Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic Effect
In this work, the trisaccharide isomelezitose was overproduced from sucrose using a biocatalyst based on immobilized <i>Escherichia coli</i> cells harbouring the α-glucosidase from the yeast <i>Metschnikowia reukaufii</i>, the best native producer of this sugar described to d...
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| Format: | Article |
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MDPI AG
2022-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/20/12682 |
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| author | Martin Garcia-Gonzalez Fadia V. Cervantes Ricardo P. Ipiales Angeles de la Rubia Francisco J. Plou María Fernández-Lobato |
| author_facet | Martin Garcia-Gonzalez Fadia V. Cervantes Ricardo P. Ipiales Angeles de la Rubia Francisco J. Plou María Fernández-Lobato |
| author_sort | Martin Garcia-Gonzalez |
| collection | DOAJ |
| description | In this work, the trisaccharide isomelezitose was overproduced from sucrose using a biocatalyst based on immobilized <i>Escherichia coli</i> cells harbouring the α-glucosidase from the yeast <i>Metschnikowia reukaufii</i>, the best native producer of this sugar described to date. The overall process for isomelezitose production and purification was performed in three simple steps: (i) oligosaccharides synthesis by alginate-entrapped <i>E. coli</i>; (ii) elimination of monosaccharides (glucose and fructose) using alginate-entrapped <i>Komagataella phaffii</i> cells; and (iii) semi-preparative high performance liquid chromatography under isocratic conditions. As result, approximately 2.15 g of isomelezitose (purity exceeding 95%) was obtained from 15 g of sucrose. The potential prebiotic effect of this sugar on probiotic bacteria (<i>Lactobacillus casei</i>, <i>Lactobacillus rhamnosus</i> and <i>Enterococcus faecium</i>) was analysed using in vitro assays for the first time. The growth of all probiotic bacteria cultures supplemented with isomelezitose was significantly improved and was similar to that of cultures supplemented with a commercial mixture of fructo-oligosaccharides. In addition, when isomelezitose was added to the bacteria cultures, the production of organic acids (mainly butyrate) was significantly promoted. Therefore, these results confirm that isomelezitose is a potential novel prebiotic that could be included in healthier foodstuffs designed for human gastrointestinal balance maintenance. |
| first_indexed | 2024-03-09T20:04:56Z |
| format | Article |
| id | doaj.art-88fad6cdc92145d8a04c4efe10eeaef0 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T20:04:56Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-88fad6cdc92145d8a04c4efe10eeaef02023-11-24T00:35:18ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-10-0123201268210.3390/ijms232012682Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic EffectMartin Garcia-Gonzalez0Fadia V. Cervantes1Ricardo P. Ipiales2Angeles de la Rubia3Francisco J. Plou4María Fernández-Lobato5Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma de Madrid, C/Nicolás Cabrera, 1, 28049 Madrid, SpainInstituto de Catálisis y Petroleoquímica, CSIC, 28049 Madrid, SpainDepartamento de Ingeniería Química, Universidad Autónoma de Madrid (UAM), Campus de Cantoblanco, 28049 Madrid, SpainDepartamento de Ingeniería Química, Universidad Autónoma de Madrid (UAM), Campus de Cantoblanco, 28049 Madrid, SpainInstituto de Catálisis y Petroleoquímica, CSIC, 28049 Madrid, SpainDepartamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma de Madrid, C/Nicolás Cabrera, 1, 28049 Madrid, SpainIn this work, the trisaccharide isomelezitose was overproduced from sucrose using a biocatalyst based on immobilized <i>Escherichia coli</i> cells harbouring the α-glucosidase from the yeast <i>Metschnikowia reukaufii</i>, the best native producer of this sugar described to date. The overall process for isomelezitose production and purification was performed in three simple steps: (i) oligosaccharides synthesis by alginate-entrapped <i>E. coli</i>; (ii) elimination of monosaccharides (glucose and fructose) using alginate-entrapped <i>Komagataella phaffii</i> cells; and (iii) semi-preparative high performance liquid chromatography under isocratic conditions. As result, approximately 2.15 g of isomelezitose (purity exceeding 95%) was obtained from 15 g of sucrose. The potential prebiotic effect of this sugar on probiotic bacteria (<i>Lactobacillus casei</i>, <i>Lactobacillus rhamnosus</i> and <i>Enterococcus faecium</i>) was analysed using in vitro assays for the first time. The growth of all probiotic bacteria cultures supplemented with isomelezitose was significantly improved and was similar to that of cultures supplemented with a commercial mixture of fructo-oligosaccharides. In addition, when isomelezitose was added to the bacteria cultures, the production of organic acids (mainly butyrate) was significantly promoted. Therefore, these results confirm that isomelezitose is a potential novel prebiotic that could be included in healthier foodstuffs designed for human gastrointestinal balance maintenance.https://www.mdpi.com/1422-0067/23/20/12682isomelezitoseimmobilized cellsprebiotic agent<i>Lactobacillus casei</i><i>Lactobacillus rhamnosus</i><i>Enterococcus faecium</i> |
| spellingShingle | Martin Garcia-Gonzalez Fadia V. Cervantes Ricardo P. Ipiales Angeles de la Rubia Francisco J. Plou María Fernández-Lobato Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic Effect International Journal of Molecular Sciences isomelezitose immobilized cells prebiotic agent <i>Lactobacillus casei</i> <i>Lactobacillus rhamnosus</i> <i>Enterococcus faecium</i> |
| title | Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic Effect |
| title_full | Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic Effect |
| title_fullStr | Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic Effect |
| title_full_unstemmed | Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic Effect |
| title_short | Isomelezitose Overproduction by Alginate-Entrapped Recombinant <i>E. coli</i> Cells and In Vitro Evaluation of Its Potential Prebiotic Effect |
| title_sort | isomelezitose overproduction by alginate entrapped recombinant i e coli i cells and in vitro evaluation of its potential prebiotic effect |
| topic | isomelezitose immobilized cells prebiotic agent <i>Lactobacillus casei</i> <i>Lactobacillus rhamnosus</i> <i>Enterococcus faecium</i> |
| url | https://www.mdpi.com/1422-0067/23/20/12682 |
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