An active-site titration method for lipases

• A method for active-site titration of lipases has been developed based on irreversible inhibition by methyl p-nitrophenyl n-hexylphosphonate. This method was applied to five lipases displaying from minor to pronounced interfacial activation. Soluble and immobilized lipases were successfully titrated in aqueous media. A low concentration of sodium dodecyl sulfate was needed for lipases displaying pronounced interfacial activation. The carrier of some of the immobilized preparations adsorbed part of the produced p-nitrophenolate, This problem could be solved by extracting the p-nitrophenolate after inhibition. The method was extended to apolar organic solvents in the case of immobilized lipase preparations.

Nyckelord

concentration

immobilized lipase

aqueous medium

organic medium

interfacial activation

detergent

inhibitor

serine proteases

interfacial activation

organic-chemistry

in-vitro

biocatalysts

inhibition

solvents

cutinase

titrant

enzymes

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)