The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LC
Freeze drying is a commonly used method for preserving probiotic bacteria and live biotherapeutic products. Before drying, the bacterial cells are formulated with a lyoprotectant, and the design of these two process steps are crucial to achieve a high-quality product. There are several factors that...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-12-01
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| Series: | Applied Microbiology |
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| Online Access: | https://www.mdpi.com/2673-8007/3/4/92 |
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| author | Nisha Tyagi Zandra Gidlöf Daniel Tristan Osanlóo Elizabeth S. Collier Sandeep Kadekar Lovisa Ringstad Anna Millqvist Fureby Stefan Roos |
| author_facet | Nisha Tyagi Zandra Gidlöf Daniel Tristan Osanlóo Elizabeth S. Collier Sandeep Kadekar Lovisa Ringstad Anna Millqvist Fureby Stefan Roos |
| author_sort | Nisha Tyagi |
| collection | DOAJ |
| description | Freeze drying is a commonly used method for preserving probiotic bacteria and live biotherapeutic products. Before drying, the bacterial cells are formulated with a lyoprotectant, and the design of these two process steps are crucial to achieve a high-quality product. There are several factors that may affect the biological and physicochemical properties of the freeze-dried cells and we have used a Design of Experiment approach to investigate the effects of formulation and freeze-drying parameters on properties and performance of <i>Limosilactobacillus reuteri</i> R2LC. The biological characteristics of the dried bacteria were evaluated by measuring cell survival, metabolic activity and stability, and physicochemical characteristics were studied using visual inspection, differential scanning calorimetry (DSC), scanning electron microscopy (SEM), and analysis of residual moisture content and bacterial aggregation. A comparison between the lyoprotectants trehalose and sucrose showed that the latter gave better freeze-drying survival, metabolic activity, and storage stability. We also want to highlight that there was a correlation between bacterial concentration, metabolic activity, and aggregation of bacteria, where a higher concentration (10<sup>10</sup> CFU/mL) resulted in both higher metabolic activity and aggregation. Several other process and formulation factors affected both the biological and physicochemical properties of freeze-dried <i>L. reuteri</i> R2LC and it could be concluded that care must be taken to develop a production method that generates a product with high and consistent quality. These results may, or may not, be strain specific. |
| first_indexed | 2024-03-08T18:35:10Z |
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| id | doaj.art-c6e5ff2042554e4b824e1b041b6c8249 |
| institution | Directory Open Access Journal |
| issn | 2673-8007 |
| language | English |
| last_indexed | 2024-03-08T18:35:10Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Applied Microbiology |
| spelling | doaj.art-c6e5ff2042554e4b824e1b041b6c82492023-12-29T15:46:07ZengMDPI AGApplied Microbiology2673-80072023-12-01341370138710.3390/applmicrobiol3040092The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LCNisha Tyagi0Zandra Gidlöf1Daniel Tristan Osanlóo2Elizabeth S. Collier3Sandeep Kadekar4Lovisa Ringstad5Anna Millqvist Fureby6Stefan Roos7Department of Molecular Sciences, Uppsala BioCenter, Swedish University of Agricultural Sciences, 750 07 Uppsala, SwedenRISE Research Institutes of Sweden, 114 28 Stockholm, SwedenRISE Research Institutes of Sweden, 114 28 Stockholm, SwedenRISE Research Institutes of Sweden, 114 28 Stockholm, SwedenIlya Pharma AB, Dag Hammarskjölds väg 30, 752 37 Uppsala, SwedenIlya Pharma AB, Dag Hammarskjölds väg 30, 752 37 Uppsala, SwedenRISE Research Institutes of Sweden, 114 28 Stockholm, SwedenDepartment of Molecular Sciences, Uppsala BioCenter, Swedish University of Agricultural Sciences, 750 07 Uppsala, SwedenFreeze drying is a commonly used method for preserving probiotic bacteria and live biotherapeutic products. Before drying, the bacterial cells are formulated with a lyoprotectant, and the design of these two process steps are crucial to achieve a high-quality product. There are several factors that may affect the biological and physicochemical properties of the freeze-dried cells and we have used a Design of Experiment approach to investigate the effects of formulation and freeze-drying parameters on properties and performance of <i>Limosilactobacillus reuteri</i> R2LC. The biological characteristics of the dried bacteria were evaluated by measuring cell survival, metabolic activity and stability, and physicochemical characteristics were studied using visual inspection, differential scanning calorimetry (DSC), scanning electron microscopy (SEM), and analysis of residual moisture content and bacterial aggregation. A comparison between the lyoprotectants trehalose and sucrose showed that the latter gave better freeze-drying survival, metabolic activity, and storage stability. We also want to highlight that there was a correlation between bacterial concentration, metabolic activity, and aggregation of bacteria, where a higher concentration (10<sup>10</sup> CFU/mL) resulted in both higher metabolic activity and aggregation. Several other process and formulation factors affected both the biological and physicochemical properties of freeze-dried <i>L. reuteri</i> R2LC and it could be concluded that care must be taken to develop a production method that generates a product with high and consistent quality. These results may, or may not, be strain specific.https://www.mdpi.com/2673-8007/3/4/92probioticslive biotherapeutics<i>Limosilactobacillus reuteri</i> R2LCformulationfreeze-dryingmetabolic activity |
| spellingShingle | Nisha Tyagi Zandra Gidlöf Daniel Tristan Osanlóo Elizabeth S. Collier Sandeep Kadekar Lovisa Ringstad Anna Millqvist Fureby Stefan Roos The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LC Applied Microbiology probiotics live biotherapeutics <i>Limosilactobacillus reuteri</i> R2LC formulation freeze-drying metabolic activity |
| title | The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LC |
| title_full | The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LC |
| title_fullStr | The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LC |
| title_full_unstemmed | The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LC |
| title_short | The Impact of Formulation and Freeze Drying on the Properties and Performance of Freeze-Dried <i>Limosilactobacillus reuteri</i> R2LC |
| title_sort | impact of formulation and freeze drying on the properties and performance of freeze dried i limosilactobacillus reuteri i r2lc |
| topic | probiotics live biotherapeutics <i>Limosilactobacillus reuteri</i> R2LC formulation freeze-drying metabolic activity |
| url | https://www.mdpi.com/2673-8007/3/4/92 |
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