Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates
Abstract Background One critical parameter in microbial cultivations is the composition of the cultivation medium. Nowadays, the application of chemically defined media increases, due to a more defined and reproducible fermentation performance than in complex media. In order, to improve cost-effecti...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-07-01
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| Series: | BMC Biotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12896-023-00793-7 |
| _version_ | 1797769424430694400 |
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| author | Jennifer Goldmanns Georg Andreas Röhling Marie Kristine Lipa Theresa Scholand Alexander Deitert Tobias May Evangeline Priya Haas Matthias Boy Andrea Herold Jochen Büchs |
| author_facet | Jennifer Goldmanns Georg Andreas Röhling Marie Kristine Lipa Theresa Scholand Alexander Deitert Tobias May Evangeline Priya Haas Matthias Boy Andrea Herold Jochen Büchs |
| author_sort | Jennifer Goldmanns |
| collection | DOAJ |
| description | Abstract Background One critical parameter in microbial cultivations is the composition of the cultivation medium. Nowadays, the application of chemically defined media increases, due to a more defined and reproducible fermentation performance than in complex media. In order, to improve cost-effectiveness of fermentation processes using chemically defined media, the media should not contain nutrients in large excess. Additionally, to obtain high product yields, the nutrient concentrations should not be limiting. Therefore, efficient medium optimization techniques are required which adapt medium compositions to the specific nutrient requirements of microorganisms. Results Since most Paenibacillus cultivation protocols so far described in literature are based on complex ingredients, in this study, a chemically defined medium for an industrially relevant Paenibacillus polymyxa strain was developed. A recently reported method, which combines a systematic experimental procedure in combination with online monitoring of the respiration activity, was applied and extended to identify growth limitations for Paenibacillus polymyxa. All cultivations were performed in microtiter plates. By systematically increasing the concentrations of different nutrient groups, nicotinic acid was identified as a growth-limiting component. Additionally, an insufficient buffer capacity was observed. After optimizing the growth in the chemically defined medium, the medium components were systematically reduced to contain only nutrients relevant for growth. Vitamins were reduced to nicotinic acid and biotin, and amino acids to methionine, histidine, proline, arginine, and glutamate. Nucleobases/-sides could be completely left out of the medium. Finally, the cultivation in the reduced medium was reproduced in a laboratory fermenter. Conclusion In this study, a reliable and time-efficient high-throughput methodology was extended to investigate limitations in chemically defined media. The interpretation of online measured respiration activities agreed well with the growth performance of samples measured in parallel via offline analyses. Furthermore, the cultivation in microtiter plates was validated in a laboratory fermenter. The results underline the benefits of online monitoring of the respiration activity already in the early stages of process development, to avoid limitations of medium components, oxygen limitation and pH inhibition during the scale-up. |
| first_indexed | 2024-03-12T21:08:49Z |
| format | Article |
| id | doaj.art-3b613fbdea584f7e826514e691495b96 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-03-12T21:08:49Z |
| publishDate | 2023-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-3b613fbdea584f7e826514e691495b962023-07-30T11:18:44ZengBMCBMC Biotechnology1472-67502023-07-0123112310.1186/s12896-023-00793-7Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter platesJennifer Goldmanns0Georg Andreas Röhling1Marie Kristine Lipa2Theresa Scholand3Alexander Deitert4Tobias May5Evangeline Priya Haas6Matthias Boy7Andrea Herold8Jochen Büchs9RWTH Aachen University, AVT – Biochemical EngineeringRWTH Aachen University, AVT – Biochemical EngineeringRWTH Aachen University, AVT – Biochemical EngineeringRWTH Aachen University, AVT – Biochemical EngineeringRWTH Aachen University, AVT – Biochemical EngineeringBASF SEBASF SEBASF SEBASF SERWTH Aachen University, AVT – Biochemical EngineeringAbstract Background One critical parameter in microbial cultivations is the composition of the cultivation medium. Nowadays, the application of chemically defined media increases, due to a more defined and reproducible fermentation performance than in complex media. In order, to improve cost-effectiveness of fermentation processes using chemically defined media, the media should not contain nutrients in large excess. Additionally, to obtain high product yields, the nutrient concentrations should not be limiting. Therefore, efficient medium optimization techniques are required which adapt medium compositions to the specific nutrient requirements of microorganisms. Results Since most Paenibacillus cultivation protocols so far described in literature are based on complex ingredients, in this study, a chemically defined medium for an industrially relevant Paenibacillus polymyxa strain was developed. A recently reported method, which combines a systematic experimental procedure in combination with online monitoring of the respiration activity, was applied and extended to identify growth limitations for Paenibacillus polymyxa. All cultivations were performed in microtiter plates. By systematically increasing the concentrations of different nutrient groups, nicotinic acid was identified as a growth-limiting component. Additionally, an insufficient buffer capacity was observed. After optimizing the growth in the chemically defined medium, the medium components were systematically reduced to contain only nutrients relevant for growth. Vitamins were reduced to nicotinic acid and biotin, and amino acids to methionine, histidine, proline, arginine, and glutamate. Nucleobases/-sides could be completely left out of the medium. Finally, the cultivation in the reduced medium was reproduced in a laboratory fermenter. Conclusion In this study, a reliable and time-efficient high-throughput methodology was extended to investigate limitations in chemically defined media. The interpretation of online measured respiration activities agreed well with the growth performance of samples measured in parallel via offline analyses. Furthermore, the cultivation in microtiter plates was validated in a laboratory fermenter. The results underline the benefits of online monitoring of the respiration activity already in the early stages of process development, to avoid limitations of medium components, oxygen limitation and pH inhibition during the scale-up.https://doi.org/10.1186/s12896-023-00793-7Paenibacillus polymyxaChemically defined mediaOnline monitoringMedium optimizationGrowth limitations |
| spellingShingle | Jennifer Goldmanns Georg Andreas Röhling Marie Kristine Lipa Theresa Scholand Alexander Deitert Tobias May Evangeline Priya Haas Matthias Boy Andrea Herold Jochen Büchs Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates BMC Biotechnology Paenibacillus polymyxa Chemically defined media Online monitoring Medium optimization Growth limitations |
| title | Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates |
| title_full | Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates |
| title_fullStr | Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates |
| title_full_unstemmed | Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates |
| title_short | Development of a chemically defined medium for Paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates |
| title_sort | development of a chemically defined medium for paenibacillus polymyxa by parallel online monitoring of the respiration activity in microtiter plates |
| topic | Paenibacillus polymyxa Chemically defined media Online monitoring Medium optimization Growth limitations |
| url | https://doi.org/10.1186/s12896-023-00793-7 |
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