A comparative study on two fungal lipases from Thermomyces lanuginosus and Yarrowia lipolytica shows the combined effects of detergents and pH on lipase adsorption and activity.
The effects of various detergents and pH on the interfacial binding and activity of two fungal lipases from Yarrowia lipolytica (YLLIP2) and Thermomyces lanuginosus (TLL) were investigated using trioctanoin emulsions as well as monomolecular films spread at the air-water interface. Contrary to TLL, YLLIP2 was found to be more sensitive than TLL to interfacial denaturation but it was protected by detergent monomers and lowering the temperature. At pH 7.0, both the interfacial binding and the activities on trioctanoin of YLLIP2 and TLL were inhibited by sodium taurodeoxycholate (NaTDC). At pH 6.0, however, YLLIP2 remained active on trioctanoin in the presence of NaTDC, whereas TLL did not. YLLIP2 activity on trioctanoin was associated with strong interfacial binding of the enzyme to trioctanoin emulsion, whereas TLL was mostly detected in the water phase. The combined effects of bile salts and pH on lipase activity were therefore enzyme-dependent. YLLIP2 binds more strongly than TLL at oil-water interfaces at low pH when detergents are present. These findings are particularly important for lipase applications, in particular for enzyme replacement therapy in patients with pancreatic enzyme insufficiency since high detergent concentrations and highly variable pH values can be encountered in the GI tract.
Aloulou
Ahmed
A
Laboratory of Enzymology at Interfaces and Physiology of Lipolysis, CNRS-UPR 9025-IBSM, 31 Chemin Joseph-Aiguier, 13009 Marseille, France.
Puccinelli
Delphine
D
De Caro
Alain
A
Leblond
Yves
Y
Carrière
Frédéric
F
D000327:Adsorption; D000595:Amino Acid Sequence; D000818:Animals; D002210:Caprylates / Q000737:chemistry; D003902:Detergents / Q000737:chemistry*; D004795:Enzyme Stability; D005079:Excipients / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016415:Sequence Alignment; D013499:Surface Properties; D013696:Temperature; D014280:Triglycerides / Q000737:chemistry; D025062:Yarrowia / Q000201:enzymology*
D002210:Caprylates; D003902:Detergents; D005079:Excipients; D005656:Fungal Proteins; D014280:Triglycerides; C003637:tricaprylin; D008049:Lipase