Purification and characterisation of a novel steryl esterase from Melanocarpus albomyces

Hanna Kontkanen (Corresponding Author), Maija Tenkanen, Tapani Reinikainen

Research output: Contribution to journalArticleScientificpeer-review

24 Citations (Scopus)

Abstract

A novel steryl esterase from the ascomycete Melanocarpus albomyces was purified using hydrophobic interaction chromatography and anion exchange chromatography, and the enzyme was biochemically characterised. The enzyme has a tetrameric structure with a molecular weight of 238 kDa. Native gel electrophoresis indicated an isoelectric point of 4.5. The enzyme had broad substrate specificity for different steryl esters, p-nitrophenyl esters and triglycerides with long-chain fatty acids. The pH optimum was dependent on the substrate. The pH optimum of lipase activity was at pH 7, whereas cholesteryl esterase and carboxyl esterase activities had optima at pH 5.5 and 5, respectively. The steryl esterase was more stable at lower pH than at values above pH 7. The enzyme retained over 70% of its activity after 5 h incubation at 50 °C. Activity on p-nitrophenyl butyrate was clearly increased by low detergent concentrations (≤0.1%). M. albomyces steryl esterase is considered to be an interesting tool for the enzymatic modifications of wood pitch due to its broad substrate specificity. Treatment of a model pitch simulating the TMP resin showed that both steryl esters and triglycerides were hydrolysed in the presence of a detergent, whereas only triglycerides were degraded in the absence of detergent.
Original languageEnglish
Pages (from-to)265-273
JournalEnzyme and Microbial Technology
Volume39
Issue number2
DOIs
Publication statusPublished - 2006
MoE publication typeA1 Journal article-refereed

Fingerprint

Esterases
Purification
Detergents
Enzymes
Esters
Triglycerides
Chromatography
Substrate Specificity
Substrates
Thymidine Monophosphate
Lipases
Ascomycota
Electrophoresis
Lipase
Fatty acids
Isoelectric Point
Anions
Ion exchange
Wood
Hydrophobic and Hydrophilic Interactions

Keywords

  • Steryl esterase
  • Lipase
  • Steryl esters
  • Melanocarpus albomyces
  • Purification
  • Characterisation

Cite this

Kontkanen, Hanna ; Tenkanen, Maija ; Reinikainen, Tapani. / Purification and characterisation of a novel steryl esterase from Melanocarpus albomyces. In: Enzyme and Microbial Technology. 2006 ; Vol. 39, No. 2. pp. 265-273.
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abstract = "A novel steryl esterase from the ascomycete Melanocarpus albomyces was purified using hydrophobic interaction chromatography and anion exchange chromatography, and the enzyme was biochemically characterised. The enzyme has a tetrameric structure with a molecular weight of 238 kDa. Native gel electrophoresis indicated an isoelectric point of 4.5. The enzyme had broad substrate specificity for different steryl esters, p-nitrophenyl esters and triglycerides with long-chain fatty acids. The pH optimum was dependent on the substrate. The pH optimum of lipase activity was at pH 7, whereas cholesteryl esterase and carboxyl esterase activities had optima at pH 5.5 and 5, respectively. The steryl esterase was more stable at lower pH than at values above pH 7. The enzyme retained over 70{\%} of its activity after 5 h incubation at 50 °C. Activity on p-nitrophenyl butyrate was clearly increased by low detergent concentrations (≤0.1{\%}). M. albomyces steryl esterase is considered to be an interesting tool for the enzymatic modifications of wood pitch due to its broad substrate specificity. Treatment of a model pitch simulating the TMP resin showed that both steryl esters and triglycerides were hydrolysed in the presence of a detergent, whereas only triglycerides were degraded in the absence of detergent.",
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Purification and characterisation of a novel steryl esterase from Melanocarpus albomyces. / Kontkanen, Hanna (Corresponding Author); Tenkanen, Maija; Reinikainen, Tapani.

In: Enzyme and Microbial Technology, Vol. 39, No. 2, 2006, p. 265-273.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Purification and characterisation of a novel steryl esterase from Melanocarpus albomyces

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AU - Tenkanen, Maija

AU - Reinikainen, Tapani

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N2 - A novel steryl esterase from the ascomycete Melanocarpus albomyces was purified using hydrophobic interaction chromatography and anion exchange chromatography, and the enzyme was biochemically characterised. The enzyme has a tetrameric structure with a molecular weight of 238 kDa. Native gel electrophoresis indicated an isoelectric point of 4.5. The enzyme had broad substrate specificity for different steryl esters, p-nitrophenyl esters and triglycerides with long-chain fatty acids. The pH optimum was dependent on the substrate. The pH optimum of lipase activity was at pH 7, whereas cholesteryl esterase and carboxyl esterase activities had optima at pH 5.5 and 5, respectively. The steryl esterase was more stable at lower pH than at values above pH 7. The enzyme retained over 70% of its activity after 5 h incubation at 50 °C. Activity on p-nitrophenyl butyrate was clearly increased by low detergent concentrations (≤0.1%). M. albomyces steryl esterase is considered to be an interesting tool for the enzymatic modifications of wood pitch due to its broad substrate specificity. Treatment of a model pitch simulating the TMP resin showed that both steryl esters and triglycerides were hydrolysed in the presence of a detergent, whereas only triglycerides were degraded in the absence of detergent.

AB - A novel steryl esterase from the ascomycete Melanocarpus albomyces was purified using hydrophobic interaction chromatography and anion exchange chromatography, and the enzyme was biochemically characterised. The enzyme has a tetrameric structure with a molecular weight of 238 kDa. Native gel electrophoresis indicated an isoelectric point of 4.5. The enzyme had broad substrate specificity for different steryl esters, p-nitrophenyl esters and triglycerides with long-chain fatty acids. The pH optimum was dependent on the substrate. The pH optimum of lipase activity was at pH 7, whereas cholesteryl esterase and carboxyl esterase activities had optima at pH 5.5 and 5, respectively. The steryl esterase was more stable at lower pH than at values above pH 7. The enzyme retained over 70% of its activity after 5 h incubation at 50 °C. Activity on p-nitrophenyl butyrate was clearly increased by low detergent concentrations (≤0.1%). M. albomyces steryl esterase is considered to be an interesting tool for the enzymatic modifications of wood pitch due to its broad substrate specificity. Treatment of a model pitch simulating the TMP resin showed that both steryl esters and triglycerides were hydrolysed in the presence of a detergent, whereas only triglycerides were degraded in the absence of detergent.

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KW - Lipase

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KW - Melanocarpus albomyces

KW - Purification

KW - Characterisation

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