Overexpression of <i>Fusarium solani</i> lipase in <i>Pichia pastoris</i> and its application in lipid degradation

Wongwatanapaiboon, Jinaporn; Malilas, Waraporn; Ruangchainikom, Chalermchai; Thummadetsak, Gamgarn; Chulalaksananukul, Suphang; Marty, Alain; Chulalaksananukul, Warawut

doi:10.1080/13102818.2016.1202779

Cited by 16 publications

(7 citation statements)

References 32 publications

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“…The developed method was also successfully applied to unconcentrated crude microbial culture media. We were able to detect lipolytic enzymes produced by Aspergillus aculeatus [ 29 ], Colletotrichum gloeosporioides [ 30 , 31 , 32 ], Fusarium solani [ 33 , 34 , 35 , 36 , 37 , 38 ], Penicillium expansum [ 6 , 39 , 40 , 41 , 42 ], and Trichoderma harzianum [ 43 , 44 , 45 , 46 , 47 ]. The choice of substrates used in chromogenic agar detection is not limited to triglycerides, but could also extended to di- or monoglycerides, vinyl laurate and other acyl ester substrates, which would be beneficial for screening of partial-glyceride lipases or steryl esterases.…”

Section: Discussionmentioning

confidence: 99%

Zymography for Picogram Detection of Lipase and Esterase Activities

Zhang

Nguyen

2021

Molecules

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Lipases and esterases are important catalysts with wide varieties of industrial applications. Although many methods have been established for detecting their activities, a simple and sensitive approach for picogram detection of lipolytic enzyme quantity is still highly desirable. Here we report a lipase detection assay which is 1000-fold more sensitive than previously reported methods. Our assay enables the detection of as low as 5 pg and 180 pg of lipolytic activity by direct spotting and zymography, respectively. Furthermore, we demonstrated that the detection sensitivity was adjustable by varying the buffering capacity, which allows for screening of both high and low abundance lipolytic enzymes. Coupled with liquid chromatography-mass spectrometry, our method provides a useful tool for sensitive detection and identification of lipolytic enzymes.

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Section: Discussionmentioning

confidence: 99%

Zymography for Picogram Detection of Lipase and Esterase Activities

Zhang

Nguyen

2021

Molecules

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“…Since it is a strong constitutive promoter, no induction step is required, resulting in simpler and shorter fermentation cultivations. The expression of Fusarium solani NAN103 lipase (FSL) under pGAP or pAOX1 showed that pGAP-FSL produced the highest specific lipase activity and, in addition, there was a two-day reduction in the cultivation time [88]. Many promoters with different strengths and properties are available for expression in K. phaffii.…”

Section: Eukaryotic Expression Systemsmentioning

confidence: 99%

Advances in Recombinant Lipases: Production, Engineering, Immobilization and Application in the Pharmaceutical Industry

et al. 2020

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Lipases are one of the most used enzymes in the pharmaceutical industry due to their efficiency in organic syntheses, mainly in the production of enantiopure drugs. From an industrial viewpoint, the selection of an efficient expression system and host for recombinant lipase production is highly important. The most used hosts are Escherichia coli and Komagataella phaffii (previously known as Pichia pastoris) and less often reported Bacillus and Aspergillus strains. The use of efficient expression systems to overproduce homologous or heterologous lipases often require the use of strong promoters and the co-expression of chaperones. Protein engineering techniques, including rational design and directed evolution, are the most reported strategies for improving lipase characteristics. Additionally, lipases can be immobilized in different supports that enable improved properties and enzyme reuse. Here, we review approaches for strain and protein engineering, immobilization and the application of lipases in the pharmaceutical industry.

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“…The accumulation of lipid in the cells occurs via a dynamic equilibrium of fatty acid synthesis and degradation [31]. When degraded by lipases, TAGs are hydrolyzed into free fatty acids and glycerol [32]. Production of glycerol during the course of fungal cultivation in CWFS has probably been performed though a similar mechanism.…”

Section: Production Of Fungal Biomass For Incorporation Into Pectin Fmentioning

confidence: 99%

A Solvent-Free Approach for Production of Films from Pectin and Fungal Biomass

et al. 2018

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Self-binding ability of the pectin molecules was used to produce pectin films using the compression molding technique, as an alternative method to the high energy-demanding and solvent-using casting technique. Moreover, incorporation of fungal biomass and its effects on the properties of the films was studied. Pectin powder plasticized with 30% glycerol was subjected to heat compression molding (120 °C, 1.33 MPa, 10 min) yielding pectin films with tensile strength and elongation at break of 15.7 MPa and 5.5%, respectively. The filamentous fungus Rhizopus oryzae was cultivated using the water-soluble nutrients obtained from citrus waste and yielded a biomass containing 31% proteins and 20% lipids. Comparatively, the same strain was cultivated in a semi-synthetic medium resulting in a biomass with higher protein (60%) and lower lipid content (10%). SEM images showed addition of biomass yielded films with less debris compared to the pectin films. Incorporation of the low protein content biomass up to 15% did not significantly reduce the mechanical strength of the pectin films. In contrast, addition of protein-rich biomass (up to 20%) enhanced the tensile strength of the films (16.1-19.3 MPa). Lastly, the fungal biomass reduced the water vapor permeability of the pectin films.

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Overexpression of Fusarium solani lipase in Pichia pastoris and its application in lipid degradation

Cited by 16 publications

References 32 publications

Zymography for Picogram Detection of Lipase and Esterase Activities

Zymography for Picogram Detection of Lipase and Esterase Activities

Advances in Recombinant Lipases: Production, Engineering, Immobilization and Application in the Pharmaceutical Industry

A Solvent-Free Approach for Production of Films from Pectin and Fungal Biomass

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