Abstract: By this invention, for the first time, a method for high-efficiency site-specific genetic engineering, utilizing either native or heterologous CRISPR-Cas9 systems, in the anaerobic bacterium Clostridium pasteurianum, is provided. Application of CRISPR-Cas9 systems has revolutionized genome editing across all domains of life. Here we report implementation of the heterologous Type CRISPR-Cas9 system in Clostridium pasteurianum for markerless genome editing. Since 74% of species harbor CRISPR-Cas loci in Clostridium, we also explored the prospect of co-opting host-encoded CRISPR-Cas machinery for genome editing. Motivation for this work was bolstered from the observation that plasmids expressing heterologous cas9 result in poor transformation of Clostridium. To address this barrier and establish proof-of-concept, we focus on characterization and exploitation of the C. pasteurianum Type CRISPR-Cas system.

Abstract: By this invention, for the first time, a method for high-efficiency genetic transformation of the anaerobic bacterium Clostridium pasteurianum is provided. Clostridium pasteurianum is a bacterium of substantial industrial importance, due to its selectivity and high productivity of the biofuel and biochemical n-butanol, and its ability to grow on a wide variety of inexpensive substrates. Notable among the substrates that it can utilize as a sole source of carbon and energy is glycerine, which is produced in increasing quantities globally as a by-product of biodiesel processing. The industrial exploitation of Clostridium pasteurianum has previously been impeded by the lack of genetic engineering tools for this bacterium. This invention provides such tools for the first time. Included in the invention is a means for protecting newly introduced DNA from degradation by a restriction enzyme within C. pasteurianum. Then, a detailed protocol is given, which enables high-efficiency transformation of C.

Abstract: Compositions and methods for treating or preventing unwanted immune responses are provided. The compositions relate to novel soluble CD83 (sCD83) polypeptides and nucleic acids encoding such polypeptides, improved (sCD83) formulations, and the use of such polypeptides and formulations in the treatment or prevention of allergy, autoimmune disease and transplant rejection. A sCD83 polypeptide is provided, comprising SEQ ID NO:7 or an amino acid sequence having at least 70% identity to SEQ ID NO:7; wherein one or more of amino acid residues 12, 20, 85 and 92 of SEQ ID NO:7 is an amino acid other than cysteine; and optionally, one or more of amino acid residues 1, 2, 3, 4 and 130 are absent.

Abstract: Biologically-active material, particularly hybridoma cells, is immobilized for mass culture in bioreactors to produce biological products, particularly monoclonal antibodies, by ionically-interacting polycationic groups on chitosan with polyanionic groups on a polyanionic water-soluble polymer. Preferably, the chitosan is chitosan acetate and the polyanionic polymer is sodium alginate. In one embodiment, droplets of a suspension of the biologically-active material and water-soluble polymer are gelled by forming a calcium salt of the water-soluble polymer to form temporary capsules, and a sequestering agent for the calcium ions is added to effect ionotropic gelation of the water-soluble polymer and chitosan to form a semi-permeable membrane around each temporary capsule. In another embodiment, porous beads are formed by exposing droplets of a suspension of the biologically active material and water-soluble polymer to an aqueous solution of chitosan and multiple cations.

Abstract: The invention provides a process for the microbial bioconversion of cereal milling by-products into proteinaceous material for human consumption. The by-products are aerobically fermented in a culture of the fungus Neurospora sitophila in the presence of suitable temperature, pH and nutrient conditions, for a time sufficient to grow microbial biomass protein.

Abstract: Fermentation of a substrate by microorganisms capable of utilizing the substrate for growth is improved by conveying a fermentation medium including the substrate and microoganisms through a hollow tube with the assistance of an internal wall-scraper.

Abstract: The presence of small dissolved quantities of carboxypolymethylenes in culture media producing cellulase or penicillin enhances the fermentation parameters.

Abstract: Fermentation parameters are improved by the addition of certain polymers to the fermentation broth. Small quantities of carboxypolymethylene in Aspergillus niger cultures are particularly effective.