Abstract: The present invention relates to nucleic acid sequences and amino acid sequence for Bacillus subtills disulfide bond isomerases, Dsb1 and Dsb2 and methods for increasing the secretion of heterologous and homologous proteins in gram-positive microorganisms.

Abstract: An in vitro method for production of a protein (polypeptide), characterised in that the mRNA encoding the protein is translated when bound to a solid phase. By allowing for a pause at a terminating stop codon and addition of release factor the final proteon may be obtained in almost pure form. By allowing for pauses and restart at internal codons and replacement of the amino acid mixture used before a pausing with an amino acid mixture that differs with resect to labelling, region-labelled polypeptides/proteins can be accomplished. A labelled polypeptide/protein, characterised in that its amino acid sequence contains one or more regions of two or more labelled amino acid residues in sequence. In a subaspect, the polypeptide/protein has at least one amino acid that is occurring twice and this amino acid differs in labelling at at least two positions.

Abstract: The present invention relates to secretion in Gram-positive microorganisms. The present invention provides the nuclei acid and amino acid sequences for the Bacillus subtilis secretion factor SecG. The present invention also provides means for increasing the secretion of heterologous or homologous proteins in gram-positive microorganisms.

Abstract: The invention provides a biosynthetic gene cluster for mitomycin, for example, a mitomycin biosynthetic cluster from organisms such as Streptomyces, for instance, S. lavendulae, as well as methods of using gene(s) within the cluster to alter antibiotic biosynthesis and to prepare a polyketide synthase.

Abstract: A protein with canonical lysyl-tRNA synthetase activity was purified from Methanococcus maripaludis, cloned, and sequenced. The predicted amino acid sequence of the enzyme indicated a novel class I polypeptide structurally unrelated to class II lysyl-tRNA synthetase reported in eubacteria, eukaryotes, and the Crenarchaeote Sulfobus solfataricus. A similar class I polypeptide was isolated from Borrelia burgdorferi, the causative agent of Lyme disease, and an open reading frame encoding a class I-type lysyl-tRNA synthetase was identified in the genome of Treponema pallidum, the causative agent of syphilis. The B. burdorferi gene encoding tRNALysl was cloned and used to make tRNA in vitro. The fundamental difference between pathogen and host in an essential enzyme suggests that class I-type lysyl-tRNA synthetase provides a target for the development of medical and veterinary therapeutics and diagnostics for Borrelia and other microorganism infections.

Abstract: A protein complex containing 245 kDa and 35 kDa components, designated RAFT1 and RAFT2 (for Rapamycin And FKBP12 Target) interacts with FKBP12 in a rapamycin-dependent manner. This interaction has the pharmacological characteristics expected from the observed in vivo effects of rapamycin: it occurs at low nanomolar concentrations of rapamycin and is competed by excess FK506. Sequences (330 amino acids total) of tryptic peptides derived from the affinity purified 245 kDa RAFT1 reveals striking homologies to the predicted products of the yeast TOR genes, which were originally identified by mutations that confer rapamycin resistance in yeast. A RAFT1 cDNA was obtained and found to encode a 289 kDa protein (2550 amino acids) that is 43% and 39% identical to TOR2 and TOR1, respectively.

Abstract: Crabtree negative organisms such as Kluyveromyces, Pichia, Hansenula and Candida, are used to make selected organic products such as lactic acid. The organisms are cultured in a first culture medium that includes glucose, under conditions that promote cellular respiration. The organisms are then cultured under a second set of conditions that promote production of the selected organic product. The organisms preferably contain an exogenous lactate dehydrogenase gene.

Abstract: This invention related to an isolated nucleic acid fragment encoding a tryptophan biosynthetic enzyme. This invention also related to the construction of a Chimeric gene encoding all or a portion of the tryptophan biosynthetic enzyme, in sense or antisense orientation, wherein expression of the Chimeric gene results in the production of altered levels of the tryptophan biosynthetic enzyme in a transformed host cell. The tryptophan biosynthetic enzymes include anthranilate phosphoribosyltransferase (E.C. 2.4.2.18) from corn (Zea mays), soybean (Glycine max) and wheat (Triticum aestivum); indole-3-glycerol phosphate synthase (E.C. 4.1.1.48) from corn (Zea mays), rice (Oryza sativa), soybean (Glycine max), and wheat (Triticum aestivum); and phosphoribosylanthranilate isomerase (E.C. 5.3.1.24) from corn (Zea mays), rice (Oryza sativa), (and wheat (Triticum aestivum).

Abstract: A protein complex containing 245 kDa and 35 kDa components, designated RAFT1 and RAFT2 (for Rapamycin And FKBP12 Target) interacts with FKBP12 in a rapamycin-dependent manner. This interaction has the pharmacological characteristics expected from the observed in vivo effects of rapamycin: it occurs at low nanomolar concentrations of rapamycin and is competed by excess FK506. Sequences (330 amino acids total) of tryptic peptides derived from the affinity purified 245 kDa RAFT1 reveals striking homologies to the predicted products of the yeast TOR genes, which were originally identified by mutations that confer rapamycin resistance in yeast. A RAFT1 cDNA was obtained and found to encode a 289 kDa protein (2550 amino acids) that is 43% and 39% identical to TOR2 and TOR1, respectively.

