High-yield expression of extracellular lipase from Yarrowia lipolytica and its interactions with lipopeptide biosurfactants: A biophysical approach
Tomasz Janek , Aleksandra Mirończuk , Waldemar Rymowicz , Adam Dobrowolski
AbstractThe unconventional yeast Yarrowia lipolytica is known as a producer of extracellular lipases. Here we overexpressed extracellular lipase (YlLip2) in yeast strain Y. lipolytica AJD ΔXΔA-Lip2 harboring the overexpression cassette of the YALI0A20350 gene under the strong hybrid promoter UAS1B16-TEF. To maintain a high level of YlLip2 production, two extracellular proteases of Y. lipolytica, AEPp and AXPp, were deleted. The purified recombinant YlLip2 presented optimal catalytic activities at 37 °C and pH 8.0. The effect of two lipopeptide biosurfactants, i.e., amphisin produced by Pseudomonas fluorescens DSS73 and viscosinamide secreted by P. fluorescens DR54, on the conformation and activity of YlLip2 was evaluated using spectral methods, surface tension, and the enzyme activity assay. YlLip2 demonstrated high tolerance of the tested biosurfactants and had greater activity retention after incubation with both biosurfactants. Finally, we observed that intrinsic fluorescence intensity of YlLip2 decreased significantly with increasing lipopeptides concentration ranging from 2.5 to 60 μM. Our results showed that both biosurfactants improve enzymatic activity of YlLip2 and might suggest better interaction of the substrate with the active site. These favorable characteristics make YlLip2 a prospective additive in the pharmaceutical, food, cosmetic, and detergent industries.
|Journal series||Archives of Biochemistry and Biophysics, ISSN 0003-9861, e-ISSN 1096-0384, (N/A 100 pkt)|
|Publication size in sheets||0.5|
|Keywords in English||Biosurfactant, Lipopeptide, Yarrowia lipolytica, Lipase, Fluorescence spectroscopy, Surface tension|
|ASJC Classification||; ;|
|License||Journal (articles only); author's original; ; after publication|
|Score||= 100.0, 14-09-2020, ArticleFromJournal|
|Publication indicators||: 2018 = 0.998; : 2018 = 3.559 (2) - 2018=3.269 (5)|
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