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Lodens S, Roelants SLKW, Ciesielska K, Geys R, Derynck E, Maes K, Pattyn F, Van Renterghem L, Mottet L, Dierickx S, Vanhaecke L, Devreese B, De Maeseneire SL, Soetaert W
Unraveling and resolving inefficient glucolipid biosurfactants production through quantitative multiomics analyses of Starmerella bombicola strains
Biotechnology and Bioengineering 117(2) (2020)
453-465
Starmerella bombicola
(NCBI TaxID 75736,
species name lookup)
Taxonomic group: fungi / Ascomycota
(Phylum: Ascomycota)
NCBI PubMed ID: 31612987Publication DOI: 10.1002/bit.27191Journal NLM ID: 7502021Publisher: New York: Wiley-VCH
Correspondence: Sophie.Roelants
ugent.be
Institutions: Centre for Industrial Biotechnology and Biocatalysis (InBio.be), Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium, Bio Base Europe Pilot Plant, Desteldonk, Belgium, L-Probe, Department of Sciences, Ghent University, Ghent, Belgium, Center for Medical Genetics Ghent, Ghent University, Ghent, Belgium, Flamac, Zwijnaarde, Belgium, Laboratory of Chemical Analysis, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium
Glucolipids (GLs) are glycolipid biosurfactants with promising properties. These GLs are composed of glucose attached to a hydroxy fatty acid through a ω and/or ω-1 glycosidic linkage. Up until today these interesting molecules could only be produced using an engineered Starmerella bombicola strain (∆ugtB1::URA3 G9) producing GLs instead of sophorolipids, albeit with a very low average productivity (0.01 g/L/h). In this study, we investigated the reason(s) for this via reverse-transcription quantitative polymerase chain reaction and Liquid chromatography-multireaction monitoring-mass spectrometry. We found that all glycolipid biosynthetic genes and enzymes were downregulated in the ∆ugtB1 G9 strain in comparison to the wild type. The underlying reason for this downregulation was further investigated by performing quantitative metabolome comparison of the ∆ugtB1 G9 strain with the wild type and two other engineered strains also tinkered in their glycolipid biosynthetic gene cluster. This analysis revealed a clear distortion of the entire metabolism of the ∆ugtB1 G9 strain compared to all the other strains. Because the parental strain of the former was a spontaneous ∆ura3 mutant potentially containing other "hidden" mutations, a new GL production strain was generated based on a rationally engineered ∆ura3 mutant (PT36). Indeed, a 50-fold GL productivity increase (0.51 g/L/h) was obtained with the new ∆ugtB1::URA3 PT36 strain compared with the G9-based strain (0.01 g/L/h) in a 10 L bioreactor experiment, yielding 118 g/L GLs instead of 8.39 g/L. Purification was investigated and basic properties of the purified GLs were determined. This study forms the base for further development and optimization of S. bombicola as a production platform strain for (new) biochemicals.
