Effect of intense pulsed light on the deactivation of lipase: Enzyme-deactivation kinetics and tertiary structural changes by fragmentation.

Abstract:

:The effect of intense pulsed light (IPL) irradiation on Chromobacterium viscosum lipase was investigated with a primary focus on catalytic activity and molecular structure. During IPL irradiation, lipase activity decreased significantly with increasing pulse fluence (Fp) and exposure time (te). IPL-induced deactivation kinetics were further elucidated based on a two-step series-type deactivation model (constant deactivation rate k1 >k2). Fp was found to be the dominant variable affecting the degree of lipase deactivation, and residual activity was not associated with increasing te below a certain Fp energy density (2.66 mJ/cm2), implying a critical threshold for IPL-induced deactivation of lipase. From the results of fluorescence spectroscopy and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), we determined that IPL-induced deactivation was caused by fragmentation, leading to lipase tertiary structural changes. Furthermore, the results of FindPept analysis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) indicated that the internal sensitive bonds of lipase were cleaved preferentially by IPL, such that IPL irradiation induced site-sensitive fragmentation and peptide bond cleavage.

journal_name

Enzyme Microb Technol

authors

Jeon MS,Park KM,Yu H,Park JY,Chang PS

doi

10.1016/j.enzmictec.2019.02.001

subject

Has Abstract

pub_date

2019-05-01 00:00:00

pages

63-69

eissn

0141-0229

issn

1879-0909

pii

S0141-0229(18)30185-6

journal_volume

124

pub_type

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