Purification and properties of Amycolatopsis mediterranei DSM 43304 lipase and its potential in flavour ester synthesis

Dharmendra S. Dheeman; Gary T.M. Henehan; Jesus M. Frías

doi:10.1016/j.biortech.2010.11.074

Purification and properties of Amycolatopsis mediterranei DSM 43304 lipase and its potential in flavour ester synthesis

Dharmendra S. Dheeman, Gary T.M. Henehan, Jesus M. Frías

School of Food Science and Environmental Health

Research output: Contribution to journal › Article › peer-review

46 Citations (Scopus)

Abstract

An extracellular thermostable lipase from Amycolatopsis mediterranei DSM 43304 has been purified to homogeneity using ammonium sulphate precipitation followed by anion exchange chromatography and hydrophobic interaction chromatography. This protocol resulted in a 398-fold purification with 36% final recovery. The purified A. mediterranei DSM 43304 lipase (AML) has an apparent molecular mass of 33 kDa. The N-terminal sequence, AANPYERGPDPTTASIEATR, showed highest similarity to a lipase from Streptomyces exfoliatus. The values of K_m^app and V_max^app for p-nitrophenyl palmitate (p-NPP) at the optimal temperature (60. °C) and pH (8.0) were 0.099. ±. 0.010 mM and 2.53. ±. 0.06 mmol/min. mg, respectively. The purified AML displayed significant activity towards a range of short and long chain triglyceride substrates and p-nitrophenyl esters. Hydrolysis of glycerol ester bonds occurred non-specifically. The purified AML displayed significant stability in the presence of organic solvents (40%, v/v) and catalyzed the synthesis of the flavour ester isoamyl acetate in free and immobilized states.

Original language	English
Pages (from-to)	3373-3379
Number of pages	7
Journal	Bioresource Technology
Volume	102
Issue number	3
DOIs	https://doi.org/10.1016/j.biortech.2010.11.074
Publication status	Published - Feb 2011

Keywords

Actinomycete lipase
Amycolatopsis mediterranei
Characterization
Ester synthesis
Purification

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title = "Purification and properties of Amycolatopsis mediterranei DSM 43304 lipase and its potential in flavour ester synthesis",

abstract = "An extracellular thermostable lipase from Amycolatopsis mediterranei DSM 43304 has been purified to homogeneity using ammonium sulphate precipitation followed by anion exchange chromatography and hydrophobic interaction chromatography. This protocol resulted in a 398-fold purification with 36% final recovery. The purified A. mediterranei DSM 43304 lipase (AML) has an apparent molecular mass of 33 kDa. The N-terminal sequence, AANPYERGPDPTTASIEATR, showed highest similarity to a lipase from Streptomyces exfoliatus. The values of Kmapp and Vmaxapp for p-nitrophenyl palmitate (p-NPP) at the optimal temperature (60. °C) and pH (8.0) were 0.099. ±. 0.010 mM and 2.53. ±. 0.06 mmol/min. mg, respectively. The purified AML displayed significant activity towards a range of short and long chain triglyceride substrates and p-nitrophenyl esters. Hydrolysis of glycerol ester bonds occurred non-specifically. The purified AML displayed significant stability in the presence of organic solvents (40%, v/v) and catalyzed the synthesis of the flavour ester isoamyl acetate in free and immobilized states.",

keywords = "Actinomycete lipase, Amycolatopsis mediterranei, Characterization, Ester synthesis, Purification",

author = "Dheeman, {Dharmendra S.} and Henehan, {Gary T.M.} and Fr{\'i}as, {Jesus M.}",

note = "Funding Information: This work was financially supported by DIT ABBEST Research Scholarship ( PB 03557/2007 ) to DSD. The authors thank Mr. Kevin Conboy, Centre for Synthesis and Chemical Biology (CSCB), University College Dublin (UCD) for technical assistance with gas chromatography.",

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AU - Frías, Jesus M.

N1 - Funding Information: This work was financially supported by DIT ABBEST Research Scholarship ( PB 03557/2007 ) to DSD. The authors thank Mr. Kevin Conboy, Centre for Synthesis and Chemical Biology (CSCB), University College Dublin (UCD) for technical assistance with gas chromatography.

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Purification and properties of Amycolatopsis mediterranei DSM 43304 lipase and its potential in flavour ester synthesis

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Keywords

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