Abstract: The present invention provides Streptomyces coelicolor strain A3(2)_M22-2C43 obtained by inducing a point mutation in the base sequence of the DagB gene in a wild-type Streptomyces coelicolor strain A3(2) by UV radiation. Since the Streptomyces coelicolor strain A3(2)_M22-2C43 according to the present invention expresses a DagB mutant enzyme expressing little or no DagB beta-agarase or exhibiting little or no beta-agarase activity, there is no need for separate isolation and purification of DagA enzymes from culture fluid, and the culture fluid of the Streptomyces coelicolor strain A3(2)_M22-2C43 or supernatant thereof may be used to produce, from agar or agarose, neoagarose oligosaccharides with a higher content of neoagarotetraose or neoagarohexaose than that of neoagarobiose.

Abstract: The invention is in the field of biotechnology and involves recombinant DNA technology. It provides means and methods for obtaining an insoluble active fusion protein comprising xylose isomerase activity. More in particular, the invention relates to a method for obtaining active insoluble xylose isomerase, comprising the expression in a host organism of a recombinant gene encoding a fusion protein comprising a xylose isomerase in combination with a PPIase, thereby obtaining the active insoluble xylose isomerase. It also provides recombinant fusion proteins comprising xylose isomerase activity as well as their use in converting xylose to xylulose and glucose to fructose.

Abstract: The present invention is related to an enzyme that allows efficient removal of fructan from grain and vegetable raw material. The enzyme according to the invention produces grain and vegetable material having a fructan content significantly lower compared to that of the starting material.

Abstract: The present invention encompasses methods and compositions for detecting pathogenic bacteria. Additionally, the present invention encompasses methods and compositions for catalyzing the dismutation of superoxide radicals.

Abstract: The present invention relates to the use of nucleic acid molecules coding for a bacterial xylose isomerase (XI), preferably coming from Clostridium phytofermentans, for reaction/metabolization, particularly fermentation, of recombinant microorganisms of biomaterial containing xylose, and particularly for the production of bioalcohols, particularly bioethanol, by means of xylose fermenting yeasts. The present invention further relates to cells, particularly eukaryotic cells, which are transformed utilizing a nucleic acid expression construct which codes for a xylose isomerase, wherein the expression of the nucleic acid expression construct imparts to the cells the capability to directly isomerize xylose into xylulose. Said cells are preferably utilized for reaction/metabolization, particularly fermentation, of biomaterial containing xylose, and particularly for the production of bioalcohols, particularly bioethanol.

Abstract: Various aspects and embodiments herein relate to recombinant proteins with at least one protease recognition sequence that can be inactivated by a cognate protease and methods of preparing such proteins. In some embodiments, recombinant phosphoglucose isomerase (Pgi) proteins are provided. In other embodiments, recombinant phosphotransacetylase (Pta) proteins are provided. In yet other embodiments, recombinant transketolase A (TktA) proteins are provided.

Abstract: The present invention relates to methods of optimization of a protein coding sequences for expression in a given host cell. The methods apply genetic algorithms to optimise single codon fitness and/or codon pair fitness sequences coding for a predetermined amino acid sequence. In the algorithm generation of new sequence variants and subsequent selection of fitter variants is reiterated until the variant coding sequences reach a minimum value for single codon fitness and/or codon pair fitness. The invention also relates to a computer comprising a processor and memory, the processor being arranged to read from and write into the memory, the memory comprising data and instructions arranged to provide the processor with the capacity to perform the genetic algorithms for optimisation of single codon fitness and/or codon pair fitness.

Abstract: The present invention relates to a thermophilic arabinose isomerase and a method of manufacturing tagatose using the same, and more precisely, a gene encoding arabinose isomerase originating from the thermophile Thermotoga neapolitana DSM 5068, a recombinant expression vector containing the gene, a method of preparing a food grade thermophilic arabinose isomerase from the recombinant GRAS (Generally Recognized As Safe) strain transformed with the said expression vector, and a method of preparing tagatose from galactose using the said enzyme.

Abstract: A xylose isomerase (XI) enzyme which exhibits increased activity and affinity for xylose is produced by strain TC2-24 of the rumen bacterium, Prevotella ruminicola. The gene encoding this enzyme may be used to produce improved recombinant yeast capable of utilizing xylose. The recombinant yeast are preferably transformed with heterologous polynucleotide sequences coding both the P. ruminicola XI, and the xylulokinase (XKS) of a Prevotella species. Yeast transformed with the polynucleotide sequences coding both of these XI and XKS exhibit significantly increased xylose utilization and cell growth on a culture medium containing xylose as the sole carbon source, in comparison to yeast transformed with XKS and XI from other sources.

