Expression and Characterization of a Novel Enantioselective Lipase from Aspergillus fumigatus

Abstract

A 1,080-bp cDNA (CGMCC 2873) encoding of a cold-active lipase of Aspergillus fumigatus (AFL67) was cloned and expressed in Escherichia coli for the first time. The new lipase, AFL67, was one-step purified by 8.30 folds through Ni-NTA affinity chromatography with a recovery of 86.8 %. The specific activity of purified AFL67 was 449 U mg(-1) on p-NP hexanoate. AFL67 preferentially hydrolyzed p-nitrophenyl esters of short- and medium-chain fatty acids, with p-nitrophenyl hexanoate the maximum. The optimum temperature and pH was 15 °C and 7.5, respectively. The purified AFL67 was stable at 10-25 °C for 30 min, and in the pH range of 6.0-9.0 for 16 h (at 4 °C). Its activity was increased by 47 and 50 %, in the presence of 10 % (v/v) ethanol and isopropanol, respectively. The new lipase AFL67 highly enantioselectively deacylated (S)-α-acetoxyphenylacetic acid (APA) and o-Cl-APA, m-Cl-APA, and p-Cl-APA to (S)-mandelic acid and its derivates. These features render this cold-active novel lipase AFL67 attractive for biotechnological applications in the field of enantioselective synthesis of chiral mandelic acids, o-acylated mandelic acids, and their derivates and detergent additives.

Cloning, expression, and characterization of Aureobasidium melanogenum lipase in Pichia pastoris

“…XP_750543.1), respectively. Based on BLAST results, Z. brevis lipase showed the second similarity to lipases of Aureobasidium species, and A. fumigatus lipases have been reported with high activity 17,18) ; therefore, the amino acid sequences of lipases among Aureobasidium species and these fungal lipases were aligned for comparison. (Fig.…”

Glyoxalase-based toolbox for the enantioselective synthesis of α-hydroxy carboxylic acids

“…Various purified lipases, 8 dehydrogenases 9 and esterases 10 have been employed for enantioselective synthesis of α-HCAs from α-keto acids, racemic α-hydroxy esters and α-acyloxycarboxylic acids. Such methods that use a single enzyme are quite unattractive because they require the preparation of α-keto acids or esters of α-hydroxy acids to obtain the starting material for enzyme-catalyzed enantioselective reduction or hydrolysis.…”