ACIB GmbH | Date: 2017-04-19
The present invention relates to a method for producing chiral -nitro alcohol compounds. The invention relates in particular to an (R)-selective cupin- nitroaldolase, which enantioselectively can catalyze the Henry reaction, wherein an aldehyde or ketone compound is converted to the corresponding -nitro alcohol compound in the presence of a nitroalkane compound and a cupin-nitroaldolase.
ACIB GmbH | Date: 2017-08-02
A method of producing a mutant host cell with altered expression of a gene of interest (GOI) by a temperature-inducible CRISPR/Cas system, which comprises:a) at a first working temperature, introducing into a host cell a CRISPR/Cas system employingi) an inactive pre-guide RNA (pre-gRNA) which is a ribozyme-extended guide RNA comprising a guide RNA (gRNA) that recognizes a predetermined target site at the GOI, terminally extended by at least one self-processing Hammerhead (HH) ribozyme characterized by the presence of a thermosensitive hairpin and a cleavage site adjacent to the gRNA; andii) a RNA-guided Cas endonuclease which recognizes the target site upon hybridizing with the gRNA;b) increasing the first working temperature to a second working temperature, thereby changing the structure of the thermosensitive hairpin and activating the pre-gRNA by self-catalyzed cleavage at the cleavage site to obtain the gRNA, which induces binding of the endonuclease to the target site, thereby altering expression of the GOI.
Agency: European Commission | Branch: H2020 | Program: IA | Phase: BIOTEC-3-2014 | Award Amount: 11.39M | Year: 2015
Oxygen functionalities are key functional groups in many of todays chemicals and materials. The efficient introduction of oxygen-functionalities into raw materials are key chemical transformations in bulk and fine chemicals. Innovative bio-catalytic oxidation routes using molecular oxygen (from air) under benign and mild (pH) conditions such as ambient temperature and pressure can greatly improve the sustainability and economics of processes, but were so far mainly been applied in the pharma segments. In this segment, the enzyme-catalyzed step often represents the highest added value and the high price of the end-product (> 1000/kg) justifies less than optimal enzyme production and limitations in its catalytic efficiency. In order to achieve the widening of industrial application of enzymatic bio-oxidation processes to also larger volume but lower price chemical markets, ROBOX will demonstrate the techno-economic viability of bio-transformations of four types of robust oxidative enzymes: P450 monooxygenases (P450s), Baeyer-Villiger MonoOxygenase (BVMOs), Alcohol DeHydrogenase (ADH) and Alcohol OXidase (AOX) for which target reactions have already been validated on lab-scale in pharma, nutrition, fine & specialty chemicals and materials applications. ROBOX will demonstrate 11 target reactions on large scale for these markets in order to prepare them for scale up to commercial-scale plants. ROBOX is industry-driven with 2 major industrial players and 6 SMEs. It will assess the potential of technologies applied to become platform technologies technologies (multi-parameter screening systems, computational methodologies, plug bug expression systems) for broad replication throughout the chemical industry. The markets addressed within ROBOX represent a joint volume of over 6.000 ktons/year. The introduction of bio-oxidation processes is expected to bring substantial reductions in cost (up to -50%), energy use (-60%), chemicals (-16%) and GHG-emissions (-50%).
Agency: European Commission | Branch: H2020 | Program: MSCA-ITN-ETN | Phase: MSCA-ITN-2015-ETN | Award Amount: 3.91M | Year: 2015
We need to increase the crop yield while reducing pesticide and use of inorganic fertiliser to meet the challenges of world population growth and climate change. Plant endophytic microorganisms can improve plant yield and enhance plant tolerance to abiotic stress as well as to pathogens under experimental conditions, but these effects are often not sufficiently stable for practical application. How do we boost the stability and reliability of the positive effects of endophytes on plants? We need to understand the genetic basis of beneficial interactions between crops and endophytes and extent this basis exhibits phenotypic plasticity at all interaction levels from the cellular to the field environment. This requires increasing our knowledge of the molecular mechanisms underlying the effects of endophytes, including intra and inter-kingdom exchange and distribution of resources (nutrients), signalling and possibly regulation between and inside the partners, the mutual induced production of secondary metabolites and the environmental cues which influence crop-endophyte interactions. The genetic variation and its plasticity in host and microbe will be exploited in to establish crop breeding and inoculum production processes for boosting the establishment and stability of plant-microbe mutualisms to benefit crop development, stress tolerance, pathogen resistance and quality. In this project we will provide fundamental biological as well as practical knowledge about interactions between endophytes and plants. This improved understanding will pave the way for increased use of endophytes to improve sustainability and plant productivity in a reliable way. The participants in this project comprise many of the key institutions and industries working with these problems and provide a uniquely strong consortium to address the key issues. Furthermore, the consortium will train a new generation of scientists who have the insight and skills to continue this task in their careers.
