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Purification and Characterization of a Lipase with High Thermostability and Polar Organic Solvent-Tolerance from Aspergillus niger AN0512

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Lipids

Abstract

An extracellular lipase (EC 3.1.1.3, AN0512Lip) from Aspergillus niger AN0512 was purified and its characteristics were investigated. After the process of ammonium sulfate precipitation followed by ion-exchange chromatography and gel filtration, the purified lipase was achieved with 203.6-fold purification and 22.1 % recovery. AN0512Lip exhibited the highest activity at 50 °C and pH 5.0. It was thermostable and pH-stable, as indicated by that more than 50 % activity retained at 60 °C for 20 h and more than 90 % activity retained at pH 3.0 for 20 h, respectively. AN0512Lip activity was stimulated by some divalent metal ions (especially Cu2+, Ca2+), while greatly suppressed by EDTA, indicating that AN0512Lip was a metal-activated enzyme. Moreover, AN0512Lip exhibited high tolerance for various polar organic solvents with log P < 0.8, and the highest lipase activity (476 % of its original activity) was achieved after addition of 90 % (V/V) isopropanol to the reaction mixture. AN0512Lip also displayed 3-regiospecificity and great affinity for the long-chain fatty ester. The preliminary test showed that AN0512Lip was a candidate for enriching EPA and DHA in fish oil. All the unique properties, such as thermostability, Cu2+-dependent, 3-regiospecificity, and polar organic solvent-tolerance, indicated that AN0512Lip could have potential applications in the food industry, even in organic synthesis and the pharmaceutical industry.

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Abbreviations

AN0512Lip:

Lipase from Aspergillus niger AN0512

DH:

Degree of hydrolysis

EDTA:

Ethylene diamine tetraacetic acid

K m :

Michaelis constant

PMSF:

Phenylmethanesulfonyl fluoride

p-NPP:

p-Nitrophenyl palmitate

p-NPD:

p-Nitrophenyl dodecanoate

p-NPO:

p-Nitrophenyl octanoate

p-NPA:

p-Nitrophenyl acetate

SDS:

Sodium dodecyl sulfonate

TLC:

Thin-layer chromatography

V max :

The maximum velocity for the reaction

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Acknowledgments

We are grateful for financial support from the National Natural Science Fund of China (Grant Numbers 31130042, 31171630 and 21206046) and the Specialized Research Fund for the Doctoral Program of Higher Education of China (Number 20130172110018).

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Authors and Affiliations

  1. School of Light Industry and Food Sciences, South China University of Technology, Guangzhou, 510641, Guangdong, China

    Guang Liu, Songqing Hu, Lin Li & Yi Hou

  2. Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, South China University of Technology, Guangzhou, 510641, Guangdong, China

    Songqing Hu & Lin Li

  3. State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou, 510641, Guangdong, China

    Yi Hou

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  1. Guang Liu
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Correspondence to Yi Hou.

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Liu, G., Hu, S., Li, L. et al. Purification and Characterization of a Lipase with High Thermostability and Polar Organic Solvent-Tolerance from Aspergillus niger AN0512. Lipids 50, 1155–1163 (2015). https://doi.org/10.1007/s11745-015-4052-6

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  • DOI: https://doi.org/10.1007/s11745-015-4052-6

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