Team:UCL/Science/Bioprocessing

From 2014.igem.org

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<b>Fermentation:</b> through adequate bioreactor dimensions and operating parameters, optimized E. coli (expressing BsDyP) cultures are carried out in batch mode. The target yield > 90% is necessary to ensure enough biomass is loaded onto the trays in the subsequent unit.  
<b>Fermentation:</b> through adequate bioreactor dimensions and operating parameters, optimized E. coli (expressing BsDyP) cultures are carried out in batch mode. The target yield > 90% is necessary to ensure enough biomass is loaded onto the trays in the subsequent unit.  
<br><b>Separation:</b>since the enzyme is expressed intra-cellularly and the immobilisation module is equipped with a rough filter, separation is not a necessity if the fermenter is integrated in the tertiary treatment process. However, the E. coli would usually be separated from the broth if produced and supplied by a specialist company (bioreactor not integrated).
<br><b>Separation:</b>since the enzyme is expressed intra-cellularly and the immobilisation module is equipped with a rough filter, separation is not a necessity if the fermenter is integrated in the tertiary treatment process. However, the E. coli would usually be separated from the broth if produced and supplied by a specialist company (bioreactor not integrated).
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<br><b>Module operation:</b> see cross section of a single system. Continuous flow system with flow rates and residence times based on mass transfer kinetics, specific to E. Coli
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<br><b>Module operation:</b>Azodye degradation takes place here. E. coli from the upstream section is immobilized by chemical means (i.e. alignate) or by biofilm formation (other methods can also be considered). These are formed on removable trays, which are stacked within the unit. The azodye effluent stream from secondary treatment, enriched with a carbon source, is then introduced into the unit from the top and flowed through the module continuously until an acceptable decolourization is achieved – glass viewing points and sampling ports for on-line analysis are the two methods to quantify this. The first module is designed to remediate bulk azodyes whereas the second serves as a polishing step. Air inlets allow for the aeration of trays, depending on the mode of operation – aerobic vs. anaerobic. Filter meshes installed on each tray act as an initial separation process, which could also serve as a biomass concentration mechanism post-fermentation.
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<br> - Module 1 designed to capture the bulk of the azodyes, module 2 is a polishing step
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<br><b>Further processing:</b>Additional downstream purification units such as size exclusion chromatography or liquid-liquid extraction can be incorporated subsequently in order to harvest byproducts of interest, based on their commercial value. Further economic considerations need to be carried out in order to determine the feasibility these steps to be integrated within the textile plant. Thus, a simple concentration step involving a mixer settler unit already used in the secondary treatment process could be utilized to reduce process volumes. These can then be shipped to a purification company for further processing.
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<br> - Both anaerobic and aerobic reactions take place at the same time in both the modules, design based on gas supply (nitrogen vs. oxygen)
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<br> - Cleaning operation using biodegradable chemical at high flow rate (from holding tank 2)
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<br> - Continuous recycle system for maximal active and diffusive uptake.
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<br> - Filter modules- exploring the use of disposable low cost agricultural waste for filtration
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<br> - Further processing- based on the commercial value of the breakdown products, investments could be made into higher-tier technology such as chromatography columns to separate the breakdown products individually.
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<p>This versatile and simple process offers a wide range of future developments into various chemical producing sectors. It would be possible to use this technology in parallel with different industries as a form of platform technology using different synthetic biology anchors, in order to detoxify various effluent polluting chemicals.</p>
 
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<div class="textTitle"><h4>Our Design Process</h4></div>
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<div class="textTitle"><h4>Future trends</h4></div>
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<p>This <b>versatile</b> and simple process offers a wide range of future developments into various chemical producing sectors. It would be possible to use this technology in parallel with different industries as a form of platform technology using different synthetic biology anchors, in order to detoxify various effluent polluting chemicals.</p><br>
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<iframe style="padding:1%; border:0.5% #000;" src="//www.youtube.com/embed/6UNpqpMS5vA" allowfullscreen="" align="right" frameborder="0" height="380" width="600"></iframe></div>
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<p>We will use rapid polymer prototyping techniques to generate microfluidic chips that will allow us to test our reaction and aid in the construction of a realistic bioprocess, which can be successfully scaled-up for industrial use. As we optimise and change our bioprocess, we can also quickly design new microfluidic chips that can mimic its development on a micro-scale. For example, it is our goal to integrate multiple downstream steps, such as chromatography, in order to isolate potential useful products. Demonstrating this in a microfluidic system is less time-consuming and far more cost effective than doing so at a larger scale.</p><br>
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<p>For our microfluidic bioreactor, we will be using a magnetic free floating bar as our mixing system. This is an effective method of mixing at a microfluidic scale, as demonstrated in the video on the right. This video is of a microfluidic chemostat bioreactor designed by Davies et al. 2014 UCL, using a free-floating bar to mix two dyes.</p><br>
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<img src="https://static.igem.org/mediawiki/2014/thumb/b/b8/AutoCAD_Device.png/800px-AutoCAD_Device.png" width="32%">
<img src="https://static.igem.org/mediawiki/2014/thumb/b/b8/AutoCAD_Device.png/800px-AutoCAD_Device.png" width="32%">

