Discover the future of chitosans!
Successful Nano3Bio dissemination session at EuroCarb 2017 in Barcelona
During the 19th European Carbohydrate Symposium EuroCarb 2017 in July 2017, organised in Barcelona by Nano3Bio partner Prof. Toni Planas from Ramon Llull University, one of the parallel afternoon sessions on Monday was fully devoted to our European research project Nano3Bio. The Nano3Bio dissemination session started with a key note lecture by the coordinator of the project, Prof. Bruno Moerschbacher from the University of Münster in Germany, who gave an introduction and overview of the whole project, explaining why it aims to develop biotechnological ways to produce well-defined chitosan oligomers and polymers with non-random patterns of acetylation. Moerschbacher described the two parallel approaches the project pursues towards this goal, namely the in vitro bio-refinery approach and the in vivo cell-factory approach, and highlighted the successes so far reached. Prof. Marjan de Mey from the University of Ghent in Belgium then explained how metabolic engineering can improve yields and purity of the monoclonal chitosan oligomers produced in bacteria; Prof. Francisco Goycoolea, now at the University of Leeds in the UK, described different nanoformulations of chitosans and how they can support gene delivery and novel antimicrobial strategies based on quorum quenching. Dr. Christian Gorzelanny from the University of Mannheim/Heidelberg in Germany showed how chitosan-based nanocapsules can be targeted to tumour vessels, delivering potential cargo drugs to their target site. Finally, Dr. Stefan Cord-Landwehr from the Münster group and Hugo Aragunde from the Barcelona group had been selected to present their posters on quantitative mass spectrometric sequencing of partially acetylated chitosan oligomers and on a screening strategy to identify optimized versions of a bacterial chitin deacetylase from a mutein library, respectively, in short talks. The Nano3Bio session was well-visited and included very good discussions which extended into the poster session afterwards, approached by a good number of colleagues from around the world who were quite impressed by the project results and interested in future collaborations - an excellent result of the Nano3Bio dissemination meeting.
The biannual EuroCarb meeting series began almost forty years ago with a small meeting devoted to carbohydrate chemistry, and it has grown ever since. This year’s meeting attracted almost 700 participants from all over the world, and the organisers successfully managed to develop this meeting into one that bridges glyco-chemistry and glyco-biology, making it an ideal opportunity to disseminate the results of Nano3Bio.
Nano3Bio's major achievements so far
Biotechnology provides novel chitosans
Chitosans are a promising class of biopolymers with many potential applications, but the chitosans commercially available today often do not fulfil the requirements for sensitive markets such as pharmaceutics or cosmetics. One problem lies in batch-to-batch differences typically observed with chitosans produced from shrimp shell chitin, a waste by-product of shrimp peeling factories. Also, the animal origin of these chitosans is sometimes considered as problematic. Therefore, the Nano3Bio consortium aims to produce well defined chitosans with known structures and functionalities through biotechnological approaches. The tools required for this approach come from nature itself, namely enzymes such as chitin synthase which produce chitin from small sugar molecules, and chitin deacetylases which convert chitin into chitosans. Work at the University of Münster, the co-ordinating partner of the Nano3Bio project, has now identified a number of genes from different organisms - bacteria, fungi, viruses, algae - that appear to code for chitin deacetylases. These genes were used to drive the biotechnological production of the enzymes they encode. The recombinant enzymes were then characterised and used for the biotechnological conversion of chitin into chitosan. Interestingly, the chitosans obtained differ in their fine structure from all currently available chitosans which invariably are produced from chitin using chemical methods. Clearly, the chemical process yields “simple” chitosans which differ from the more complex and more varied chitosans found in nature. Ongoing work in the Nano3Bio project aims to test the material properties as well as the biological activities of these novel, “third generation” chitosans.
