Azzoni, Sindelia Freitas

Bussamra, Bianca Consorti

da Costa, Aline Carvalho

Gomes, Joana Castro

Mussatto, Solange I.

Ottens, Marcel

van der Wielen, Luuk A. M.

Online Research Database In Technology

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Downloaded from orbit.dtu.dk on: Dec 20, 2017From bench-scale to high throughput platform: Phase diagrams determination andsolute partition analysis in aqueous two phase systemsBussamra, Bianca Consorti; Azzoni, Sindelia Freitas; Mussatto, Solange I.; da Costa, Aline Carvalho;van der Wielen, Luuk A. M.; Gomes,  Joana Castro; Ottens, MarcelPublication date:2017Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Bussamra, B. C., Azzoni, S. F., Mussatto, S. I., da Costa, A. C., van der Wielen, L. A. M., Gomes, . J. C., &Ottens, M. (2017). From bench-scale to high throughput platform: Phase diagrams determination and solutepartition analysis in aqueous two phase systems. Abstract from Anything but conventional chromatography,Lisbon, Portugal.ABC2 – Anything but Conventional Chromatography  -55- OP17 - From bench-scale to high throughput platform: Phase diagrams determination and solute partition analysis in aqueous two phase systems  Bianca Consorti Bussamra1,2, Sindelia Freitas Azzoni2, Solange I. Mussatto3, Aline Carvalho da Costa2, Luuk van der Wielen1, Joana Castro Gomes1, Marcel Ottens1  1Delft University of Technology. Van der Maasweg 9, 2629HZ. Delft, The Netherlands.  2University of Campinas. Av. Albert Einstein, 500. Post Code: 6066. Campinas, Brazil.  3Technical University of Denmark. Kemitorvet, Building 220. 2800, Kongens Lyngby, Denmark. b.consortibussamra@tudelft.nl  Sugars derived from lignocellulosic materials are the main raw material in bio-based processes aiming to produce renewable fuels and chemicals. However, inhibition of enzymes by reaction products (cellobiose and glucose) is one of the major drawbacks of enzymatic hydrolysis of lignocellulosic materials. This effect is even more pronounced in hydrolysis containing high solid content (15-20% or higher water-insoluble solids – WIS), which is desired in order to result in higher total reducing sugar concentration in hydrolysates and reduce water usage1. Aqueous Two-Phase Systems (ATPSs) are used in a wide range of processes, including analytical applications, product recovery and environmental remediation. In the case of product recovery, ATPS enables an integrated process in which the bioconversion occurs at the same time that the products are being removed from the system. This approach, called extractive bioconversion, contributes the most to reactions in which the products are inhibitors 2. To the best of our knowledge, ATPS has not been used as an extractive environment to relieve product inhibition in biomass conversion. Although works using high throughput platforms have been largely reported in literature3 4 5 6 7, there is a lack of technologies that address high throughput screening for separation of sugars and proteins in aqueous two-phase systems. This paper will present the methodology development and its transference from bench-scale to a high throughput platform for screening and selection of ATPS. Determination of phase diagrams, partition coefficients of enzymes and proteins, enzymatic activities, as well as a strategy designed to optimize the system parameters (pH, temperature and salt concentration) will be shown. Therefore, this automated and miniaturized experimental setup will be used to develop a new process for sugarcane bagasse hydrolysis based on aqueous two-phase system. The purpose of this technique is to remove the product is situ, as long as it is produced. As a consequence of product removal, enzymes tend to maintain their maximum activity8. ABC2 – Anything but Conventional Chromatography  -56-  References [1] Modenbach, A. A. & Nokes, S. E. Enzymatic hydrolysis of biomass at high-solids loadings - A review. Biomass and Bioenergy 56, 526–544 (2013); [2] Hatti-Kaul, R. Aqueous two-Phase Systems. Methods and Protocols. 19, (Human Press, 2000); [3] Bensch, M. et al. High throughput screening techniques in downstream processing: Preparation, characterization and optimization of aqueous two-phase systems. Chem. Eng. Sci. 62, 2011–2021 (2007); [4] Amrhein, S. et al. Characterization of aqueous two phase systems by combining lab-on-a-chip technology with robotic liquid handling stations. J. Chromatogr. A 1367, 68–77 (2014); [5] Chundawat, S. P. S. et al. High-throughput microplate technique for enzymatic hydrolysis of lignocellulosic biomass. Biotechnol. Bioeng. 99, 1281–1294 (2008); [6] Nfor, B. K. et al. High-throughput isotherm determination and thermodynamic modeling of protein adsorption on mixed mode adsorbents. J. Chromatogr. A 1217, 6829–6850 (2010); [7] Zimmermann, S. et al. High-throughput downstream process development for cell-based products using aqueous two-phase systems. J. Chromatogr. A 1464, 1–11 (2016); [8] Yang, J. et al. Three-stage enzymatic hydrolysis of steam-exploded corn stover at high substrate concentration. Bioresour. Technol. 102, 4905–4908 (2011).  Keywords: Aqueous two-phase systems; high throughput experimentation; product inhibition; separation of sugar and enzymes.              Book of Abstracts      ABC2 – Anything but Conventional Chromatography  -1-    Anything but Conventional Chromatography    22nd – 24th November 2017  

From bench-scale to high throughput platform: Phase diagrams determination and solute partition analysis in aqueous two phase systems

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From bench-scale to high throughput platform: Phase diagrams determination and solute partition analysis in aqueous two phase systems

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