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Purification and Characterization of a Lipase with High Thermostability and Polar Organic Solvent‐Tolerance from Aspergillus niger AN0512
Author(s) -
Liu Guang,
Hu Songqing,
Li Lin,
Hou Yi
Publication year - 2015
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-015-4052-6
Subject(s) - thermostability , chemistry , lipase , ammonium sulfate precipitation , aspergillus niger , solvent , size exclusion chromatography , chromatography , nuclear chemistry , organic chemistry , enzyme , biochemistry
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 Cu 2+ , Ca 2+ ), 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, Cu 2+ ‐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.