Heterologous expression and characterization of a new lipase from Pseudomonas fluorescens Pf0–1 and used for biodiesel production

Abstract

As a kind of important biocatalysts, Pseudomonas lipases are commonly applied in various industrial fields. Pflip1, a new extracellular lipase gene from Pseudomonas. fluorescens Pf0–1, was first cloned and respectively expressed in Escherichia coli BL21(DE3) and Pichia pastoris KM71, the recombinant proteins Pflip1a and Pflip1b were later purified separately. Then Pflip1a was further characterized. The optimum pH of Pflip1a was 8.0 and its optimal temperature was 70 °C. After incubation at 70 °C for 12 h, Pflip1a could retain over 95% of its original activity. It showed the highest activity towards p-nitrophenyl caprylate. Moreover, its activity was profoundly affected by metal ion, ionic surfactants and organic solvents. Furthermore, the two obtained recombinant lipases were immobilized on the magnetic nanoparticles for biodiesel preparation. The GC analysis showed that for the immobilized lipases Pflip1b and Pflip1a, the biodiesel yield within 24 h respectively attained 68.5% and 80.5% at 70 °C. The activities of the two immobilized lipases still remained 70% and 82% after 10 cycles of operations in non-solvent system. These characteristics and transesterification capacity of the recombinant protein indicated its great potential for organic synthesis, especially for biodiesel production.

Purification, Characterization and Immobilization of Lipase from Proteus vulgaris OR34 for Synthesis of Methyl Oleate

“…It has been reported that calcium ions are needed for lipolytic activity of some bacterial lipases. In fact, Liu et al [50] demonstrated that lipase from Pseudomonas fluorescens Pf0-1, which belongs to sub-family I.1, was stimulated (160%) in the presence of 10 mM CaCl 2 . Moreover, the activity of lipase from Staphylococcus hyicus (sub-familyI.6) increased about 40-fold in the presence of Ca 2+ ions and was strongly inhibited in the presence of chelators [51].…”

The occurrence, enzymatic production, and application of ethyl butanoate, an important flavor constituent

“…29,56,57 But this reaction is extensively used for biodiesel production from natural oils. [58][59][60][61][62] Acidolysis is another type of transesterification reaction where esters are produced by the reaction of an ester and an acid.…”

“…Alcoholysis reaction for the production of ethyl butanoate is rare in use, mostly happens in lactic acid bacteria (LAB) in fermented dairy products 29,56,57 . But this reaction is extensively used for biodiesel production from natural oils 58‐62 …”

Production of thermostable multiple enzymes from Bacillus amyloliquefaciens KUB29

“…Atividade lipolítica aumentada com a adição de Ca 2+ e Ba 2+ e reduzida na presença de Cu 2+ , Zn 2+ e Fe 2+ foi reportada para P. fluorescens Pf0-1 (Liu W, Li M, Yan Y, 2017).…”