Isolation and characterization of a novel thermo-solvent-stable lipase from Pseudomonas brenneri and its application in biodiesel synthesis

Priyanka Priyanka,Barry J. Ryan,Gary T. Henehan,Gemma K. Kinsella

doi:10.1016/j.bcab.2020.101806

Priyanka Priyanka, Barry J. Ryan + Show 2 more

Open Access

https://doi.org/10.1016/j.bcab.2020.101806

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Abstract

Abstract Pseudomonads are one of the most studied species of bacteria as they display remarkable metabolic and physiological versatility. This enables them to colonize a wide variety of terrestrial and aquatic habitats, generating biotechnologically interesting enzymes. Here, the partial purification and characterization of a novel, extracellularly-produced, lipase from Pseudomonas brenneri is described. The partially purified lipase was active over a broad pH range (5.0–9.0) and was stable at 70 °C for 45 min. The lipase displayed significant stability, and in some cases activation, in the presence of organic solvents with log P ≥ 2.0. Such stability characteristics indicated that this lipase could potentially be useful as a biocatalyst for biodiesel production. This was subsequently demonstrated through the facile production of Fatty Acid Methyl Esters in the presence of olive oil and methanol. Possible applications for this novel, stable lipase include the bioremediation of oil in the environment.

Highlights

Enzyme biotechnology has progressed from catalysis in an aqueous medium to synthesis in nonaqueous media
A lipase from P. cepacia has been used for the synthesis of cinnamyl propionate (Badgujar, Pai and Bhanage, 2016) while lipases from P. fluorescens and P. stutzeri have been used in the cosmetic, oral care and pharmaceutical industries (Vescovi et al, 2017; Cao et al, 2012)
Isolation and identification of solvent tolerant lipase producing strain Two lipolytic cultures isolated from soil samples (GPS location 53°00'12.4"N, 6°20'47.9"W; a woodland in a national park) were stable in cyclohexane, ethanol, n-hexane, heptane and methanol by plate overlay method (Patel, Nambiar and Madamwar, 2014). 16S rRNA sequencing of these strains identified one of the lipolytic cultures as Pseudomonas reinekei while the other isolate was identified as Pseudomonas brenneri

Summary

Introduction

Enzyme biotechnology has progressed from catalysis in an aqueous medium to synthesis in nonaqueous media. Solvent stable lipases are key biocatalysts in non-aqueous solutions and can catalyse various reactions (such as esterification, alcoholysis, acidolysis and interesterification; (Sharma and Kanwar, 2014)). Due to their wide ranging applications, lipases have long been subjected to intensive study. Based on initial characterisation and this application, the novel lipase will be of further interest for various biocatalytic and biotechnological uses in non-aqueous media, as well as bioremediation of oil in the environment (Kumar, 2020)

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