Abstract: There is provided in accordance with embodiments of the invention a method of treating or reducing the effects in a subject of a condition selected from fructose intolerance and impaired fructose metabolism, the method comprising administering to a subject in need of such treatment or reduction an efficacious amount of a glucose isomerase, other than in combination with 5-D-fructose dehydrogenase. Other embodiments are also disclosed. There is provided a method for treating or reducing the effects of fructose intolerance and health problems associated with excessive fructose intake by administration of glucose isomerase. Other embodiments are also disclosed.

Abstract: There is provided in accordance with embodiments of the invention a method of treating or reducing the effects in a subject of a condition selected from fructose intolerance and impaired fructose metabolism, the method comprising administering to a subject in need of such treatment or reduction an efficacious amount of a glucose isomerase, other than in combination with 5-D-fructose dehydrogenase. Other embodiments are also disclosed.

Abstract: There is provided in accordance with embodiments of the invention a method of treating or reducing the effects in a subject of a condition selected from fructose intolerance and impaired fructose metabolism, the method comprising administering to a subject in need of such treatment or reduction an efficacious amount of a glucose isomerase, other than in combination with 5-D-fructose dehydrogenase. Other embodiments are also disclosed.

Abstract: A method of decomposing stubble, which is crop residue left on the ground, and a composition for treating the stubble are disclosed. The method comprises treating the stubble with the composition comprising a) polysaccharide-degrading enzyme; b) wetting agent; and, optionally, c) water and allowing the treated stubble to remain on the ground and decompose.

Abstract: A septum is positioned within a disposable vessel and defines a lower chamber and an upper chamber. The septum includes a plurality of apertures that provide fluid communication between the upper chamber and lower chamber. Compressed gas is introduced in the lower chamber to produce fine bubbles rising up throughout the vessel to produce a mixing and gasification needed for the growth of a biological culture and manufacture of a biological product in a nutrient medium. Adding a binding resin to the upper chamber allows harvesting, separation and purification of biological products in the reactor as a single unit operation.

Abstract: An assembly capable of capturing and purifying expressed biological products during or at the end of a bioreaction cycle is disclosed wherein a binding resin is kept separated from the contents of the bioreactor allowing capturing, harvesting and purification of biological products in a bioreactor; the invention additionally provides means of removing undesirable metabolic products as well as provides for efficient loading of chromatography columns.

Abstract: The invention aims to provide a novel alkaline protease having peculiar properties such as high alkali activity, resistance to surfactants and calcium-dependent thermostability and exhibiting excellent performance in highly alkaline detergents, and a gene coding for the amino acid sequence thereof. There is provided an alkaline protease with such properties that an active pH range is from 5 to 13, an optimum pH is approximately 12.6, an optimum temperature is 70° C., no activity drop by heating is observed up to 65° C. at pH 10 and the optimum temperature and the thermostability are not affected by Ca2+ ions. Specifically, there is provided, for example, an alkaline protease having an amino acid sequence constituting a mature enzyme as represented by SEQ ID NO: 3 or an amino acid sequence resulting from deletion, substitution, situs inversus arrangement, addition or insertion of a part of amino acids thereof, or derived from Alkaliphillus transvaalensis.

Abstract: This invention provides a series of recombinant Thermoanaerobacterium saccharolyticum glucose isomerases with improved catalytic activity and thermostability obtained by using recombinant techniques. These recombinant glucose isomerases comprise amino acid variation including phenylalanine (Phe) at position 139, alanine (Ala) at position 182, serine (Ser) at position 187, and glutamine (Gin) at position 299, and carry at least one additional mutated amino acid at position 87, position 217, position 260 or position 276, and possess a higher catalytic activity than that of the wild-type when using D-glucose as substrate. These recombinant glucose isomerases can be used for direct production of high fructose corn syrup containing 55% [wt] or higher concentration of fructose.

