute to the cost-effective development of new biotechnological processes. Here, an extracellular lipase with a molecular mass of 30 kDa was isolated from Mortierella echinosphaera CBS 575.75 and characterized. The purified lipase exhibited an optimal p-nitrophenyl palmitate (pNPP)-hydrolyzing activity at 25◦C and pH 6.6–7.0 and proved to be highly stable at temperatures up to 40◦C and under broad pH conditions. The enzyme was active under low temperatures, retaining 32.5{\%} of its activity at 10◦C, and was significantly stable in polar and non-polar organic solvents. The Km, Vmax, and kcat for pNPP were 0.336 mM, 30.4 µM/min, and 45.7 1/min for pNPP and 0.333 mM, 36.9 µM/min, and 55.6 1/min for pNP-decanoate, respectively. The pNPP hydrolysis was inhibited by Hg2+, N-bromosuccinimide, and sodium dodecyl sulfate, while ethylenediaminetetraacetic acid and metal ions, such as Ca2+, Mg2+, Na+, and K+ enhanced the activity. The purified lipase had non-regioselective activity and wide substrate specificity, showing a clear preference for medium-chained p-nitrophenyl esters. Besides its good transesterification activity, the enzyme appeared as a suitable biocatalyst to operate selective esterification reactions to long-chained alkyl esters. Adsorption to Accurel MP1000 improved the storage stability of the enzyme at 5◦C. The immobilized lipase displayed tolerance to a non-aqueous environment and was reusable for up to five cycles without significant loss in its synthetic and hydrolytic activities. These findings confirm the applicability of both the free and the immobilized enzyme preparations in future research.",
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AU - Kotogán, Alexandra

AU - Zambrano, Carolina

AU - Kecskeméti, Anita

AU - Varga, Mónika

AU - Szekeres, A.

AU - Papp, T.

AU - Vágvölgyi, C.

AU - Takó, Miklós

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N2 - Lipase enzymes of the oleaginous fungal group Mortierella are rarely studied. However, considering that most commercial lipases are derived from filamentous fungal sources, their investigation can contribute to the cost-effective development of new biotechnological processes. Here, an extracellular lipase with a molecular mass of 30 kDa was isolated from Mortierella echinosphaera CBS 575.75 and characterized. The purified lipase exhibited an optimal p-nitrophenyl palmitate (pNPP)-hydrolyzing activity at 25◦C and pH 6.6–7.0 and proved to be highly stable at temperatures up to 40◦C and under broad pH conditions. The enzyme was active under low temperatures, retaining 32.5% of its activity at 10◦C, and was significantly stable in polar and non-polar organic solvents. The Km, Vmax, and kcat for pNPP were 0.336 mM, 30.4 µM/min, and 45.7 1/min for pNPP and 0.333 mM, 36.9 µM/min, and 55.6 1/min for pNP-decanoate, respectively. The pNPP hydrolysis was inhibited by Hg2+, N-bromosuccinimide, and sodium dodecyl sulfate, while ethylenediaminetetraacetic acid and metal ions, such as Ca2+, Mg2+, Na+, and K+ enhanced the activity. The purified lipase had non-regioselective activity and wide substrate specificity, showing a clear preference for medium-chained p-nitrophenyl esters. Besides its good transesterification activity, the enzyme appeared as a suitable biocatalyst to operate selective esterification reactions to long-chained alkyl esters. Adsorption to Accurel MP1000 improved the storage stability of the enzyme at 5◦C. The immobilized lipase displayed tolerance to a non-aqueous environment and was reusable for up to five cycles without significant loss in its synthetic and hydrolytic activities. These findings confirm the applicability of both the free and the immobilized enzyme preparations in future research.

AB - Lipase enzymes of the oleaginous fungal group Mortierella are rarely studied. However, considering that most commercial lipases are derived from filamentous fungal sources, their investigation can contribute to the cost-effective development of new biotechnological processes. Here, an extracellular lipase with a molecular mass of 30 kDa was isolated from Mortierella echinosphaera CBS 575.75 and characterized. The purified lipase exhibited an optimal p-nitrophenyl palmitate (pNPP)-hydrolyzing activity at 25◦C and pH 6.6–7.0 and proved to be highly stable at temperatures up to 40◦C and under broad pH conditions. The enzyme was active under low temperatures, retaining 32.5% of its activity at 10◦C, and was significantly stable in polar and non-polar organic solvents. The Km, Vmax, and kcat for pNPP were 0.336 mM, 30.4 µM/min, and 45.7 1/min for pNPP and 0.333 mM, 36.9 µM/min, and 55.6 1/min for pNP-decanoate, respectively. The pNPP hydrolysis was inhibited by Hg2+, N-bromosuccinimide, and sodium dodecyl sulfate, while ethylenediaminetetraacetic acid and metal ions, such as Ca2+, Mg2+, Na+, and K+ enhanced the activity. The purified lipase had non-regioselective activity and wide substrate specificity, showing a clear preference for medium-chained p-nitrophenyl esters. Besides its good transesterification activity, the enzyme appeared as a suitable biocatalyst to operate selective esterification reactions to long-chained alkyl esters. Adsorption to Accurel MP1000 improved the storage stability of the enzyme at 5◦C. The immobilized lipase displayed tolerance to a non-aqueous environment and was reusable for up to five cycles without significant loss in its synthetic and hydrolytic activities. These findings confirm the applicability of both the free and the immobilized enzyme preparations in future research.

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KW - Transesterification

KW - Zygomycetes

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