Lipase from Solvent-Tolerant Pseudomonas sp. DMVR46 Strain Adsorb on Multiwalled Carbon Nanotubes: Application for Enzymatic Biotransformation in Organic Solvents

Appl Biochem Biotechnol (2015) 177:13131326 DOI 10.1007/s12010-015-1816-7

Patel Vrutika1 & Madamwar Datta1

Received: 8 May 2015 /Accepted: 17 August 2015 / Published online: 2 September 2015# Springer Science+Business Media New York 2015

Abstract Immobilization of biocatalysts onto particulate carriers has been widely explored for recycling of biocatalyst. However, surface properties often affect the amount of biocatalysts immobilized, their bioactivity and stability, hampering their wide applications. The aim of this work was to elucidate the importance of nanoimmobilization system in organic synthesis. The surface of multiwalled carbon nanotubes (MWCNTs) was functionalized with a mixture of concentrated acids to create an interface for enzyme immobilization. Successful functionalization and enzyme immobilization was structurally evidenced by transmision electron microscopy analysis and Fourier-transform infrared spectroscopy analysis. Furthermore, immobilized enzyme was exploited for the synthesis of flavoured ester ethyl butyrate in the presence of n-heptane. Optimized conditions for enhanced ester synthesis was found to be8.5 pH, 40 C, 150 rpm, 0.15:0.2 M substrate molar ratio (ethanol/butyric acid) and n-heptane as reaction medium. Utmost 81 % of ester synthesis was obtained using immobilized lipase quite higher in comparison to that of free lipase. The activation energy indicated a lower energy requirement for immobilization of lipase on the surface of functionalized MWCNTs. In summary, immobilization of lipase on functionalized MWCNTs by simple adsorption method displayed excellent properties for enzyme stability and reusability, indicating its potential for application in organic synthesis.

Keywords Multiwalled carbon nanotubes . Ethyl butyrate . Pseudomonas sp . Kinetic study. Biotransformation

* Madamwar Datta [email protected]

Patel Vrutika [email protected]

1 Environmental Genomics and Proteomics Laboratory, BRD School of Biosciences, Sardar Patel

University, Sardar Patel Maidan, Vadtal Road, Satellite Campus, Post Box # 39, Vallabh

Vidhyanagar 388 120 Gujarat, India

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1314 Appl Biochem Biotechnol (2015) 177:13131326

Introduction

Non-aqueous enzymology emerged as a major part of biotechnological research and development area that offers valuable product such as fine chemicals using reactions that are not feasible in aqueous media [1]. Using organic solvents as...