Abstract: Methods and compositions for increased production of amino acids from C. glutamicum using sucrose as a carbon source are described. In one aspect, increased production of L-lysine from C. glutamicum is accomplished by using a strain having a mutation in the ptsF gene encoding fructose-PTS enzyme that attenuates or blocks fructose import into the cell when such strain is grown on media containing sucrose as a carbon source and production is increased by providing glucose isomerase in the fermentation media. The glucose isomerase may be exogenously added or expressed in the strain and exported into the media. In certain embodiments the media also contain an invertase. In another aspect increased production of L-lysine is accomplished by making a C. glutamicum strain having the ptsF mutation and a second mutation in a fructose exporter function. The dual mutation retains imported fructose in the cell.

Abstract: This invention provides a series of recombinant Thermoanaerobacterium saccharolyticum glucose isomerases with improved catalytic activity and thermostability obtained by using recombinant techniques. These recombinant glucose isomerases comprise amino acid variation including phenylalanine (Phe) at position 139, alanine (Ala) at position 182, serine (Ser) at position 187, and glutamine (Gin) at position 299, and carry at least one additional mutated amino acid at position 87, position 217, position 260 or position 276, and possess a higher catalytic activity than that of the wild-type when using D-glucose as substrate. These recombinant glucose isomerases can be used for direct production of high fructose corn syrup containing 55% [wt] or higher concentration of fructose.

Abstract: This invention describes a series of recombinant Thermoanaerobacterium saccharolyticum glucose isomerases having improved catalytic activity and thermostability. The recombinant glucose isomerases can be used for direct production of fructose syrup containing 55 wt % or higher concentration of fructose, or used for the production of fructose syrup containing less than 55 wt % fructose.

Abstract: Process for purifying a recombinant protein including one or a few procedural steps only. The process combines the step of lysis of the host cell, with the purification of the protein of interest, allowing for a rapid and much more efficient process of purification. The conditions used during the purification process are those of a high temperature and a low pH, allowing for thermostable and acid-resistant recombinant proteins to be isolated from a suspension. The invention also relates to purifying recombinant proteins which are fusion proteins, wherein one part of the protein may be selected from an enamel matrix protein, such as amelogenin.

Abstract: The present invention relates to a isolated polypeptide characterised in that it comprises an amino acid sequence having at least 80% identity to a sequence selected from the group consisting of SEQ ID NO 2, SEQ ID NO 21 and SEQ ID NO 22, to polynucleotides encoding such a polypeptide and to the use thereof in the production of fructose syrups.

Abstract: This invention provides a series of recombinant Thermoanaerobacterium saccharolyticum glucose isomerases with improved catalytic activity and thermostability obtained by using recombinant techniques. These recombinant glucose isomerases comprise amino acid variation including phenylalanine (Phe) at position 139, alanine (Ala) at position 182, serine (Ser) at position 187, and glutamine (Gln) at position 299, and carry at least one additional mutated amino acid at position 87, position 217, position 260 or position 276, and possess a higher catalytic activity than that of the wild-type when using D-glucose as substrate. These recombinant glucose isomerases can be used for direct production of high fructose corn syrup containing 55.% [wt] or higher concentration of fructose.

Abstract: There is provided in accordance with embodiments of the invention a method of treating or reducing the effects in a subject of a condition selected from fructose intolerance and impaired fructose metabolism, the method comprising administering to a subject in need of such treatment or reduction an efficacious amount of a glucose isomerase, other than in combination with 5-D-fructose dehydrogenase. Other embodiments are also disclosed.

Abstract: The invention generally relates to polyunsaturated fatty acid (PUFA) polyketide synthase (PKS) systems isolated from or derived from non-bacterial organisms, to homologues thereof, to isolated nucleic acid molecules and recombinant nucleic acid molecules encoding biologically active domains of such a PUFA PKS system, to genetically modified organisms comprising PUFA PKS systems, to methods of making and using such systems for the production of bioactive molecules of interest, and to novel methods for identifying new bacterial and non-bacterial microorganisms having such a PUFA PKS system.

Abstract: This invention provides a series of recombinant Thermoanaerobacterium saccharolyticum glucose isomerases with improved catalytic activity and thermostability obtained by using recombinant techniques. These recombinant glucose isomerases comprise amino acid variation including phenylalanine Phe) at position 139, alanine (Ala) at position 182, serine (Ser) at position 187, and glutamine (Gln) at position 299, and carry at least one additional mutated amino acid at position 87, position 217, position 260 or position 276, and possess a higher catalytic activity than that of the wild-type when using D-glucose as substrate. These recombinant glucose isomerases can be used for direct production of high fructose corn syrup containing 55.% [wt] or higher concentration of fructose.

