Biochemical and structural characterization of triacylglycerol lipase from Penicillium cyclopium

Abstract

An extracellular lipase, active on water-insoluble triacylglycerols, has been isolated from Penicillium cyclopium. The purified enzyme has a molecular mass of 29 kDa by gel filtration and SDS-polyacrylamide gel electrophoresis. It hydrolyzes emulsions of tributyrin, trioctanoin, and olive oil at the same rate as pancreatic lipase and shows very low activity against partial acylglycerols (monooctanoin and dioctanoin) and methyl esters. It is stable at 35 degrees C for 60 min and has maximal activity in a pH range of 8-10. Hydrolysis of triacylglycerols by P. cyclopium lipase is inhibited by detergents such as Triton X-100. Comparison of the sequence of the 20 first amino acid residues of P. cyclopium triacylglycerol lipase with other Penicillium lipases indicates a high homology with previously characterized lipases produced by P. expansum and P. solitum which are enzymes of comparable size and substrate specificity. Conversely, homology between P. cyclopium lipase and P. simplicissimum lipase, a nonspecific lipolytic enzyme, is low. Penicillium cyclopium triacylglycerol lipase shows no homology with P. camembertii lipase which is specific to monoacylglycerol and diacylglycerol.

Biochemistry of lipolytic enzymes secreted byPenicillium solitumandCladosporium cladosporioides

“…The value obtained for P. solitum 194A was in line with the most well-known lipases from the genus Penicillium (a molecular mass in the range of 25-43 kDa), 29,34) similar to the value obtained for the type II lipases from P. camembertii (38 kDa) 5,35,36) and P. cyclopium (40-43 kDa). 5,35) Conversely, the value differed from the molecular mass of the type I P. solitum lipase previously reported (27 kDa). 5,35,37) This difference is likely to be due to an isoform of the enzyme secreted by this micro-organism.…”

“…5,35) Conversely, the value differed from the molecular mass of the type I P. solitum lipase previously reported (27 kDa). 5,35,37) This difference is likely to be due to an isoform of the enzyme secreted by this micro-organism. Evidence of an additional lipase secreted by P. solitum is noteworthy, because it supports the suggestion that penicillia are capable of producing more than one extracellular lipase.…”

Potencial de biocatálise enantiosseletiva de lipases microbianas

“…Em relação as cepas de Penicillium, vários trabalhos foram encontrados envolvendo produção, caracterização e purificação de lipases de diferentes espécies, como P. citrinum 36 , P. cyclopium 37 , P. simplicissimum 38 , P. caseicolum 39 , P restrictum 40 , P. expansum 41 , P. chrysogenum 42 , P. roqueforti 43 , P. camembertii 44 , P. abeanum 45 , entre outras espécies 46 . Entretanto, parece que este é o primeiro relato sobre a produção de lipase pelo fungo Penicillium solitum.…”

Production of extracellular lipases by Rhizopus oligosporus in a stirred fermentor

“…Different microorganisms have been used for the production of lipases such as Rhizopus species, Rhizomucor mehei, Aspergillus niger, Penicillium species, Bacillus subtilus (12,13,14,25). Gopinath et al (9) reported about 34 wild fungal species associated with edible oil mill wastes which were isolated by the serial dilution technique.…”