Characterization of a novel lipase from Pseudomonas aeruginosa

Nodir Sh Berdiev,Jamolitdin F Ziyavitdinov,Shukhratjon S Olimjonov,Shavkat I Salikhov,Akmal M Asrorov

doi:10.2478/nbec-2019-0006

Abstract

Abstract Lipases cleaving oils into fatty acids and glycerol are of great interest for the use in increasing the efficiency of fuels. In this work, a novel lipase from Pseudomonas aeruginosa, P. aeruginosa A12, was isolated by ion-exchange and hydrophobic chromatography. The purity of lipase was shown by electrophoresis and its molecular weight was estimated to be ~ 31.6 kDa. The whole amino acid sequence was analyzed by an LC-MS/MS method. Temperature- and pH-dependent optimum of the enzyme compiled 30 °C and 7.5, respectively. The obtained enzyme exhibited 79 % similarity of amino acid sequence to a lipase isolated from the same strain of P. aeruginosa. Thus, the novel lipase was determined to belong to I.1 subfamily of bacterial true lipases. Three dimensional structure of the isolated lipase isoform was modeled based on obtained sequences. Amino acids forming the catalytic domain were shown in the model. Lid domain is suggested to be in the open conformation. These results provide a potential alternative for enzymatic digestion of fuel oils and serve for the development of fundamental knowledge of lipase activity.

Highlights

Lipases (EC 3.1.1.3) are triglyceride ester hydrolases that cleave ester bonds in glycerides (Balcao et al 1996)
The growth dynamics of cells and kinetic curves of lipase activity in bacterial cultures were determined during 4 d of cultivation period in 12 h intervals
The data obtained using 1 % (v/v) cottonseed oil as inductor of lipases show that the lipase activity in cultured bacteria C. violaceum reached the maximum after 48 h and 84 h were determined as the maximum point for

Summary

Introduction

Lipases (EC 3.1.1.3) are triglyceride ester hydrolases that cleave ester bonds in glycerides (Balcao et al 1996). The enzymes isolated from P. aeruginosa reveal different MW, pH and temperature optimum. A 54 kDa lipase isolated from P. aeruginosa was determined to have pH optimum in the range 6 – 9 (Karadzic et al 2006).

Results

Conclusion