Cargando…
Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors
BACKGROUND: Conventional experiments in small scale are often performed in a ‘Black Box’ fashion, analyzing only the product concentration in the final sample. Online monitoring of relevant process characteristics and parameters such as substrate limitation, product inhibition and oxygen supply is l...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4531433/ https://www.ncbi.nlm.nih.gov/pubmed/26265936 http://dx.doi.org/10.1186/s13036-015-0005-0 |
_version_ | 1782385034733289472 |
---|---|
author | Wewetzer, S. J. Kunze, M. Ladner, T. Luchterhand, B. Roth, S. Rahmen, N. Kloß, R. Costa e Silva, A. Regestein, L. Büchs, J. |
author_facet | Wewetzer, S. J. Kunze, M. Ladner, T. Luchterhand, B. Roth, S. Rahmen, N. Kloß, R. Costa e Silva, A. Regestein, L. Büchs, J. |
author_sort | Wewetzer, S. J. |
collection | PubMed |
description | BACKGROUND: Conventional experiments in small scale are often performed in a ‘Black Box’ fashion, analyzing only the product concentration in the final sample. Online monitoring of relevant process characteristics and parameters such as substrate limitation, product inhibition and oxygen supply is lacking. Therefore, fully equipped laboratory-scale stirred tank bioreactors are hitherto required for detailed studies of new microbial systems. However, they are too spacious, laborious and expensive to be operated in larger number in parallel. Thus, the aim of this study is to present a new experimental approach to obtain dense quantitative process information by parallel use of two small-scale culture systems with online monitoring capabilities: Respiration Activity MOnitoring System (RAMOS) and the BioLector device. RESULTS: The same ‘mastermix’ (medium plus microorganisms) was distributed to the different small-scale culture systems: 1) RAMOS device; 2) 48-well microtiter plate for BioLector device; and 3) separate shake flasks or microtiter plates for offline sampling. By adjusting the same maximum oxygen transfer capacity (OTR(max)), the results from the RAMOS and BioLector online monitoring systems supplemented each other very well for all studied microbial systems (E. coli, G. oxydans, K. lactis) and culture conditions (oxygen limitation, diauxic growth, auto-induction, buffer effects). CONCLUSIONS: The parallel use of RAMOS and BioLector devices is a suitable and fast approach to gain comprehensive quantitative data about growth and production behavior of the evaluated microorganisms. These acquired data largely reduce the necessary number of experiments in laboratory-scale stirred tank bioreactors for basic process development. Thus, much more quantitative information is obtained in parallel in shorter time. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13036-015-0005-0) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4531433 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-45314332015-08-12 Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors Wewetzer, S. J. Kunze, M. Ladner, T. Luchterhand, B. Roth, S. Rahmen, N. Kloß, R. Costa e Silva, A. Regestein, L. Büchs, J. J Biol Eng Research BACKGROUND: Conventional experiments in small scale are often performed in a ‘Black Box’ fashion, analyzing only the product concentration in the final sample. Online monitoring of relevant process characteristics and parameters such as substrate limitation, product inhibition and oxygen supply is lacking. Therefore, fully equipped laboratory-scale stirred tank bioreactors are hitherto required for detailed studies of new microbial systems. However, they are too spacious, laborious and expensive to be operated in larger number in parallel. Thus, the aim of this study is to present a new experimental approach to obtain dense quantitative process information by parallel use of two small-scale culture systems with online monitoring capabilities: Respiration Activity MOnitoring System (RAMOS) and the BioLector device. RESULTS: The same ‘mastermix’ (medium plus microorganisms) was distributed to the different small-scale culture systems: 1) RAMOS device; 2) 48-well microtiter plate for BioLector device; and 3) separate shake flasks or microtiter plates for offline sampling. By adjusting the same maximum oxygen transfer capacity (OTR(max)), the results from the RAMOS and BioLector online monitoring systems supplemented each other very well for all studied microbial systems (E. coli, G. oxydans, K. lactis) and culture conditions (oxygen limitation, diauxic growth, auto-induction, buffer effects). CONCLUSIONS: The parallel use of RAMOS and BioLector devices is a suitable and fast approach to gain comprehensive quantitative data about growth and production behavior of the evaluated microorganisms. These acquired data largely reduce the necessary number of experiments in laboratory-scale stirred tank bioreactors for basic process development. Thus, much more quantitative information is obtained in parallel in shorter time. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13036-015-0005-0) contains supplementary material, which is available to authorized users. BioMed Central 2015-05-30 /pmc/articles/PMC4531433/ /pubmed/26265936 http://dx.doi.org/10.1186/s13036-015-0005-0 Text en © Wewetzer et al. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Wewetzer, S. J. Kunze, M. Ladner, T. Luchterhand, B. Roth, S. Rahmen, N. Kloß, R. Costa e Silva, A. Regestein, L. Büchs, J. Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors |
title | Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors |
title_full | Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors |
title_fullStr | Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors |
title_full_unstemmed | Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors |
title_short | Parallel use of shake flask and microtiter plate online measuring devices (RAMOS and BioLector) reduces the number of experiments in laboratory-scale stirred tank bioreactors |
title_sort | parallel use of shake flask and microtiter plate online measuring devices (ramos and biolector) reduces the number of experiments in laboratory-scale stirred tank bioreactors |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4531433/ https://www.ncbi.nlm.nih.gov/pubmed/26265936 http://dx.doi.org/10.1186/s13036-015-0005-0 |
work_keys_str_mv | AT wewetzersj paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT kunzem paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT ladnert paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT luchterhandb paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT roths paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT rahmenn paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT kloßr paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT costaesilvaa paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT regesteinl paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors AT buchsj paralleluseofshakeflaskandmicrotiterplateonlinemeasuringdevicesramosandbiolectorreducesthenumberofexperimentsinlaboratoryscalestirredtankbioreactors |