High Pressure Processing of Lipase (Thermomyces lanuginosus): Kinetics and Structure Assessment
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© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, WeinheimLipases are highly demanded enzymes, used to synthesize high commercial value products in food, pharmaceutical, cosmetic, textile, paper, and detergent industries. The effect of high pressure processing (HPP) on Thermomyces lanuginosus lipase is studied at 0.1, 100, and 300 MPa and 25 °C. Kinetics is assessed after HPP treatment in three sets of time series: 0¿25, 51¿75, and 101¿112 min. Conformational change of lipase is studied by infrared attenuated total reflectance (ATR)¿FTIR spectra deconvolution and 1D 1H NMR. Pressure treatment lead to changes in enzymatic activity from 51 to 75 and from 101 to 112 min. Treated enzyme at 300 MPa has the highest activity (¿twofold higher than untreated lipase). Statistical difference in activity (p > 0.05) is not found during the first 25 min. Lipase activity is best described by first-order model (0¿25 min) and zero-order model (51¿75 and 101¿112 min). Lipase unfolding is evident at 100 MPa due to an increase of ß-turns and ß-sheet components. Pressurized enzyme at 300 MPa enhances its activity from 51 to 112 min, due to an increase of ß-sheet and ¿-helix structures (refolding). These findings impact directly on cost-effectiveness of lipase catalyzed products manufacture. Practical Applications: Enzyme enhanced activity and stability improves the performance of manufacture processes. Assessment of structure changes of lipase induced by HPP lead to accurate parameter selection to increase stability and profitability of lipase catalyzed products processing. Pressure levels of 100 MPa led to enzyme unfolding, whereas 300 MPa causes improved activity and stability provoked by lipase refolding. Pressure treatment at 300 MPa of Thermomyces lanuginosus lipase is recommended for long term bioprocessing (¿112 min), which require increased activity and stability of the enzyme.