Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants
The aim of the present work was to obtain microbial lipids (single-cell oils and SCOs) from oleaginous yeast cultivated on biodiesel-derived glycerol and subsequently proceed to the enzymatic synthesis of high-value biosurfactant-type molecules in an aqueous medium, with SCOs implicated as acyl dono...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-12-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/1/714 |
| _version_ | 1797625584290889728 |
|---|---|
| author | Dimitris Karayannis Seraphim Papanikolaou Christos Vatistas Cédric Paris Isabelle Chevalot |
| author_facet | Dimitris Karayannis Seraphim Papanikolaou Christos Vatistas Cédric Paris Isabelle Chevalot |
| author_sort | Dimitris Karayannis |
| collection | DOAJ |
| description | The aim of the present work was to obtain microbial lipids (single-cell oils and SCOs) from oleaginous yeast cultivated on biodiesel-derived glycerol and subsequently proceed to the enzymatic synthesis of high-value biosurfactant-type molecules in an aqueous medium, with SCOs implicated as acyl donors (ADs). Indeed, the initial screening of five non-conventional oleaginous yeasts revealed that the most important lipid producer was the microorganism <i>Cryptococcus curvatus</i> ATCC 20509. SCO production was optimised according to the nature of the nitrogen source and the initial concentration of glycerol (Glyc0) employed in the medium. Lipids up to 50% <i>w</i>/<i>w</i> in dry cell weight (DCW) (SCO<sub>max</sub> = 6.1 g/L) occurred at Glyc0 ≈ 70 g/L (C/N ≈ 80 moles/moles). Thereafter, lipids were recovered and were subsequently used as ADs in the N-acylation reaction catalysed by aminoacylases produced from <i>Streptomyces ambofaciens</i> ATCC 23877 under aqueous conditions, while <i>Candida antarctica</i> lipase B (CALB) was used as a reference enzyme. Aminoacylases revealed excellent activity towards the synthesis of acyl-lysine only when free fatty acids (FAs) were used as the AD, and the rare regioselectivity in the α-amino group, which has a great impact on the preservation of the functional side chains of any amino acids or peptides. Aminoacylases presented higher α-oleoyl-lysine productivity and final titer (8.3 g/L) with hydrolysed SCO than with hydrolysed vegetable oil. The substrate specificity of both enzymes towards the three main FAs found in SCO was studied, and a new parameter was defined, viz., Specificity factor (Sf), which expresses the relative substrate specificity of an enzyme towards a FA present in a FA mixture. The Sf value of aminoacylases was the highest with palmitic acid in all cases tested, ranging from 2.0 to 3.0, while that of CALB was with linoleic acid (0.9–1.5). To the best of our knowledge, this is the first time that a microbial oil has been successfully used as AD for biosurfactant synthesis. This bio-refinery approach illustrates the concept of a state-of-the-art combination of enzyme and microbial technology to produce high-value biosurfactants through environmentally friendly and economically sound processes. |
| first_indexed | 2024-03-11T09:58:26Z |
| format | Article |
| id | doaj.art-f88a2cf357884c648071414d7ea864cf |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T09:58:26Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f88a2cf357884c648071414d7ea864cf2023-11-16T15:37:21ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0124171410.3390/ijms24010714Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based BiosurfactantsDimitris Karayannis0Seraphim Papanikolaou1Christos Vatistas2Cédric Paris3Isabelle Chevalot4Reaction and Chemical Engineering Laboratory, University of Lorraine, F-54000 Nancy, FranceReaction and Chemical Engineering Laboratory, University of Lorraine, F-54000 Nancy, FranceDepartment of Food Science & Human Nutrition, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, GreeceReaction and Chemical Engineering Laboratory, University of Lorraine, F-54000 Nancy, FranceReaction and Chemical Engineering Laboratory, University of Lorraine, F-54000 Nancy, FranceThe aim of the present work was to obtain microbial lipids (single-cell oils and SCOs) from oleaginous yeast cultivated on biodiesel-derived glycerol and subsequently proceed to the enzymatic synthesis of high-value biosurfactant-type molecules in an aqueous medium, with SCOs implicated as acyl donors (ADs). Indeed, the initial screening of five non-conventional oleaginous yeasts revealed that the most important lipid producer was the microorganism <i>Cryptococcus curvatus</i> ATCC 20509. SCO production was optimised according to the nature of the nitrogen source and the initial concentration of glycerol (Glyc0) employed in the medium. Lipids up to 50% <i>w</i>/<i>w</i> in dry cell weight (DCW) (SCO<sub>max</sub> = 6.1 g/L) occurred at Glyc0 ≈ 70 g/L (C/N ≈ 80 moles/moles). Thereafter, lipids were recovered and were subsequently used as ADs in the N-acylation reaction catalysed by aminoacylases produced from <i>Streptomyces ambofaciens</i> ATCC 23877 under aqueous conditions, while <i>Candida antarctica</i> lipase B (CALB) was used as a reference enzyme. Aminoacylases revealed excellent activity towards the synthesis of acyl-lysine only when free fatty acids (FAs) were used as the AD, and the rare regioselectivity in the α-amino group, which has a great impact on the preservation of the functional side chains of any amino acids or peptides. Aminoacylases presented higher α-oleoyl-lysine productivity and final titer (8.3 g/L) with hydrolysed SCO than with hydrolysed vegetable oil. The substrate specificity of both enzymes towards the three main FAs found in SCO was studied, and a new parameter was defined, viz., Specificity factor (Sf), which expresses the relative substrate specificity of an enzyme towards a FA present in a FA mixture. The Sf value of aminoacylases was the highest with palmitic acid in all cases tested, ranging from 2.0 to 3.0, while that of CALB was with linoleic acid (0.9–1.5). To the best of our knowledge, this is the first time that a microbial oil has been successfully used as AD for biosurfactant synthesis. This bio-refinery approach illustrates the concept of a state-of-the-art combination of enzyme and microbial technology to produce high-value biosurfactants through environmentally friendly and economically sound processes.https://www.mdpi.com/1422-0067/24/1/714biosurfactantmicrobial lipidaminoacylasesN-acylationspecificity factor |
| spellingShingle | Dimitris Karayannis Seraphim Papanikolaou Christos Vatistas Cédric Paris Isabelle Chevalot Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants International Journal of Molecular Sciences biosurfactant microbial lipid aminoacylases N-acylation specificity factor |
| title | Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants |
| title_full | Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants |
| title_fullStr | Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants |
| title_full_unstemmed | Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants |
| title_short | Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants |
| title_sort | yeast lipid produced through glycerol conversions and its use for enzymatic synthesis of amino acid based biosurfactants |
| topic | biosurfactant microbial lipid aminoacylases N-acylation specificity factor |
| url | https://www.mdpi.com/1422-0067/24/1/714 |
| work_keys_str_mv | AT dimitriskarayannis yeastlipidproducedthroughglycerolconversionsanditsuseforenzymaticsynthesisofaminoacidbasedbiosurfactants AT seraphimpapanikolaou yeastlipidproducedthroughglycerolconversionsanditsuseforenzymaticsynthesisofaminoacidbasedbiosurfactants AT christosvatistas yeastlipidproducedthroughglycerolconversionsanditsuseforenzymaticsynthesisofaminoacidbasedbiosurfactants AT cedricparis yeastlipidproducedthroughglycerolconversionsanditsuseforenzymaticsynthesisofaminoacidbasedbiosurfactants AT isabellechevalot yeastlipidproducedthroughglycerolconversionsanditsuseforenzymaticsynthesisofaminoacidbasedbiosurfactants |