Agency: European Commission | Branch: FP7 | Program: CP-TP | Phase: KBBE.2013.3.3-04 | Award Amount: 6.74M | Year: 2013
Glycosyl transferases (GT) are extremely efficient biocatalysts that can be used for the synthesis of special carbohydrates and glycoconjugates. Famous examples of such products include human milk oligosaccharides that serve as prebiotics, and glycosides of flavonoids with improved stability and solubility. Unfortunately, the large-scale application of GT has been hampered by their low operational stability and by the high cost of their glycosyl donor. This project aims to solve both problems by a combination of enzyme and process engineering. On the one hand, the production of stable biocatalysts will be facilitated by the development of a suitable expression system, the design of optimized variants and the development of immobilized formulations. On the other hand, the production and recycling of nucleotide-activated sugars will be accomplished by exploiting the reaction of sucrose synthase (SuSy). With the help of SuSy, UDP-glucose can be produced from sucrose as cheap and abundant substrate. Furthermore, other nucleotide sugars can be obtained when sucrose analogues are employed. These alternative substrates will be produced here with fructansucrase (FS), an enzyme that can couple fructose to various monosaccharides. However, the activity of both FS and SuSy towards sucrose analogues will have to be improved to become economically viable. In that way, the proposed concept can be developed into a generic procedure, in which the nucleotide moiety can be recycled to establish a constant supply of glycosyl donor. The three enzymatic steps will be optimized and integrated into an efficient process for the production of glycosylated compounds. The economical potential of this technology will be demonstrated by the scale-up of selected reactions at the industrial facilities available in our consortium. The participants in this project comprise 3 universities, 1 research institute and 4 SME with complementary skills and expertise.
ACIB GmbH and University of Vienna | Date: 2014-06-10
The invention relates to an isolated waterproof polymeric nanomembrane comprising pores of different geometric shapes and of a controlled size between 10 and 1000 nm, which is larger than the thickness of the membrane, and a method of producing the same comprising the process steps a. Providing a sacrifice layer on a surface of a solid support; b. Providing a polymerized layer of less than 1000 nm thickness on the surface of the sacrifice layer, by depositing a mixture of a polymer or a polymer precursor with a geometrically undefined pore template which is larger than the thickness of the polymerized layer, optionally followed by polymerization and/or crosslinking; c. Removing the pore template to obtain the polymerized layer with a controlled pore size; and d. Removing the sacrifice layer, thereby separating the solid support from the polymerized layer.
Agency: European Commission | Branch: H2020 | Program: MSCA-ITN-ETN | Phase: MSCA-ITN-2014-ETN | Award Amount: 4.04M | Year: 2015
Chinese hamster (CHO) ovary cells are the production host for a \50 billion /yr biopharmaceuticals market. Current CHO production platforms dates to 1980 and are based primarily on media and process optimisation with little consideration to the optimization of the cellular machinery. Fortunately, with the recent sequencing of the CHO genome, an opportunity has opened to significantly advance the CHO platform. The benefit will be advanced production flexibility and a lower production cost. This ITN graduate training programme - eCHO Systems - will blend conventional molecular, cellular, and synthetic biology with genome scale systems biology training in omics data acquisition, biological network modeling, and genome engineering in three interdisciplinary topics: 1) Acquisition of large scale omics data sets and their incorporation into genome-scale mathematical models 2) Development of genome engineering tools, enabling synthetic biology 3) Application of systems and synthetic biology and genome engineering to improve performance of CHO producers The training projects are supported by 15 industrial participants, which will participate in the research and test the results. ESR training will include intense courses focused on computational systems biology, cell biology, business and entrepreneurship. The three universities bring unique complementary skills in systems and synthetic biology, omics technologies, cytometry, and molecular cell biology which will provide depth and breadth to this training. The eCHO Systems will produce four major outputs: General knowledge to improve the productivity, quality, and efficiency of CHO platform cell lines, new systems models for CHO cells, new CHO cell line chassises generated through synthetic biology approaches, high quality education at the graduate level, and a cadre of interdisciplinary graduates poised to transform biopharmaceutical biotechnology.
ACIB GmbH | Date: 2015-04-22
The invention refers to a library of bidirectional expression cassettes or expression vectors comprising a repertoire of bidirectional promoter sequences, each expression cassette comprising a promoter sequence operably linked to a first gene in one direction, and operably linked to an oppositely oriented second gene in the other direction which is different from the first gene, and bidirectional Pichia pastoris or CHO cells promoter sequences. The invention further refers to a method of screening or selecting a bidirectional promoter suitable for expressing at least two GOI in a host cell and a kit comprisinga) an expression cassette consisting of the first and second genes and a stuffer sequence separating them, which stuffer sequence comprises a recognition site for a type IIS restriction enzyme at both ends;b) the type IIS restriction enzyme;c) and a repertoire of promoter, preferably a promoter library including bidirectional promoters.
ACIB GmbH | Date: 2015-12-16
The present invention relates to an improved recombinant cupin-hydroxynitrile lyase (HNL), which is capable to catalyze the asymmetric cyanohydrin reaction, wherein the recombinant cupin-HNL comprises at least one amino acid substitution, preferably at least three substitutions.
ACIB GmbH | Date: 2015-05-27
The present invention relates to an enhanced method for producing a modified target polypeptide containing at least one non-canonical amino acid. In particular, the present invention relates to an improved fermentation method for producing a modified target polypeptide containing at least one non-canonical amino acid, wherein the host cell is cultivated in a minimal medium comprising a complex amino acid source with a limiting amount of at least one canonical amino acid.