Abstract: It is an object of the present invention to provide a novel hydrolase, which is used when dialkyl 2-vinylcyclopropane-1,1-dicarboxylate is hydrolyzed with an enzyme, so as to efficiently obtain (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropanecarboxylic acid that is useful as an intermediate for synthesizing therapeutic agents for hepatitis C. According to the present invention, there is provided a hydrolase protein, which consists of the amino acid sequence shown in any one of SEQ ID NOS. 2 to 5 and which has activity of catalyzing, at higher selectivity than the protein consisting of the amino acid sequence shown in SEQ ID NO. 1, a reaction of producing (1S,2S)-1-ethoxycarbonyl-2-vinylcyclopropanecarboxylic acid from diethyl 2-vinylcyclopropane-1,1 -dicarboxylate.

Abstract: Therapeutic methods for cancer treatments using a combined prokaryotic-eukaryotic delivery and expression system for the delivery of multiple therapeutic factors via a modified tumor-targeted bacteria. A targeted bacteria-vector system elicits an inter-kingdom dual expression (IKDE) of antitumor agents, in the nucleus or cytoplasm of eukaryotic cells, with priming and maintenance of the vector in the bacterium. The therapeutic factors include small interfering RNAs, tumoricidal proteins, DNA molecules, or a combination thereof. The system provides direct killing of tumor cells and alters the tumor microenvironment by expressing anti-angiogenic factors and cytokines in intracellular and/or extracellular environments. Also provided are methods of using natural exosomes comprising cargoes obtained from the bacterially infected cells. The bacteria-vector system is useful for many types of tumor and cancer as well as recombinant vaccines.

Abstract: The invention provides novel strains Agrobacterium tumefaciens KKP 2039p and Paracoccus alcaliphilus KKP 2040p, the plasmid pSinA and its functional derivative, method for producing bacterial strains capable of chemolithotrophic arsenite oxidation and novel bacterial strains produced by this method. The invention also relates to the composition, comprising the novel bacterial strain or the plasmid pSinA and the use of these novel strains, as well as the method of bioaugmentation of an arsenic contaminated environment, particularly the method for the removal of arsenic from waters.

Abstract: The present invention relates to a method for simultaneous fermentation of pentose and hexose. The present invention modifies the metabolic pathways of a target microorganism in order to enable the target microorganism to rapidly metabolize pentose and hexose at the same time. This present invention simplified the fermentation process, decreased the cost, and increased the efficiency of the fermentation process.

Abstract: A method for preparing a xylose-utilizing strain of Saccharomyces cerevisiae and the Saccharomyces cerevisiae are introduced. The preferred recombinant strain contains multiple copies of integrated xylose metabolic genes, and can rapidly ferment xylose to produce ethanol from synthetic medium and lignocellulosic raw materials. The xylose-utilizing strain is applicable for the cellulosic ethanol production industry and brewing industry.

Abstract: Methods for engineering transgenic organisms that synthesize polyhydroxyalkanoates (PHAs) containing 3-hydroxyhexanoate as comonomer have been developed. These processes are based on genetically engineered bacteria such as Escherichia coli or in plant crops as production systems which include PHA biosynthetic genes from PHA producers. In a preferred embodiment of the method, additional genes are introduced in wild type or transgenic polyhydroxybutyrate (PHB) producers, thereby creating new strains that synthesize 3HH monomers which are incorporated into PHAs. The 3HH monomer preferably is derived in microbial systems using butanol or butyrate as feedstocks, which are precursors of 3-hydroxyhexanoyl-CoA. Pathways for in vivo production of butyrol-CoA specifically encompassing butyryl-CoA dehydrogenase activity are provided.

Abstract: The present invention discloses the use of a mutant Leishmania as a suicidal vaccine wherein the mutant Leishmania is responsive to external signals to become porphyric and commit suicidal cytolysis. The mutant can be selected from natural Leishmania species or constructed by genetic engineering.

Abstract: A transposon-based mutagenesis method for altering DNA in Sorangium and other Myxococcales host cells is provided, along with vectors and transposases for use in the method.