Abstract: The present invention is directed to a method for stabilizing the flavor of a fermented malt beverage, most particularly a beer, by the addition of one or more inhibitors, blockers, reducing agents or binding agents that inactivate one or more Maillard reaction intermediates that induce staling of the flavor of fermented malt beverages. In preferred such methods, the agents used are reductase enzymes, especially aldehyde reductases, carbonyl reductases, aldose reductases, oxoaldehyde reductases and most particularly oxidoreductases such as isozymes of Old Yellow Enzyme (OYE;EC 1.6.99.1) (e.g., OYE1 and OYE2 and OYE3). The invention is also directed to the fermented malt beverage prepared by such a method, and to the use during the brewing process of reductase enzymes from naturally occurring sources, including those produced by yeasts, to stabilize the flavor of the resulting beer product and to produce a beer having a stable flavor.

Abstract: The present invention relates to novel mammalian amino acid transporter proteins and the genes that encode such proteins. The invention is directed toward the isolation, characterization and pharmacological use of the human amino acid transporter proteins EAAT1, EAAT2, EAAT3 and ASCT1. The invention specifically provides isolated complementary DNA copies of mRNA corresponding to each of these transporter genes. Also provided are recombinant expression constructs capable of expressing each of the amino acid transporter genes of the invention in cultures of transformed prokaryotic and eukaryotic cells, as well as such cultures of transformed cells that synthesize the human amino acid transporter proteins encoded therein. The invention also provides methods for screening in vitro compounds having transport-modulating properties using preparations of transporter proteins from such cultures of cells transformed with recombinant expression constructs.

Abstract: This invention provides isolated nucleic acid encoding sperm-specific protein kinase A anchoring proteins from mammalian species, recombinant expression constructs encoding said mammalian sperm-specific protein kinase A anchoring proteins, cells transformed with said constructs, and homogenous preparations of these proteins prepared using recombinant genetic techniques. The invention also provides analytic tools such as polyclonal antisera and monoclonal antibodies specific for said sperm-specific protein kinase A anchoring proteins of the invention. The invention also provides methods for isolating nucleic acid encoding mammalian sperm-specific protein kinase A anchoring proteins and recombinant genetic methods for producing said mammalian sperm-specific protein kinase A anchoring proteins of the invention.

Abstract: The present invention relates to novel mammalian amino acid transporter proteins and the genes that encode such proteins. The invention is directed toward the isolation, characterization and pharmacological use of a human amino acid transporter protein termed EAAT4 and genes encoding such a transporter. The invention specifically provides isolated complementary DNA copies of mRNA corresponding to this transporter gene. Also provided are recombinant expression constructs capable of expressing this amino acid transporter gene in cultures of transformed prokaryotic and eukaryotic cells, as well as such cultures of transformed cells that synthesize the human amino acid transporter protein encoded therein. The invention also provides methods for screening in vitro compounds having transport-modulating properties using preparations of transporter proteins from such cultures of cells transformed with recombinant expression constructs.

Abstract: Stabilized ubiquitin-lytic peptide fusion polypeptides and a method of making the same by sub-cloning nucleic acid sequences coding for lytic peptides into a plasmid vector comprising a promoter and ubiquitin polypeptide coding sequence, wherein the ubiquitin polypeptide sequence is linked to the 5′ end of the lytic peptide nucleic acid sequence and is translated as a fusion polypeptide.

Abstract: A DNA sequence encoding a human type V adenylyl cyclase is described. The amino acid sequence of the adenylyl cyclase is also described. This adenylyl cyclase is expressed at increased levels in heart and brain tissue, relative to other tissues.

Abstract: The invention provides a human delayed rectifier potassium channel subunit (DRPCS) and polynucleotides which identify and encode DRPCS. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating or preventing disorders associated with expression of DRPCS.

Abstract: Yeast strains transformed with at least one copy of a gene coding for lactic dehydrogenase (LDH) and further modified for the production of lactic acid with high yield and productivities, are described.

Abstract: The present invention relates to a method of mass-producing lactoferrin polypeptides from yeast which is resistant to lactoferrin polypeptides. The present invention also provides Pichia strains, especially Pichia pastoris KCTC 0500BP, that are resistant to lactoferrin polypeptides.

Abstract: In a method for improving the NADH specificity of preferred NADPH-dependent dehydrogenases, the alkalinity of the enzyme is reduced in the coenzyme docking area by an appropriate change of the amino acid sequence achieved by genetic engineering means. The inventive process is especially used for obtaining short-chain dehydrogenases with coenzyme binding areas on the N-terminus from Lactobacillus brevis and Lactobacillus kefir.

Abstract: Recombinant Myxococcus host cell containing recombinant expression vectors containing epothilone polyketide synthase genes can produce epothilones C and D.