Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese Making
In traditional cheese making, pregastric lipolytic enzymes of animal origin are used for the acceleration of ripening and the formation of spicy flavor compounds. Especially for cheese specialities, such as Pecorino, Provolone, or Feta, pregastric esterases (PGE) play an important role. A lipase fro...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-08-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/11/17/2608 |
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| author | Niklas Broel Miriam A. Sowa Julia Manhard Alexander Siegl Edgar Weichhard Holger Zorn Binglin Li Martin Gand |
| author_facet | Niklas Broel Miriam A. Sowa Julia Manhard Alexander Siegl Edgar Weichhard Holger Zorn Binglin Li Martin Gand |
| author_sort | Niklas Broel |
| collection | DOAJ |
| description | In traditional cheese making, pregastric lipolytic enzymes of animal origin are used for the acceleration of ripening and the formation of spicy flavor compounds. Especially for cheese specialities, such as Pecorino, Provolone, or Feta, pregastric esterases (PGE) play an important role. A lipase from <i>Pleurotus citrinopileatus</i> could serve as a substitute for these animal-derived enzymes, thus offering vegetarian, kosher, and halal alternatives. However, the hydrolytic activity of this enzyme towards long-chain fatty acids is slightly too high, which may lead to off-flavors during long-term ripening. Therefore, an optimization via protein engineering (PE) was performed by changing the specificity towards medium-chain fatty acids. With a semi-rational design, possible mutants at eight different positions were created and analyzed in silico. Heterologous expression was performed for 24 predicted mutants, of which 18 caused a change in the hydrolysis profile. Three mutants (F91L, L302G, and L305A) were used in application tests to produce Feta-type brine cheese. The sensory analyses showed promising results for cheeses prepared with the L305A mutant, and SPME-GC-MS analysis of volatile free fatty acids supported these findings. Therefore, altering the chain length specificity via PE becomes a powerful tool for the replacement of PGEs in cheese making. |
| first_indexed | 2024-03-10T01:48:47Z |
| format | Article |
| id | doaj.art-2133d4e25ba046479e5c8a8b19aeda65 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-10T01:48:47Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-2133d4e25ba046479e5c8a8b19aeda652023-11-23T13:08:12ZengMDPI AGFoods2304-81582022-08-011117260810.3390/foods11172608Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese MakingNiklas Broel0Miriam A. Sowa1Julia Manhard2Alexander Siegl3Edgar Weichhard4Holger Zorn5Binglin Li6Martin Gand7Institute of Food Chemistry and Food Biotechnology, Justus Liebig University Giessen, 35392 Giessen, GermanyInstitute of Food Chemistry and Food Biotechnology, Justus Liebig University Giessen, 35392 Giessen, GermanyOptiferm GmbH, 87466 Oy-Mittelberg, GermanyOptiferm GmbH, 87466 Oy-Mittelberg, GermanyOptiferm GmbH, 87466 Oy-Mittelberg, GermanyInstitute of Food Chemistry and Food Biotechnology, Justus Liebig University Giessen, 35392 Giessen, GermanyCollege of Food Science and Engineering, Northwest University, Xi’an 710069, ChinaInstitute of Food Chemistry and Food Biotechnology, Justus Liebig University Giessen, 35392 Giessen, GermanyIn traditional cheese making, pregastric lipolytic enzymes of animal origin are used for the acceleration of ripening and the formation of spicy flavor compounds. Especially for cheese specialities, such as Pecorino, Provolone, or Feta, pregastric esterases (PGE) play an important role. A lipase from <i>Pleurotus citrinopileatus</i> could serve as a substitute for these animal-derived enzymes, thus offering vegetarian, kosher, and halal alternatives. However, the hydrolytic activity of this enzyme towards long-chain fatty acids is slightly too high, which may lead to off-flavors during long-term ripening. Therefore, an optimization via protein engineering (PE) was performed by changing the specificity towards medium-chain fatty acids. With a semi-rational design, possible mutants at eight different positions were created and analyzed in silico. Heterologous expression was performed for 24 predicted mutants, of which 18 caused a change in the hydrolysis profile. Three mutants (F91L, L302G, and L305A) were used in application tests to produce Feta-type brine cheese. The sensory analyses showed promising results for cheeses prepared with the L305A mutant, and SPME-GC-MS analysis of volatile free fatty acids supported these findings. Therefore, altering the chain length specificity via PE becomes a powerful tool for the replacement of PGEs in cheese making.https://www.mdpi.com/2304-8158/11/17/2608<i>Pleurotus citrinopileatus</i>lipasesemi-rational designcheese makingchain length specificity |
| spellingShingle | Niklas Broel Miriam A. Sowa Julia Manhard Alexander Siegl Edgar Weichhard Holger Zorn Binglin Li Martin Gand Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese Making Foods <i>Pleurotus citrinopileatus</i> lipase semi-rational design cheese making chain length specificity |
| title | Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese Making |
| title_full | Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese Making |
| title_fullStr | Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese Making |
| title_full_unstemmed | Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese Making |
| title_short | Altering the Chain Length Specificity of a Lipase from <i>Pleurotus citrinopileatus</i> for the Application in Cheese Making |
| title_sort | altering the chain length specificity of a lipase from i pleurotus citrinopileatus i for the application in cheese making |
| topic | <i>Pleurotus citrinopileatus</i> lipase semi-rational design cheese making chain length specificity |
| url | https://www.mdpi.com/2304-8158/11/17/2608 |
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