(Nano3Bio consortium partner in charge: University of Münster)
Learn more about this Nano3Bio achievement from project coordinator Bruno Moerschbacher in the short video below:
First detailed life cycle assessment of chitosan production
The Nano3bio project has carried out the first detailed life cycle assessment (LCA) of chitosan production from crustacean shells, using data from two manufacturers in India and Germany. LCA assess the overall environmental impacts caused by a system of production, use, and disposal processes necessary to provide a specific product. The LCA study within Nano3Bio includes the production and processing of all involved raw materials (crustacean shells as a by-product from fisheries), production of materials and energy carriers (chemicals, fuels, electricity) used in the manufacturing process, and the disposal of waste generated in the process (solid waste and wastewater). Knowledge gained from this LCA will be used by the manufacturers to define strategies to reduce their environmental impacts, both in their directly controlled activities as well as in their supply chain. The results from this study will be submitted to the European LCA database ELCD, where they will be publicly available. Implementing LCA during research activities is an important approach within the Nano3Bio project in order to contribute to an overall sustainable development regarding the usage of raw organic materials.
(Nano3Bio consortium partner in charge: 2.0 LCA Consultant APS)
- Article 'Life cycle assessment of chitosan production in India and Europe‘ in The International Journal of Life Cycle Assessment (March 2017)
- Introduction on life cycle assessment within Nano3Bio, short video below:
Internalisation of chitosan nanocapsules into human cells
Drug administration to patients is frequently associated with adverse effects. This is especially relevant for therapies of cancer with chemotherapeutics leading to reduced life quality of the patient. Adverse effects are mostly related to the inefficient deliverance of drugs to the tumour and thus to systemic dissemination affecting the whole body. In the past, various strategies have been developed to improve tumour targeting and thus to minimize adverse effect. One of the most promising approaches is the encapsulation of therapeutics into particularly small sized carriers. The development of such carriers has been intensively followed in the last decades. Today, novel chitosan-based nanocapsules represent an achievement of the Nano3Bio consortium. The chitosan applied in this context is a fully degradable biopolymer preventing the accumulation of the capsules in the human body. The efficient uptake of the chitosan-nanocapsules into certain cells due to their unique physicochemical properties envisions an improved targeting of tumours and thus a reduction of adverse effects during cancer therapies. The University of Heidelberg (Germany) mainly carried out the research and development activities related to chitosan nanocapsules.
(Nano3Bio consortium partner in charge: University of Heidelberg)
Learn more about this promising Nano3Bio achievement from the following short video:
First microalgal chitosan isolated and identified
The research and development team of Greenaltech, a Nano3Bio partner company from Spain, has recently discovered the presence of natural chitosans in certain green microalgae species. With the help of other Nano3Bio consortium partners, Greenaltech is currently working on their characterization. The microalgal chitosans are completely natural sub-stances that do not suffer from any chemical modifications. Moreover, they are of non-animal origin, an important advantage in some industries from a regulatory and marketing perspective. Furthermore, the microalgal chitosan production process is fully controllable as it is performed in closed reactors from the inoculation of the culture media with microalgae to the last chitosan purification step. These advantages, together with the physico-chemical and bioactive properties that are still being elucidated within the Nano3Bio consortium, will be taken into account to select the niche markets in which the microalgal chitosans may have a higher potential of success.
(Nano3Bio consortium partner in charge: Greenaltech)
Get a brief introduction about this process from the following short video:
Chitosan-based hand cream formulation developed
For the cosmetic sector, chitosans are especially interesting due to their antimicrobial and thickening properties. Besides other functions, they can be used as a multifunctional raw material covering the two requirements of microbial stability and viscosity control with just one ingredient. These two functions are determined by variable chitosan characteristics like the molecular size and the side chain distribution. To determine the ideal characteristics necessary for personal care products, the Nano3Bio partner company Cosphatec is testing different chitosan types provided by other consortium partners. By combining different chitosan types, Cosphatec is able to produce a cream formulation in which no other thickener is needed. At the same time, the antimicrobial stability is kept while reducing preservation significantly to a minimum of the usual concentration. Within upcoming research activities of Nano3Bio, it is even aimed to further optimise these results. So far, the heterogeneity of currently available chitosan was one of the main hurdles for establishing frequent usage of this raw material. A benefit of the ability to produce chitosan biotechnologically will be the possible control of the process achievable only through exact knowledge of the underlying biological mechanisms. In this way, Nano3Bio aims to manufacture mass tailored chitosan with properties fine tuned to the desired applications in order to satisfy the corresponding market demand with a reliable and reproducible quality.