fermentation, purification, glucolipid, Starmerella bombicola, strain engineering, (bio)surfactants
Structure type: cyclic polymer repeating unit ; n=1
Location inside paper: Fig. 1, diacetylated lactonic SL
Compound class: glycolipid, sophorolipid
Contained glycoepitopes: IEDB_140628,IEDB_142488,IEDB_146664,IEDB_983931,SB_192
Methods: ESI-MS, enzymatic digestion, extraction, LC-MS, cell growth, LC-MS/MS, precipitation, sonication, antimicrobial assay, centrifugation, qRT-PCR, optical density measurement, UPLC-ELSD, RPLC
Enzymes that release or process the structure: cyp52m1 (monooxygenase), ugta1 (glucosyltransferase), ugtb1 (glucosyltransferase), At (acetyltransferase), sble
Biosynthesis and genetic data: biochemical data, genetic data
Related record ID(s): 50967
NCBI Taxonomy refs (TaxIDs): 75736
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There is only one chemically distinct structure:
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Lodens S, Roelants SLKW, Ciesielska K, Geys R, Derynck E, Maes K, Pattyn F, Van Renterghem L, Mottet L, Dierickx S, Vanhaecke L, Devreese B, De Maeseneire SL, Soetaert W
Unraveling and resolving inefficient glucolipid biosurfactants production through quantitative multiomics analyses of Starmerella bombicola strains
Biotechnology and Bioengineering 117(2) (2020)
453-465
Starmerella bombicola (ΔugtB1::ura3 G9 mutant)
(Ancestor NCBI TaxID 75736,
species name lookup)
Taxonomic group: fungi / Ascomycota
(Phylum: Ascomycota)
NCBI PubMed ID: 31612987Publication DOI: 10.1002/bit.27191Journal NLM ID: 7502021Publisher: New York: Wiley-VCH
Correspondence: Sophie.Roelants
ugent.be
Institutions: Centre for Industrial Biotechnology and Biocatalysis (InBio.be), Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium, Bio Base Europe Pilot Plant, Desteldonk, Belgium, L-Probe, Department of Sciences, Ghent University, Ghent, Belgium, Center for Medical Genetics Ghent, Ghent University, Ghent, Belgium, Flamac, Zwijnaarde, Belgium, Laboratory of Chemical Analysis, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium
Glucolipids (GLs) are glycolipid biosurfactants with promising properties. These GLs are composed of glucose attached to a hydroxy fatty acid through a ω and/or ω-1 glycosidic linkage. Up until today these interesting molecules could only be produced using an engineered Starmerella bombicola strain (∆ugtB1::URA3 G9) producing GLs instead of sophorolipids, albeit with a very low average productivity (0.01 g/L/h). In this study, we investigated the reason(s) for this via reverse-transcription quantitative polymerase chain reaction and Liquid chromatography-multireaction monitoring-mass spectrometry. We found that all glycolipid biosynthetic genes and enzymes were downregulated in the ∆ugtB1 G9 strain in comparison to the wild type. The underlying reason for this downregulation was further investigated by performing quantitative metabolome comparison of the ∆ugtB1 G9 strain with the wild type and two other engineered strains also tinkered in their glycolipid biosynthetic gene cluster. This analysis revealed a clear distortion of the entire metabolism of the ∆ugtB1 G9 strain compared to all the other strains. Because the parental strain of the former was a spontaneous ∆ura3 mutant potentially containing other "hidden" mutations, a new GL production strain was generated based on a rationally engineered ∆ura3 mutant (PT36). Indeed, a 50-fold GL productivity increase (0.51 g/L/h) was obtained with the new ∆ugtB1::URA3 PT36 strain compared with the G9-based strain (0.01 g/L/h) in a 10 L bioreactor experiment, yielding 118 g/L GLs instead of 8.39 g/L. Purification was investigated and basic properties of the purified GLs were determined. This study forms the base for further development and optimization of S. bombicola as a production platform strain for (new) biochemicals.
fermentation, purification, glucolipid, Starmerella bombicola, strain engineering, (bio)surfactants
Structure type: monomer
Location inside paper: Fig. 1, non- and mono acetylated glycolipid
Compound class: glycolipid
Contained glycoepitopes: IEDB_142488,IEDB_146664,IEDB_983931,SB_192
Methods: ESI-MS, enzymatic digestion, extraction, LC-MS, cell growth, LC-MS/MS, precipitation, sonication, antimicrobial assay, centrifugation, qRT-PCR, optical density measurement, UPLC-ELSD, RPLC
Enzymes that release or process the structure: cyp52m1 (monooxygenase), ugta1 (glucosyltransferase), ugtb1 (glucosyltransferase), At (acetyltransferase), sble
Biosynthesis and genetic data: biochemical data, genetic data
Related record ID(s): 50966
NCBI Taxonomy refs (TaxIDs): 75736
Show glycosyltransferases
There is only one chemically distinct structure:
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