In the hands of c-LEcta, natural biodiversity is efficiently explored to find enzymes and strains needed for the development of new biotechnological products and for the optimization of current industrial processes. Biodiversity also offers a tremendous potential to identify new active compounds with a broad spectrum of industrially relevant properties. Using a highly efficient, patent-protected screening system, we exploit this potential to provide customized industrial biotech solutions.
c-LEcta possesses a unique and still growing collection of several thousand pre-qualified, dereplicated cultivable microorganisms from different habitats.
These microorganisms are selected, for example, based on their enhanced production of secondary metabolites. The DNA of these microorganisms is available in ready-to-screen expression libraries. Thus, both the microbial strains and the derived recombinant expression libraries can be screened to identify the desired bioactivities.
In many microbial habitats less than 1% of the microorganisms can be cultivated under laboratory conditions. This means that if only cultivable strains or their derived libraries are screened, a vast potential of the microbial world would be left unexploited.
This hurdle is overcome by using metagenomic libraries from uncultured biodiversity. c-LEcta possesses a large number of ready-to-screen metagenomic expression libraries from different habitats, including very special environments such as sheep rumen, chemical industry-contaminated soil or high-temperature decomposition material.
For library building, patent-protected screening vectors have been developed allowing functional based screenings for intracellular and secreted enzymes in different expression hosts.
The performance requirements for enzymes to be applied in industrial processes are in most cases very different from what nature has made them for. This relates to substrate and product concentration, robustness and substrate scope, for instance. Enzymes therefore need to be adapted to industrial conditions by enzyme engineering, and c-LEcta owns one of the most efficient technology platforms to do this. For enzyme optimization or the development of artificial enzyme activities, we use patent-protected enzyme engineering strategies that are inspired by nature. The same principles that Darwin discovered for natural evolution (mutation and selection) have been transferred to a very efficient, proprietary directed evolution process routinely applied in our labs.
The technology platform for enzyme engineering at c-LEcta was developed over the past 15 years and has since proven its efficiency again and again. In the last 10 years we have conducted more than 50 enzyme engineering projects with a success rate of >90%. The key assets of our technology platform include:
> Fast and parallelized methods along the whole process
> A platform of patent-protected and in-house methods for library creation and screening
> Smart search strategies and proprietary bioinformatics-driven library design
We have a proven track record covering many different enzyme classes and target optimization parameters for industrial applications including: substrate scope, stereoselectivity, stability, process performance, inhibition profile and kinetic parameters. In several projects, our outstanding enzyme engineering platform allowed the development of artificial enzymes with completely new, previously undescribed properties. Our experience allows us to discuss any desired project and to provide substantial outlines assessing enzyme choice, assay system, engineering strategy, timelines and chances of success.
c-LEcta uses microorganisms as miniaturized living factories to produce high-value products. We work mainly with three different production platforms, Escherichia coli, Pichia pastoris and Bacillus sp., to produce the enzymes which are currently the most important product group in our portfolio. Besides, in the last years we also have built up a unique genetic engineering toolbox to use our production strains for the direct fermentative production of small-molecule products like carbohydrates, food and cosmetic ingredients or flavors. Our toolbox comprises antibiotic-free markers, serial deletion or insertion strategies for all three host platforms, collections of genetic elements and a proprietary, recombination-based pathway construction technology.
c-LEcta’s strain engineering philosophy
Two aspects make c-LEcta’s strain engineering strategy unique: 1.) strain and enzyme discovery and engineering are carried out in parallel and directly integrated. Our genetic engineering projects benefit from one of the world-best enzyme development platforms, leading to faster and more successful outcomes. 2.) The combination of omics-guided directed strain engineering approaches with random strain optimization. By making use of off-pathway optimization potentials in parallel to rational approaches, we speed up our projects and deliver better performing strains.
c-LEcta’s enzyme engineering platform allows the rapid development of enzymes with undescribed, artificial activities. Through these, new synthetic pathways can be constructed and realized. For the upcoming second wave of fermentative products made by modern biotechnology this is an essential precondition.
The successful commercialization of biotech products in industrial applications is usually driven by superior economics of the production process. For this, both excellent production strains with unrivalled performances and best-in-class production processes are needed. We mainly differentiate two types of bioprocesses being developed and applied at c-LEcta: 1.) fermentation processes, in which a carbon source (usually carbohydrates) is converted by a microorganism into a valuable product. 2.) biotransformation processes, in which an enzyme or an ensemble of enzymes converts one or several substrates into a product (biocatalysis).
Fermentation is the central production technology used at c-LEcta. We have extensive experience in developing, upscaling and implementing microbial fermentation processes based on our three platforms Escherichia coli, Pichia pastoris and Bacillus sp. Examples of our capabilities range from the high cell density, antibiotic- and animal-product-free intracellular and extracellular production of enzymes to the fermentation of low-price sugars to organic acids with highest productivities and titers in very large industrial scale.
The main application we target with our enzyme development work is the use of enzymes for the synthesis of high-value compounds. In some fields, for instance the synthesis of carbohydrates like disaccharides or oligosaccharides, the use of biotechnology has an enormous economic impact and often faces no competition by chemical catalysis. The parallel development of customized enzymes and the design and optimization of single or multi-enzyme synthesis processes is a unique strength of c-LEcta. In special cases enzymatic reactions can be used in intact cells, e.g. when cofactor or cosubstrate regeneration is needed (whole-cell biotransformation).
For an efficient overall process, a state-of-the-art downstream processing is required to meet the economic and quality targets of the product. At c-LEcta we are very experienced in handling the different complexities of DSP development projects ranging from technical-grade crude products to pharma-grade purities transferred into GMP environments. Depending on the product use and the target specifications, downstream processes include separation, tangential flow filtration, bioburden filtration, buffer exchange, chromatography, crystallization, lyophilization or distillation.