Real-time assessment and characterization of immobilized lipase onto a natural matrix and qualitative reaction kinetic s
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ORIGINAL ARTICLE
Real‑time assessment and characterization of immobilized lipase onto a natural matrix and qualitative reaction kinetic studies using swept‑source optical coherence tomography Muktesh Mohan1 · Zoya Alam1 · Raju Poddar1 · Soham Chattopadhyay1 Received: 9 May 2020 / Accepted: 21 August 2020 © King Abdulaziz City for Science and Technology 2020
Abstract The current study presents a method based on Optical Coherence Tomography (OCT) for non-destructive, real-time analysis and portrayal of immobilization efficacy for lipase on a natural matrix namely, eggshell. Subsequently, qualitative biochemical reaction kinetics of immobilized lipase was also studied. Successful immobilization of lipase on eggshell was confirmed by the presence of a clear peak in ‘A’ scan of OCT image. From immobilization kinetics it is clearly observed that the thickness of the highest peak of the A-scan increases significantly and peak intensity saturated after 90 min of incubation. Hydrolysis of oil using immobilized lipase indicated that the release of free fatty acids increased up to 8 h during reaction and the result was in accordance with the ‘B’ scan data of the OCT system. Changes in scattering coefficient-based analysis were performed with respect to incubation time to showcase the immobilization process and hydrolysis reaction of lipase. Scanning electron microscope analysis with smoother surface indicated presence of lipase on eggshell matrices, with no further change after oil hydrolysis. Keywords Lipase · Eggshell · Enzyme-immobilization · Swept-source optical coherence tomography (SSOCT) · Reaction kinetics · Scattering coefficients
Introduction Triacylglycerol lipase (EC: 3.1.1.3) is one of the most important enzymes that catalyses oil into free fatty acid (FFA) and glycerol (Ribeiro et al. 2011). Industrial-scale production of lipase is primarily restricted to the sources such as bacteria, fungi or animal pancreas (Zhang et al. 2012; Yang et al. 2009; Kumari et al. 2009); whereas plant-based lipase (primarily from oilseeds, rice, wheat, maize etc.) are gaining importance nowadays because of their availability and higher yield (Seth et al. 2014). Lipases are mostly used in biodiesel production, pharmacy industry, cosmetics manufacturing, paper and pulp industry etc. (Paul et al. 2015; Chattopadhyay et al. 2011; Jaeger et al. 1999). * Raju Poddar [email protected] * Soham Chattopadhyay [email protected] 1
Department of Bio‑engineering, Birla Institute of Technology-Mesra, Ranchi 835215, India
Several researchers reported that using immobilized lipase is the better alternative compared to free one due to its enhanced stability, reusability and cost-effectiveness (Yilmaz et al. 2011; Hernandez et al. 2011). Lipase immobilized on eggshell reported to be superior compared to other matrices, and its reusability, stability, and specificity were proved to be a better alternative than other matrices. It was already established that the immobilization of lipase on eggshell is purely an adsorption process without
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