Activation and deformation of immobilized lipase on self-assembled monolayers with tailored wettability

Abstract

In this work, lipase from Candida rugosa (CRL) was immobilized on self-assembled monolayers (SAMs) with various wettabilities ranging from highly hydrophilic to highly hydrophobic by adsorption in order to clearly elucidate the interfacial activation character of lipases. The SAMs were made of 11-hydroxyundecane-1-thiol and 1-dodecanethiol. The adsorption behavior was monitored in situ by quartz crystal microbalance with dissipation (QCM-D), and the enzyme binding constants indicated a stronger affinity between CRL and more hydrophobic surfaces. Atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) were used to characterize the morphologies of the adsorbed lipases. Amide I band attenuated total reflection/Fourier transformed infrared (ART/FTIR) spectroscopy showed an increasing fraction of intermolecular β-sheet content on surfaces with higher hydrophilicities. Moreover, liquid chromatography (LC) verified that the activity of CRL adsorbed on a hydrophobic surface was higher than that of CRL adsorbed on a hydrophilic surface. This work related the enzyme activity to the substrate properties, adsorption behavior, distribution, and morphology of lipases, helping to achieve the external control of both the immobilization process and enzyme utilization.

Hydrophobic pore space constituted in macroporous ZIF-8 for lipase immobilization greatly improving lipase catalytic performance in biodiesel preparation

“…Therefore, the amide I band was used herein. A second derivative analysis to locate peaks due to secondary structural components was adopted [24,29]. After second derivative analysis of amide I (1600-1700 cm −1 ) bands, the second derivatives of the spectra are shown in Fig.…”

“…Considering the hydrophilic property of glycerol may result in the adsorption on the lipase, hydrophobic modification on the carrier of the immobilized lipase may lessen the adsorption of glycerol to a great extent. Despite of the fact that some researches observed the performance improvement of immobilized lipase by enhancing the hydrophobicity of supports mainly through influencing enzyme conformation [24][25][26], the influence on glycerol adsorption during biodiesel production has not been reported.…”

Esterification of oleic acid with methanol by immobilized lipase on wrinkled silica nanoparticles with highly ordered, radially oriented mesochannels

“…They achieved a fatty acid methyl ester (FAME) yield of 95.2% with 5:1 M ratio of methanol to oil and 4% Novozym 435 at 45°C for 10 h. The type of support influences the activity and operational stability of immobilized lipases. Numerous supports have been investigated, including macroporous and microporous polymers, silica sol-gel matrix or aerogels, ordered mesoporous silica, and other porous ceramics [13][14][15][16]. In recent years, mesoporous silica nanoparticles have received increasing attention in enzyme and protein immobilization because of their tunable and uniform pore system, high specific surface area, great compatibility, and low toxicity.…”

Adsorption and Activity of Lipase on Polyphosphazene-Modified Polypropylene Membrane Surface

“…Poly(n-butylamino) 1.67 (allylamino) 0.33 phosphazene film had a water contact angle of 113.6 • , meaning a 30 • increase compared with that of poly(allylamino)phosphazene. In the previous work done by us, lipase was found to adsorb on the support easily with a water contact angle of 105.9 • , with a higher enzyme binding constant value, and the hydrophobic interaction can expose the catalytic site and produce even and regular protein deposition, which activate the lipase [16]. Thus we used poly(n-butylamino) 1 …”

“…Therefore, special emphasis has been put on immobilizing lipases onto hydrophobic support surfaces in recent years, which is based on the assumption that the active state of lipases can be stabilized by the hydrophobic interaction between the hydrophobic active center of lipases and the hydrophobic support [12][13][14][15]. In a previous study, we systematically studied the structural rearrangements, immobilization kinetics and protein aggregation of lipase from Candida rugosa absorbed on a wettability-tailored surface, and concluded that a support with proper hydrophobicity is of vital importance for obtaining a highly efficient immobilized lipase [16].…”