State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i>
Oxidoreductase (OXR) enzymes are in high demand for biocatalytic applications in the food industry and cosmetics (glucose oxidase (GOx) and cellobiose dehydrogenase (CDH)), bioremediations (horseradish peroxidase (HRP) and laccase (LAC)), and medicine for biosensors and miniature biofuel cells (GOx,...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/10/2/93 |
| _version_ | 1797298260854964224 |
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| author | Milica Crnoglavac Popović Marija Stanišić Radivoje Prodanović |
| author_facet | Milica Crnoglavac Popović Marija Stanišić Radivoje Prodanović |
| author_sort | Milica Crnoglavac Popović |
| collection | DOAJ |
| description | Oxidoreductase (OXR) enzymes are in high demand for biocatalytic applications in the food industry and cosmetics (glucose oxidase (GOx) and cellobiose dehydrogenase (CDH)), bioremediations (horseradish peroxidase (HRP) and laccase (LAC)), and medicine for biosensors and miniature biofuel cells (GOx, CDH, LAC, and HRP). They can be used in a soluble form and/or within the yeast cell walls expressed as chimeras on the surface of yeast cells (YSD), such as <i>P. pastoris</i> and <i>S. cerevisiae</i>. However, most of the current studies suffer from either low yield for soluble enzyme expression or low enzyme activity when expressed as chimeric proteins using YSD. This is always the case in studies dealing with the heterologous expression of oxidoreductase enzymes, since there is a requirement not only for multiple OXR gene integrations into the yeast genome (super transformations), and codon optimization, but also very careful design of fermentation media composition and fermentation conditions during expression due to the need for transition metals (copper and iron) and metabolic precursors of FAD and heme. Therefore, scientists are still trying to find the optimal formula using the above-mentioned approaches; most recently, researcher started using protein engineering and directed evolution to increase in the yield of recombinant enzyme production. In this review article, we will cover all the current state-of-the-art technologies and most recent advances in the field that yielded a high expression level for some of these enzymes in specially designed expression/fermentation systems. We will also tackle and discuss new possibilities for further increases in fermentation yield using cutting-edge technologies such as directed evolution, protein and strain engineering, high-throughput screening methods based on in vitro compartmentalization, flow cytometry, and microfluidics. |
| first_indexed | 2024-03-07T22:33:22Z |
| format | Article |
| id | doaj.art-29c4ff33535645ce9bc0be5fe7a096e5 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-07T22:33:22Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-29c4ff33535645ce9bc0be5fe7a096e52024-02-23T15:15:55ZengMDPI AGFermentation2311-56372024-02-011029310.3390/fermentation10020093State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i>Milica Crnoglavac Popović0Marija Stanišić1Radivoje Prodanović2Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, SerbiaFaculty of Chemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, SerbiaFaculty of Chemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, SerbiaOxidoreductase (OXR) enzymes are in high demand for biocatalytic applications in the food industry and cosmetics (glucose oxidase (GOx) and cellobiose dehydrogenase (CDH)), bioremediations (horseradish peroxidase (HRP) and laccase (LAC)), and medicine for biosensors and miniature biofuel cells (GOx, CDH, LAC, and HRP). They can be used in a soluble form and/or within the yeast cell walls expressed as chimeras on the surface of yeast cells (YSD), such as <i>P. pastoris</i> and <i>S. cerevisiae</i>. However, most of the current studies suffer from either low yield for soluble enzyme expression or low enzyme activity when expressed as chimeric proteins using YSD. This is always the case in studies dealing with the heterologous expression of oxidoreductase enzymes, since there is a requirement not only for multiple OXR gene integrations into the yeast genome (super transformations), and codon optimization, but also very careful design of fermentation media composition and fermentation conditions during expression due to the need for transition metals (copper and iron) and metabolic precursors of FAD and heme. Therefore, scientists are still trying to find the optimal formula using the above-mentioned approaches; most recently, researcher started using protein engineering and directed evolution to increase in the yield of recombinant enzyme production. In this review article, we will cover all the current state-of-the-art technologies and most recent advances in the field that yielded a high expression level for some of these enzymes in specially designed expression/fermentation systems. We will also tackle and discuss new possibilities for further increases in fermentation yield using cutting-edge technologies such as directed evolution, protein and strain engineering, high-throughput screening methods based on in vitro compartmentalization, flow cytometry, and microfluidics.https://www.mdpi.com/2311-5637/10/2/93recombinantoxidoreductaseexpressionyeastsdirected evolutionhigh-throughput screening |
| spellingShingle | Milica Crnoglavac Popović Marija Stanišić Radivoje Prodanović State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i> Fermentation recombinant oxidoreductase expression yeasts directed evolution high-throughput screening |
| title | State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i> |
| title_full | State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i> |
| title_fullStr | State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i> |
| title_full_unstemmed | State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i> |
| title_short | State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts <i>P. pastoris</i> and <i>S. cerevisiae</i> |
| title_sort | state of the art technologies for high yield heterologous expression and production of oxidoreductase enzymes glucose oxidase cellobiose dehydrogenase horseradish peroxidase and laccases in yeasts i p pastoris i and i s cerevisiae i |
| topic | recombinant oxidoreductase expression yeasts directed evolution high-throughput screening |
| url | https://www.mdpi.com/2311-5637/10/2/93 |
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