Coflobiopro: Continuous flow biocatalytic process for production of value-added chemicals using magnetic nanoparticle immobilized enzyme microreactors
Lead Research Organisation:
UNIVERSITY COLLEGE LONDON
Department Name: Biochemical Engineering
Abstract
The Coflobiopro project will establish an optimally functioning continuous enzyme cascade with pure glycerol as substrate to produce acetoin and 2,3-butanediol(2,3BD) as the major products. 2,3-BD has an extremely high industrial value with potential application in the sectors of aviation fuel, rubber, pharmaceuticals, methyl-ethyl ketone (fuel additive), foodstuff(diacetyl) and acetoin production (volatile food additive). The current global market values of the major downstream products from 2,3-BD is reported to be $43 billion per year with the value of acetoin reported to be approximately $30000 per ton. The project will utilize currently prevailing technological advancements in a catalytic flow through system packed in a single microreactor platform. Effective research strategies have been put forth towards generation of 2, 3 butanediol (2,3-BD) by bioengineered microbial species from renewable substrates but with limited success rate since the terminal product is low in volume. To counter this, we have devised a synthetic metabolic pathway mediated by magnetic nanoparticle (MNP) immobilized biocatalysts in a packed bed enzymatic microreactor. Initially, we will ascertain the optimal conditions for product outputs for each of the sequential reaction steps in the study. The individual reactions catalyzed by the MNP immobilized enzymes alditol oxidase, dihydroxy acid dehydratase, a-acetolactate synthase, a-acetolactate decarboxylase and butanediol dehydrogenase in individual reactors will determine the conditions that lends optimal output of the resultant products. Finally, the optimized proportions of the MNP-immobilised enzymes will be loaded in a single reactor and the reactions will be performed under optimized conditions in continuous mode. The novelty of Coflobiopro is that it incorporates optimally functioning MNP immobilized enzymes aligned in a sequential manner in a microreactor for generation of
value-added biofuels in a continuous biocatalytic cascade.
value-added biofuels in a continuous biocatalytic cascade.
Organisations
Publications
Jiang W
(2024)
Inactivation of hydrogenase-3 leads to enhancement of 1,3-propanediol and 2,3-butanediol production by Klebsiella pneumoniae.
in Enzyme and microbial technology
| Description | 1. Successful Construction of an in vitro Enzyme Cascade This project successfully developed a enzymatic reaction cascade within a packed-bed immobilized enzyme microreactor employing nanoparticle-immobilized enzymes to efficiently convert glycerol into value-added chemicals. The enzymatic cascade involved the sequential action of five enzymes (i.e. dihydroxy-acid dehydratase , alditol oxidase, a-acetolactate synthase, a-acetolactate decarboxylase, and butanediol dehydrogenase) optimized for acetoin and 2,3-butanediol (2,3-BD) production. This reaction sequence enabled high catalytic efficiency, enhanced substrate utilization, and improved reaction specificity, thereby increasing overall product yield. Furthermore, the microfluidic reactor design ensured precise control over reaction conditions, minimizing issues related to diffusion constraints and pH fluctuations. This advancement significantly enhanced the reproducibility and reliability of the process. 2. Process Intensification and Enhanced Biocatalyst Performance One of the key advantages of this approach was the ability to achieve continuous operation under constant conditions. Notably, the use of magnetic nanoparticles for enzyme immobilization resulted in higher catalytic activity, increased stability, and enhanced resistance to thermal and pH variations compared to using free enzymes in solution. The immobilized enzymes exhibited greater structural integrity and longer functional lifespan, mitigating issues of enzyme degradation typically encountered in free enzyme systems. This enhanced durability is particularly significant for industrial applications, as it reduces the need for frequent enzyme replenishment, thereby lowering operational costs and improving economic feasibility. Additionally, the high surface-to-volume ratio of the microfluidic reactor facilitated efficient mass transfer, ensuring consistent substrate conversion and reducing reaction times. 3. Optimized Reaction Parameters and Yield Enhancement The project established optimal conditions for maximizing product yield, including substrate concentration, cofactor availability, enzyme loading, and reaction time. The controlled supply of NADH/NAD+ was particularly critical for efficient conversion of acetoin to 2,3-BD, as the activity of butanediol dehydrogenase is directly influenced by the redox balance. Through extensive process optimization the microfludic reactor system achieved: • High selectivity towards 2,3-BD while minimizing unwanted side products. • Improved reaction kinetics and accelerated conversion rates compared to free enzyme systems. • Scalability potential, demonstrating feasibility for industrial-scale implementation. |
| Exploitation Route | One of the most significant outcomes of this project is its translational potential-moving beyond academic advancements to industrial applications. By demonstrating the feasibility of continuous bioproduction at a microfluidic scale, this project paves the way for scaling up enzymatic bioprocesses for large-scale chemical synthesis. Potential applications include: • Green production of industrial solvents (acetoin) and biodegradable polymers (2,3-BD). • High-efficiency biochemical pathways for pharmaceutical precursors. • Sustainable alternatives to fossil-derived fuels and chemicals, reducing carbon emissions and reliance on non-renewable resources. This research also sets the stage for further advancements in enzyme immobilization technology, potentially leading to: • Next-generation biocatalytic microreactors, capable of multi-step biochemical transformations. • Customizable multi-enzyme systems, tailored for diverse industrial needs. • Automated, AI-driven process optimization, enhancing efficiency and adaptability. |
| Sectors | Chemicals Manufacturing including Industrial Biotechology |