Abstract: The present invention relates to a thermophilic arabinose isomerase and a method of manufacturing tagatose using the same, and more precisely, a gene encoding arabinose isomerase originating from the thermophilic Thermotoga neapolitana DSM 5068, a recombinant expression vector containing the gene, a method of preparing a food grade thermophilic arabinose isomerase from the recombinant GRAS (Generally Recognized As Safe) strain transformed with the said expression vector, and a method of preparing tagatose from galactose using the said enzyme.

Abstract: This invention describes a series of recombinant Thermoanaerobacterium saccharolyticum glucose isomerases having improved catalytic activity and thermostability. The recombinant glucose isomerases can be used for direct production of fructose syrup containing 55 wt % or higher concentration of fructose, or used for the production of fructose syrup containing less than 55 wt % fructose.

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: This invention provides a series of low-copy number plasmids comprising restriction endonuclease recognition sites useful for cloning at least three different genes or operons, each flanked by a terminator sequence, the plasmids containing variants of glucose isomerase promoters for varying levels of protein expression. The materials and methods are useful for genetic engineering in microorganisms, especially where multiple genetic insertions are sought.

Abstract: The present invention relates to novel subtilase variants exhibiting alterations relative to the parent subtilase in one or more properties including: Wash performance, thermal stability, storage stability or catalytic activity. The variants of the invention are suitable for use in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dishwash compositions, including automatic dishwash compositions.

Abstract: The present invention provides novel nucleic acids, novel polypeptide sequences encoded by these nucleic acids and uses thereof.

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 polynucleotide sequences of pgi genes encoding polypeptide sequences having phosphoglucose isomerase activity from coryneform bacteria, to coryneform bacteria containing such polynucleotides and/or polypeptides, a process for the production of L-amino acids using such polynucleotides and/or polypeptides, and methods of screening and amplifying polynucleotides encoding polypeptide sequences which comprise varying degrees of phosphoglucose isomerase activity.

Abstract: The present invention relates, in general, to a method of producing L-amino acids comprising culturing altered bacterial cells having increased amounts of NADPH as compared to unaltered bacterial cells whereby L-amino acids yields from said altered bacterial cells are greater than yields from unaltered bacterial cells. The invention also relates to a gene encoding phosphoglucoisomerase.

Abstract: The present invention provides novel isolated polynucleotides encoding a novel xylose isomerases capable of catalyzing the conversion of glucose to fructose and nucleic acids encoding for such isomerase. The present invention also provides a method of producing the xylose isomerase enzymes employing DNA encoding for the enzymes, plasmids containing the DNA, and bacteria into which the plasmids have been inserted and which produce the enzymes.

Abstract: The present invention relates, in general, to a method of producing L-amino acids comprising culturing altered bacterial cells having increased amounts of NADPH as compared to unaltered bacterial cells whereby L-amino acids yields from said altered bacterial cells are greater than yields from unaltered bacterial cells. The invention also relates to a gene encoding phosphoglucoisomerase.

Abstract: This invention pertains in part to a method for delivering functional DNA or antigens. The desired DNA is introduced into attenuated bacteria able to enter cells. Once the bacteria is in the cell, antimicrobial agents are introduced such that they enter the mammalian cell and lyse the bacteria thereby allowing the delivery of carried functional DNA or antigens. The advantages of this method and its uses are described.

Abstract: The present invention relates to methods and compositions for the treatment of diabetes involving free radicals. In particular, the present invention is directed to the treatment or prophylactic intervention of diabetes. The present invention demonstrates that MnSOD can play a protective role against cytokine killing, and provides strategies for engineering cell lines as islet surrogates for transplantation therapy of diabetes mellitus. Further, the present invention shows that &bgr;-cell destruction and dysfunction in adipogenic diabetes is mediated via fatty acids. Methods and compositions for ameliorating this disorder are provided herein.

Abstract: The invention provides a method and expression system for enhancing secretion of hyperproduced homologous and heterologous exoproteins in gram-positive bacteria such as Bacillus sp. The method and system comprise overproduction of PrsA protein in a gram-positive bacterial host also overproducing at least one exoprotein of interest. Use of the method and system of the invention results in greatly enhanced secretion of the synthesized exoproteins into the growth medium. Once in the growth medium these secreted exoproteins can be recovered and purified in a straightforward manner.