(Nano3Bio consortium partner in charge: Cosphatec)
Learn more about Cosphatec's achievement within Nano3Bio from the short video below:
Low-cost protein engineering technology developed
Synthetic biology researchers from academia and industry alike now have access to low-cost genetic material to help identify protein variants. Developed in part through research funding from the European Union’s Nano3Bio project, the GeneArt Strings DNA Libraries tool makes protein engineering, normally an expensive endeavor, attainable to cost-sensitive customers.
The new GeneArt Strings DNA Libraries is complementary to Thermo Fisher’s existing technology currently used for protein engineering by a process known as directed evolution. This method is designed to identify protein variants with improved properties such as enhanced function, better stability or properties that demonstrate novel enzymatic activity. Proteins engineered with enhanced enzymatic activity have many applications in daily life. As an example, enzymes found in many laundry detergents must be engineered to remain stable and active in hot, detergent-rich water, which are very different conditions compared to their natural environment.
The process of screening and engineering proteins is cost- and labor-intensive. The starting point is a DNA library – a collection of variants of the original DNA sequence encoding the protein of interest. These libraries contain thousands to billions of variants and serve to produce the protein variants that enter the screening procedure.
The expensive screening technology used to identify improved protein variants is compounded by the high cost associated with the meticulous process required to produce high-quality DNA libraries. However, research funded through the Nano3Bio project has enabled implementation of novel technologies and a production workflow to lower the cost of library production. Some of the first researchers to receive these new libraries are Nano3Bio consortium members from IQS in Barcelona, who are using them to develop enzyme variants that can produce novel chitosan oligomer types. The Nano3Bio project is funded by the European Union.
Find corresponding information on the GeneArt™ Strings™ DNA Fragments and Libraries website here ...
Caption (as to the below figure): Steps to improve proteins using directed evolution technology. Research by the EU funded project Nano3Bio makes this process, launched by Thermo Fisher Scientific, more accessible.
Protocol for production of chitosans with defined structures
In order to be able to explore the benefits of chitosans in depth, there is the need of obtaining it with a defined structure. In order to do so, one of Nano3Bio’s strategies is to combine enzymatic processes with chemical transformations. For this purpose, the project has dealt with the chemical activation of compounds for the in vitro preparation of controlled chitooligo-saccharide polymers (organic compounds formed by the repetitive linkage of small molecules called monomers) by certain (enzyme-catalyzed) reactions. Materials readily available from natural resources are transformed through chemical reactions into monomers designed to self-condense in a well-defined manner when reacting in the presence of an engineered enzyme. In order to obtain the desired monomers, specifically developed conditions are applied onto the starting materials followed by a treatment that allows the isolation of the product. Thus, other minor impurities that could be detrimental for the quality of the polymer or could even prevent the polymerization reaction are removed from the monomers. These reaction conditions developed during the project do not only allow the preparation of simple molecules, but also of structurally more complex compounds that may confer special properties to the polymers prepared from them. Enantia, a Nano3Bio partner company seated in Barcelona, mainly executed these activities.
(Nano3Bio consortium partner in charge: Enantia)
Learn more about this Nano3Bio achievement in the following short film:
Learn more about chitosans as important renewable biomaterials
Huge Nano3Bio potentials
While the oil is slowly but surely running out, renewable resources are becoming increasingly important. In future, the biological production of raw materials has to play an even greater role to meet the needs in a more efficient manner, with less resource consumption and environmental pollution. Our international consortium of researchers and companies now rises to this challenge. Learn more ...
Manifold benefits and connecting factors
The potential of functional bio-polymers – Nano3Bio's main object of investigation and engineering – is evident. Nano3Bio provides significant chances as well as natural connections to a wide variety of target groups and application fields. Science, industry, society and presumably also the environment will benefit. Learn more ...