Revision as of 12:26, 17 October 2014

Goodbye Azodye UCL iGEM 2014

Sustainable Bioprocessing

Design of immobilization unit


After the fermentation stage, the E. coli biomass is dispersed in a liquor also containing various byproducts. A concentration step could be beneficial to reduce volumes in the next stage. However, capital costs of such unit operations would not be attractive to potential dyeing companies deciding to acquire the entire system. The subsequent modules are equipped to handle large volumes and operate in continuous-flow mode with intermittent discharges. By controlling residence time and operating flow rates, it will be possible to achieve a cell recovery deemed efficient. These will be then immobilized onto the surface of the plates within the modules. There exists a wide range of immobilization strategies used for biological wastewater treatment and this is what gives the unit its modular character. By supporting a number of immobilization methods without changing the hardware, the module allows for the enzymatic breakdown of a wide range of recalcitrant chemicals that might be financially and environmentally costly to treat using conventional methods.



Top view of the module with Azodye feed pipe (red) and aeration inlets for the plates (green).

Conclusions

This system is designed.
References


- Ilgi Karapinar Kapdan, F. K. (2002). Simultaneous Degradation and Adsorption of Textile Dyestuff in an Activated Sludge Unit. Process Biochemistry, 973-981.
- Michael Winn, J. F. (2012). Biofilms and Their Engineered Counterparts: A new generation of immobilised biocatalysts. Catalysis Science and Technology, 1544-1547.
- Rajbir Singh, D. P. (2006). Biofilms: Implications in Bioremediation. Trends in Microbiology, 389-396.
- Sethi Sonia, Subhum, Malviya M. Mukesh et al.(2012). Biodecolorization of Azodye by Microbial isolates from textile effluent and sludge. Universal Journal of Environmental research and technology. Vol. 2, issue 6:582-590.


Future trends

This versatile and simple process offers a wide range of future developments into various chemical producing sectors. It would be possible to use this technology in parallel with different industries as a form of platform technology using different synthetic biology anchors, in order to detoxify various effluent polluting chemicals.



Above are some examples of the microfluidics devices developed by our team for use in the lab at the UCL ACBE. The devices are initially designed using AutoCAD (2D and 3D computer-aided design software), once the designs are finalised they can be 3D-printed using the facilities provided by the UCL Institute of Making and UCL ACBE; allowing our bioprocess and laboratory team to experiment and improve designs.


An example of one of our microfluidic devices designed on AutoCAD can be downloaded here. This device utilises the basic concept of mixing the cells and dyes, producing a single output stream; much alike to the bioprocessing concept. During the course of designing the microfluidic device, several key considerations must be taken into account: ability to withstand high pressure without leakage; materials of construction to be inert and transparent; size constraints of inlet and outlet piping; ability to accurately 3D-print the device.


Why Bioprocessing?

Bioprocess engineering is a conglomerate of fields and is extensively employed to optimize a variety of production processes. In order to cope with market forces, industries for example the pharmaceutical, have had to considerably improve their bioprocessing tools and techniques. As a result a range of novel process alternatives have been developed to harness product-specific properties, each bearing benefits, disadvantages and costs. While these can be used to drive financial returns, biological processing is becoming a gateway to eco-friendly alternatives for the treatment of recalcitrant wastewater such as industrial effluents. By providing more flexibility in supporting efficient degradation of toxic compounds and having lower operating costs, the biological treatment process brings forward key advantages over it's traditional counterpart.
A typical bioprocess involves the fermentation of a stock culture (e.g. E. coli) at a small scale which is subsequently scaled up to suitable production capacities. The products from the fermentative stages are consequently separated and purified using a variety of unit operations designed to exploit the orthogonal properties of desired products. These can then be formulated into their ultimate delivery form.




Flowsheet with unit operations for a typical bioprocess

The design of a successful bioprocess requires careful analysis of the many factors that impact choice of design parameters and process variables. It is crucial to consider the cost of the process at each stage to assess it's large scale feasibility.

Let's look at an example bioprocess
1. Upstream: Production bioreactor preceded by small-scale seed fermenters
2. Downstream: constitutes of three main stages
- Recovery relates to primary unit operations i.e. centrifugation and filtrations. The main goal is to concentrate the desired compound within the process stream by reducing volumes and removing fermentation byproducts.
- Purification involves unit operations such as chromatography, crystallization and ultrafiltration. The final stages are necessary to ensure purity requirements are met.
- Formulationinvolves the integrating of the product into the target delivery route followed by packaging and storage.

Contact Us

University College London
Gower Street - London
WC1E 6BT
Biochemical Engineering Department
Phone: +44 (0)20 7679 2000
Email: ucligem2014@gmail.com

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