Abstract: The invention relates to a novel water-insoluble glucose isomerase which is formed by a crystalline enzyme converted to solid form by cross-linking. The invention also concerns a process for the preparation of the novel crystalline glucose isomerase by cross-linking with dialdehyde in the presence of a compound containing at least one amino group, and the use of this novel enzyme preparation as an isomerization catalyst.

Abstract: The invention relates to a novel water-insoluble glucose isomerase which is formed by a crystalline enzyme converted to solid form by cross-linking. The invention also concerns a process for the preparation of the novel crystalline glucose isomerase by cross-linking with dialdehyde in the presence of a compound containing at least one amino group, and the use of this novel enzyme preparation as an isomerization catalyst.

Abstract: Secondary-metabolite biosynthesis genes from actinomycetes, method of isolating them, and their use.The invention concerns secondary-metabolite biosynthesis genes from actinomycetes, a method of isolating secondary-metabolite and, in particular, 6-deoxy-sugar bio-synthesis genes, from actinomycetes using the gene probes strD, strE, strL and strM gene probes from Streptomyces griseus DSM40236, and structurally related genes, as gene probes for detecting the genes snot (coding for amphotheronolide B-dTDP-D-mycosaminyl transferase), snoD (coding for dTDP-D-glucose synthase) and snoM (coding for dTDP-4-keto-6-deoxy-D-glucose isomerase), or one or more secondary-metabolite biosynthesis genes from actinomycetes. The invention also concerns the use of secondary-metabolite biosynthesis genes thus isolated.

Abstract: The present invention describes the cloning and expression of a hyperthermostable xylose isomerase gene in an industrial host and the heat facilitated purification of the enzyme. The hyperthermostable enzyme has unique features sought by the fructose syrup industry.

Abstract: A process for producing high fructose corn syrup from glucose containing syrup, which entails isomerizing the glucose containing syrup through an enzymatic reaction to make high fructose corn syrup, wherein the enzymatic reaction occurs in a gas containing solution which comprises at least one gas selected from the group consisting of noble gas.

Abstract: A method of catalyzing in vitro reactions using seeds containing enhanced amounts of enzymes is disclosed. The method involves adding transgenic, non-wild type seeds, preferably in a ground form, to a reaction mixture and allowing the enzymes in the seeds to increase the rate of reaction. By directly adding the seeds to the reaction mixture the method provides a solution to the expensive and problematic process of extracting and purifying the enzyme. Methods of treatment are also provided whereby a subject lacking a sufficient supply of an enzyme is administered the enzyme in the form of seeds containing enhanced amounts of the enzyme.

Abstract: A process for producing high fructose corn syrup from glucose containing syrup, which entails isomerizing the glucose containing syrup through an enzymatic reaction to make high fructose corn syrup, wherein the enzymatic reaction occurs in a gas containing solution which comprises at least one gas selected from the group consisting of noble gas.

Abstract: The invention relates to a novel water-insoluble glucose isomerase which is formed by a crystalline enzyme converted to solid form by cross-linking. The invention also concerns a process for the preparation of the novel crystalline glucose isomerase by cross-linking with dialdehyde in the presence of a compound containing at least one amino group, and the use of this novel enzyme preparation as an isomerization catalyst. Preferrably the compound having an amino group is an ammonium salt, lysine or tryptophan and the dialdehyde is glutaraldehyde.

Abstract: A xylose isomerase gene from Thermus bacteria, such as Thermos aquaticus (ATCC 27634) and a gene having 60% or more of homology to the nucleotide sequence of Thermus aquaticus xylose isomerase gene of FIG. 1-3. A xylose isomerase from Thermus aquaticus characterized in that the xylose isomerase has the optimal pH of about 7, the stable pH range of from about 6 to 8.5, the optimal temperature of about 95.degree. C. and the molecular weight of about 44,000, and is stabilized with manganese or magnesium. A process for preparing a xylose isomerase comprising transforming a microorganism with a plasmid containing the above gene and a promoter, culturing the transformed microorganism and harvesting the produced xylose isomerase. A process for preparing fructose comprising isomerization of glucose to fructose in the presence of the above xylose isomerase.

Abstract: A method for selecting amino acid residues is disclosed which upon replacement will give rise to an enzyme with an altered pH optimum. The method is specific for metalloenzymes which are inactivated at low pH due to the dissociation of the metal ions. The method is based on altering the pK.sub.a of the metal coordinating ligands or altering the K.sub.ass for the metal binding. New glucose isomerases with an altered pH optimum are provided according to this method. These altered properties enable starch degradation to be performed at lower pH values.