Our team from Ghent University has released a Mini-Review presenting an overview of the application of β-glucan phosphorylases in carbohydrate synthesis. Derived carbohydrates (β-glucans) such as cellodextrins can be used as ingredients and additives in the food, feed or cosmetic industry. There is no doubt that the relevance of this enzyme class is increasing for these industrial sectors and will warrant spending more efforts on enzyme engineering, as was also shown in the CARBAFIN project.
Have a look on the publication by following the link:
CARBAFIN has released a review about cellodextrin phosphorylase and its suitability for a bottom-up production of cellulose derivatives. So far, these derivatives are produced top-down from lignocellulosic substrates. However, for some product classes a more controllable and “property-tunable” approach would be clearly preferred (eg hydrogels). The publication explores the characteristics of the enzyme and the advantages of an enzymatic production of soluble cello-oligosaccharide.
The optimization of chemical reactions always implies to know the underlaying kinetics like the back of one’s hand. This becomes even more important if a reaction is going to be applied for commercial-scale production! Sucrose phosphorylase, the central enzyme of CARBAFIN, was characterized in terms of kinetics related to the donor substrate: for the glycosylation of glycerol, the effect of donor substrates, in particular sucrose and glucose-1-phosphate was explored.
The results, published by CARBAFIN partner Graz University of Technology, give an explanation for the kinetic behavior of sucrose phosphorylase and show that type and concentration of the glucosyl donor have a significant impact on enzyme selectivity for product formation.
Sugar industry is facing a crisis since several years. The sugar quota has fallen and caused a significant descrease in prices. The European project CARBAFIN is about the development of a radically new value chain for the utilization of sucrose from sugar beet biomass. Functional glycosides (eg for prebiotics or cosmetics) or platform chemicals (eg for bioplastics, biopolymers or biofuels) are important examples for products of the new technology and open new and economically relevant perspectives to several industry sectors.
The central aspect for the conversion of sugar (sucrose) is the use of glycoside phosphorylases, highly active and robust enzymes for linking sugar molecules to another compound. In the frame of CARBAFIN this glycosylation process was optimized within cell factories and linked to an efficient downstream processing.
What does that mean to the industrial sectors of food/feed, cosmetics or also chemistry?
The webinar demonstrates how CARBAFIN is going to contribute to a bio-based re-industrialization in Europe by glycosylation processes based on sucrose with fructose valorization in parallel.
We cordially invite exploiters and potential technology users from industry as well as interested scientists to join our virtual CARBAFIN event and to discuss potential application fields and perspectives!
Welcome address (Prof. Dr. Bernd Nidetzky; Coordinator of CARBAFIN, CSO acib GmbH, Professor at Institute of Biotechnology and Biochemical Engineering, Graz University of Technology)
CARBAFIN – a phosphorylase-based glycosylation technology
Introduction to the project: CARBAFIN at a glance (Dr. Christiane Luley; Project manager of CARBAFIN, acib GmbH)
Process integration strategies (DI Andreas Kruschitz; PhD student of CARBAFIN, acib GmbH)
When it comes to application – experiences from industry
Single phosphorylase glycosylation technology for Glycoin® production at bitop
Multi-step phosphorylase glycosylation technology in bulk industry at Pfeifer & Langen
Making use of fructose (waste) streams for the production of 5-HMF at AVA Biochem
The potential of generating new value chains
Potential of CARBAFIN technology in other industry sectors (Dr. Kai Baldenius; External Advisor of CARBAFIN, Consultant)
CARBAFIN has started into the final year of project duration and is approaching a couple of events. You are interested in the technologies, the results and the outcomes of the project and you want to know, how you could benefit from our findings? Don’t miss our webinar in spring, our public event in summer or our Stakeholder conference session at EFIB 2021 in Vienna. Find below a list of events, where CARBAFIN is going to be presented this year. We are looking forward to meet you soon. Stay tuned!
Stakeholder Webinar: The CARBAFIN glycosylation technology – generating new value chains in a novel integrated biocatalytic production technology and the implementation in industry (April 2021, registration starts mid of February)
Public Event: Sweet breakfast – Find out about the biotechnology behind novel food and cosmetic ingredients. Why is it important to identify radically new value chains for the utilization of biomass? (Summer 2021)
CARBAFIN session at EFIB 2021: The CARBAFIN glycosylation technology and its future perspectives – what comes next? (5th – 7th October 2021)
CARBAFIN has finished its third year of project duration and sums up the intermediate status of project results and activities in the recent newsletter (released on 7th December 2020). Enjoy reading!
Before entering the fourth and final year of CARBAFIN, the whole team wishes you a joyful and merry Christmas! All the best for 2021 and don’t forget about the sweet side of life in these challenging times 🙂
The CARBAFIN team of UGent reported a successful application of engineered enzymes for the production of cellotriose! Cellotriose belongs to the group of cellodextrins, compounds that have the potential to act as prebiotics. Currently, cellodextrins are synthesized from cellulose by the chemical procedures that result in a complex mixture of oligosaccharides.
The idea within CARBAFIN is to produce cellotriose, believed to be the most potent component in the mixture, by an enzymatic bottom-up approach that would enable its defined synthesis. Cellotriose is composed of 3 glucose-units. Two bacterial strains (Cellulomonas uda and Clostridium cellulosi) provide the most promising enzyme candidates, which can be applied in a one-pot reaction. In the first step, enzyme number 1 (cellobiose phosphorylase, CBP) from Cellulomonasuda assembles two elements to produce cellobiose. Then, enzyme number 2 (cellodextrin phosphorylase, CDP) from Clostridium cellulosi elongates cellobiose by other glucose units to produce the three-piece compound, cellotriose but also larger cello-oligosaccharides.
In the recent paper, a special focus is given to the CBP enzyme and its improved variant that can synthesize cellotriose on its own. It represents a crucial step in enabling further research and commercial exploitation of cellotriose.
Cellodextrins are functional carbohydrates that are expected to be promising water-soluble dietary fibres. They can act as a prebiotic ingredient and, therefore, have a positive effect on health: Prebiotics are stimulating the immune system, regulating the gastrointestinal function or preventing from carcinogenesis for example. An important criterium for prebiotic components is that they are not digested by humans. Cello-oligosaccharides (COS), also known as cellodextrin meet this requirement and are, therefore, one of the main target products of the CARBAFIN project.
The recent publication explains the enzymatic production of COS at larger scale in order to bring the technology that has been proven previously, to the next level and to open the door towards industrial application. The CARBAFIN researchers from acib and TU Graz together with researchers from BOKU demonstrate the soluble COS production to approximately 100 gram per litre in a 20 mL reaction volume. The enzymatic reaction turned out to be very efficient and to provide a product with a very satisfying yield and purity from simple and expedient substrates. The stimulation of growth among probiotic bacteria was also shown.
Cellobiose, the simplest cellodextrin, is one of CARBAFIN’s target products, derived from sugar (glucose) molecules. The biocatalytic production process is divided into two steps:
Enzyme No. 1 (sucrose phosphorylase) produces an activated glucose component from sucrose, namely glucose-1-phosphate.
Enzyme No. 2 (cellobiose phosphorylase) uses the glucose-1-phosphate from the prior reaction and glucose for producing cellobiose.
For a cascade reaction like this, a combined process within one cell would be beneficial. But convincing the cell to do so, comes along with a huge effort of research. The CARBAFIN team was successful in designing an appropriate tool (plasmid) that allows the co-production of the two enzymes in perfect relation to each other. The obtained process and derived knowledge can be easily transferred to other cascadic reactions like this and, therefore, is of broad interest across the whole field of biocatalysis.