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12. An adsorptive bioprocess for production and recovery of resveratrol with Corynebacterium glutamicum

Author

Braga, Adelaide, Silva, Marcelo, Oliveira, Joana Filipa Azevedo, Silva, Rita M., Ferreira, Patrícia, Ottens, Marcel, Rocha, I., Faria, Nuno, and Universidade do Minho

Subjects
Corynebacterium glutamicum, In-situ product removal (ISPR), Science & Technology, Direct adsorption, Resveratrol, Amberlite XAD7-HP, food and beverages, External adsorption
Abstract

BACKGROUND The growing interest in polyphenols has led to the design of industrial-scale processes able to produce them by fermentation and recover them in a more sustainable way. The goal of this work is to present two integrated approaches for the recovery of resveratrol, obtained through fermentation. The production of resveratrol using Corynebacterium glutamicum and its continuous removal using a hydrophobic resin is described. Batch production is compared with in situ product removal, where Amberlite XAD-7HP is either directly added to the medium (direct adsorption) or is present in an external column (external adsorption). RESULTS For both adsorption strategies tested, the amount of extracellular resveratrol increased from 75% to at least 90% of the total amount produced. However, lower total resveratrol concentrations were attained 3.6 and 2.2mg L-1, for the external and direct contact strategies, respectively, versus 5.3mg L-1 for batch experiments. CONCLUSIONS The proposed in situ removal strategies demonstrated the potential of increasing the excretion of resveratrol produced intracellularly. These process configurations may not only lead to a simpler downstream process design, but also to the avoidance of potential problems with the toxicity of polyphenols to the cells, especially when larger titers are obtained. © 2017 Society of Chemical Industry, We would like to thank the European Union Framework Program 7 ‘BacHBerry’ (www.bachberry.eu), Project No. FP7- 613793 for financial support, the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit, COMPETE 2020 (POCI-01- 0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145- FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte., info:eu-repo/semantics/publishedVersion

Published
2018

26. Multi-dimensional fractionation and characterization of crude protein mixtures: Toward establishment of a database of protein purification process development parameters

Author

Nfor, Beckley K., primary, Ahamed, Tangir, additional, Pinkse, Martijn W.H., additional, van der Wielen, Luuk A.M., additional, Verhaert, Peter D.E.M., additional, van Dedem, Gijs W.K., additional, Eppink, Michel H.M., additional, van de Sandt, Emile J.A.X., additional, and Ottens, Marcel, additional

Published
2012
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29. Precipitation of porcine insulin with carbon dioxide

Author

Tashima, Alexandre Keiji, primary, Ottens, Marcel, additional, Van der Wielen, Luuk A.M., additional, Cintra, Dennys E., additional, Pauli, José R., additional, Filho, Pedro de Alcântara Pessôa, additional, and Miranda, Everson Alves, additional

Published
2009
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38. Experimental characterization and prediction of Escherichia coli host cell proteome retention during preparative chromatography.

Author

Disela R, Neijenhuis T, Le Bussy O, Geldhof G, Klijn M, Pabst M, and Ottens M

Abstract

Purification of recombinantly produced biopharmaceuticals involves removal of host cell material, such as host cell proteins (HCPs). For lysates of the common expression host Escherichia coli (E. coli) over 1500 unique proteins can be identified. Currently, understanding the behavior of individual HCPs for purification operations, such as preparative chromatography, is limited. Therefore, we aim to elucidate the elution behavior of individual HCPs from E. coli strain BLR(DE3) during chromatography. Understanding this complex mixture and knowing the chromatographic behavior of each individual HCP improves the ability for rational purification process design. Specifically, linear gradient experiments were performed using ion exchange (IEX) and hydrophobic interaction chromatography, coupled with mass spectrometry-based proteomics to map the retention of individual HCPs. We combined knowledge of protein location, function, and interaction available in literature to identify trends in elution behavior. Additionally, quantitative structure-property relationship models were trained relating the protein 3D structure to elution behavior during IEX. For the complete data set a model with a cross-validated R 2 of 0.55 was constructed, that could be improved to a R 2 of 0.70 by considering only monomeric proteins. Ultimately this study is a significant step toward greater process understanding., (© 2024 The Author(s). Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

Published
2024
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39. Using artificial neural networks to accelerate flowsheet optimization for downstream process development.

Author

Keulen D, Hagen EV, Geldhof G, Le Bussy O, Pabst M, and Ottens M

Subjects
Chromatography methods, Neural Networks, Computer
Abstract

An optimal purification process for biopharmaceutical products is important to meet strict safety regulations, and for economic benefits. To find the global optimum, it is desirable to screen the overall design space. Advanced model-based approaches enable to screen a broad range of the design-space, in contrast to traditional statistical or heuristic-based approaches. Though, chromatographic mechanistic modeling (MM), one of the advanced model-based approaches, can be speed-limiting for flowsheet optimization, which evaluates every purification possibility (e.g., type and order of purification techniques, and their operating conditions). Therefore, we propose to use artificial neural networks (ANNs) during global optimization to select the most optimal flowsheets. So, the number of flowsheets for final local optimization is reduced and consequently the overall optimization time. Employing ANNs during global optimization proved to reduce the number of flowsheets from 15 to only 3. From these three, one flowsheet was optimized locally and similar final results were found when using the global outcome of either the ANN or MM as starting condition. Moreover, the overall flowsheet optimization time was reduced by 50% when using ANNs during global optimization. This approach accelerates the early purification process design; moreover, it is generic, flexible, and regardless of sample material's type., (© 2023 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

Published
2024
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