Abstract: The present invention provides methods for directing the evolution of microorganisms comprising the use of mutator genes and growth under conditions of selective pressure. The method discloses mutator genes which can be used in the methods of the present invention and provides ATCC deposits which exemplify the evolved microorganisms produced by the methods.

Abstract: The invention relates to a method for increasing the copy number of a chromosomally integrated expression cassette in a microbial strain without leaving antibiotic resistance markers behind in the strain, the necessary genetic constructs, and the strains resulting from the method of the invention.

Abstract: The present invention provides formaldehyde dehydrogenase genes (FLD) from methylotrophic yeasts. The FLD structural genes confer resistance to formaldehyde and are therefore useful as a selectable marker in methylotrophic yeasts. The FLD promoter sequences are strongly and independently induced by either methanol as sole carbon source (with ammonium sulfate as nitrogen source) or methylamine as sole nitrogen source (with glucose as carbon source). Induction under either methanol, methylamine or both provides levels of heterologous gene expression comparable to those obtained with the commonly used alcohol oxidase I gene promoter (PAOX1). The FLD promoter of Pichia pastoris (PFLD1)is an attractive alternative to PAOX1 for expression of foreign genes in P. pastoris, allowing regulation by carbon (methanol) or nitrogen (methylamine) source within the same expression strain.

Abstract: The present invention provides methods for directing the evolution of microorganisms comprising the use of mutator genes and growth under conditions of selective pressure. The method discloses mutator genes which can be used in the methods of the present invention and provides ATCC deposits which exemplify the evolved microorganisms produced by the methods.

Abstract: The present invention relates to a class of microbial coding sequences the transcription or cotranscription of which is specifically induced during microbial infection of a host. These particular coding sequences or defined regions thereof may be used as probes to identify and isolate microbial virulence genes. The products of these virulence genes will provide potential targets for the development of vaccines or antimicrobial agents.

Abstract: The present invention provides methods and compositions for accessing, in a generally unbaised manner, a diverse genetic pool for genes involved in biosynthetic pathways. The invention also provides compounds which can be identified by cloning biosynthetic pathways.

Abstract: New microorganisms with plasmids stable relative to betaine use. These new microorganisms contain (a) a hybrid plasmid with a DNA fragment that contains a genetic sequence that codes for the use of betaine and (b) a mutation in the chromosomal gene coding for the betaine use.

Abstract: The present invention discloses a selection marker free system which can be used to introduce genetic modifications in bacteria, yeasts and fungi. The system can be employed to introduce or delete desired genes or DNA fragments in the genome of the indicated host species without leaving any undesired DNA i.e. the selection marker used for selection of transformants or other DNA used for cloning. In this way strains have been developed containing only desired genes introduced at desired chromosomal sites. Similarly, desired DNA fragments have been deleted or replaced at desired sites.

Abstract: A positive selection vector is provided for the transformation and screening of Gram positive bacteria and particularly Bacillus sp. for the presence of foreign DNA. The vector comprises a mutant gene encoding a signal peptide processing mutation. Expression of the mutant gene in a Bacillus host cell which lacks the ability to metabolize sucrose is lethal when cells are grown in the presence of sucrose. Foreign DNA may be inserted into the vector so as to inactivate the mutant gene thereby permitting the cells to grow in the presence of sucrose and allowing for facile selection of transformants.

Abstract: A system is described which utilizes a novel system of repressor titration for maintenance of a plasmid useful in gene therapy and production of a recombinant protein. The system utilizes a transformed host cell containing a plasmid including an operator susceptible to binding by a repressor expressed in trans, a first chromosomal gene encoding the repressor, and a second chromosomal gene that is functionally associated with an operator and essential for cell growth, wherein the plasmid is present in the cell in sufficient numbers to titrate the repressor such that the essential gene is expressed, thereby permitting cell growth.

Abstract: An overall process for the preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA) via enzymatic ring expansion activity on penicillin G, using a Penicillium chrysogenum transformant strain expressing modified expandase enzyme.

Abstract: The present invention concerns defective recombinant adenoviruses containing an inserted gene encoding apolipoproteins, pharmaceutical compositions comprising the adenovirus, and their use for the treatment or prevention of pathologies linked to dyslipoproteinemias.