Molecular characterization of transesterification activity of novel lipase family I.1

Abstract

Lipase's thermostability and organic solvent tolerance are two crucial properties that enable it to function as a biocatalyst. This study examined the characteristics of two recombinant thermostable lipases (Lk2, Lk3) based on transesterification activity. Conversion of C12-C18 methyl ester with paranitrophenol was investigated in various organic solvent. Both lipases exhibited activity on difference carbon chain length (C12 – C18, C18:1, C18:2) of substrates. The activity of Lk2 was higher in each of substrate compared to that the Lk3. Experimental findings showed that the best substrates for Lk2 and Lk3 are C18:1 and C18:2 respectively, in agreement with the computational analysis. The activity of both enzymes prefers on nonpolar solvent. On nonpolar solvent the enzymes are able to keep its native folding shown by the value of radius gyration, solvent-enzyme interaction and orientation of triad catalytic residues. Lk3 appeared to be more thermostable, with maximum activity at 55°C. The presence of Fe3+ increased the activity of Lk2 and Lk3. However, the activity of both enzymes were dramatically decreased by the present of Ca2+ despite of the enzymes belong to family I.1 lipase known as calcium dependent enzyme. Molecular analysis on His loop of Lk2 and Lk3 on the present of Ca2+ showed that there were shifting on the orientation of catalytic triad residues. All the data suggest that Lk2 and Lk3 are novel lipase on the family I.1 and both lipase available as a biocatalyst candidate.

A thermostable organic solvent-tolerant lipase from Brevibacillus sp.: production and integrated downstream processing using an alcohol-salt-based aqueous two-phase system

“…Due to the advantages of the process, most industrial reactions involving lipases are carried out in non-aqueous media (Ismail et al, 2021 ; Patel and Parikh, 2022 ; Akram et al, 2023 ). Therefore, in addition to stability and activity at elevated temperature and extreme pH values, activity and stability in the presence of organic solvents are essential properties for the industrial application of lipases (Kumar et al, 2016 ; Haryati et al, 2022 ; Ali et al, 2023 ). Lipase SHI-160 was stable in the presence of different polar and non-polar organic solvents.…”

“…However, exposure to organic solvents and/or elevated temperature could pose challenges to enzyme activity and stability (Kumar et al, 2016 ; Kikani et al, 2023 ). Therefore, the use of organic solvent-tolerant thermostable lipases to catalyze reactions in non-aqueous media offers several biotechnological advantages (Ismail et al, 2021 ; Haryati et al, 2022 ; Kikani et al, 2023 ). One approach to develop thermostable enzymes involves isolation and screening of microbes from naturally occurring high-temperature environments, such as hot springs, that harbor a diverse group of microorganisms.…”

Heterologous expression and characterization of a novel thermostable and alkali stable recombinant lipase enzyme from Bacillus thuringensis into E. coliBL21(DE3) for detergent formulation

“…Industrial lipases can be produced from fungi, bacteria, and actinomycetes (Iftikhar et al, 2015). Among these, bacterial lipases showed compatibility at different values of pH and temperature (Haryati et al, 2022). Various bacterial strains, including Bacillus species, have been investigated for lipase production, especially (Hosseini et al, 2023), B. pumilus (Sangeetha et al, 2010b), B. cereus (Wang et al, 2019), B. licheniformis (Sangeetha et al, 2010a), B. subtilis (Ahmed et al, 2010), B. halodurans (Ramchuran et al, 2006), B. coagulans (Mnisi et al, 2005).…”