Characterization of a new sn‐1,3‐regioselective triacylglycerol lipase from Malbranchea cinnamomea

Abstract

The thermophilic ascomycetous fungus Malbranchea cinnamomea produces lipases (EC 3.1.1.3) that allow it to grow efficiently on medium containing triacylglycerol substrates such as plant oils or tributyrin as sole carbon source. In the transcriptome of M. cinnamomea grown on olive oil, we found one cDNA sequence encoding a putative extracellular lipase. This gene, termed as MclipA, was cloned and heterologously expressed in Pichia pastoris. The recombinant protein, rMclipA, catalyzed the hydrolysis of short-chain fatty acid ester such as p-nitrophenyl butyrate (C4) and long-chain fatty acid ester such as p-nitrophenyl myristate (C14). These results indicate that MclipA is a true triacylglycerol lipase. For rMclipA, the optimum lipase activity was obtained at 45 °C, and more than 93% of enzyme activity was retained after 24 H of incubation at temperatures up to 50 °C. rMclipA was active toward p-nitrophenyl esters of various carbon chain lengths with peak activity on long-chain fatty acid (C14). rMclipA displayed high sn-1,3-regioselectivity on hydrolyzing triolein. rMclipA can catalyze oleic acid methyl ester synthesis resulting in a 71% esterification degree after 24 H of reaction at 40 °C. These properties suggest that rMclipA has potential application in, for example, selective hydrolysis of oil, modification of triacylglycerol, and production of biodiesel.

One-Step Partially Purified Lipases (ScLipA and ScLipB) from Schizophyllum commune UTARA1 Obtained via Solid State Fermentation and Their Applications

“…On the other hand, Lotrakul and Dharmsthiti [17] stated that if the lipase is sn -2 regiospecific, 1,3-DAGs and 1(3)-MAG might be produced. According to Tong et al [18], the TLC result obtained with Candida antarctica lipase A (CalA) displayed a clear 1,2- (2,3-) diolein spot and a slightly more intense spot of 1,3-diolein, indicating that CalA was a sn -2 regioselective lipase. For this study, similar scenario can also be observed on the TLC plate in Lanes 4–6 of Figure 4, showing a significant amount of 1,3-DAGs, and less of 1,2- (2,3-) DAGs.…”

Microbial lipases and their industrial applications: a comprehensive review

“…The molecular weight of lipases is in the range of 19–60 kDa and reported to be monomeric protein. The position of the fatty acid in the glycerol backbone, chain length of the fatty acid, and its degree of unsaturation are the factors and the physical properties of lipases depend on it [ 51 , 52 ]. The sensory and nutritive values of given triglyceride also affected by these features.…”

An overview on the conversion of glycerol to value‐added industrial products via chemical and biochemical routes

“…Lipases are a particular class of enzymes with a vast industrial application range and that can be used in the synthesis of glycerol-derived products [21]. They present great versatility, being used in a range of industrial reactions, such as asymmetric esterification and transesterification, aminolysis, asymmetric hydrolysis, among others [3,13,19,[22][23][24][25][26][27][28][29][30][31][32][33][34].…”