Abstract: A novel plasmid pEAP2, which was constructed from Bacillus sp. 170 chromosomal DNA carrying the gene for extracellular production of penicillinase and a vector plasmid pMB9. A novel microorganism, Escherichia coli HB101(pEAP2) carrying the plasmid pEAP2 and being capable of extracellular production of penicillinase. The method for cultivation of the microorganism is characterized by culturing it in the medium containing NaCl (or KCl) for 16-48 hours. According to this invention, useful high-molecular substances can be produced in a high yield.

Abstract: The present invention provides a method of producing oligosaccharides by the reaction between lactose and .beta.-galactosidase from aspergillus oryzae in which an concentration of lactose in an enzyme reaction mixture is between 50 and 90% (w/v) and a reaction temperature is within the range of 55.degree. C. to the temperature at which the .beta.-galactosidase in the reaction mixture is inactivated. This invention therefore makes it possible to obtain the sugar mixture containing high-purity oligosaccharides, small amounts of unreacted lactose and by produced monosaccharides.

Abstract: A method of controlling the regioselectivity of the glycosidic bond between glycosyl donor and glycosyl acceptor in the enzymatic production of an oligosaccharide compound which either consists of or is a fragment or an analog of the cabohydrate part in a glycoconjugate, by reverse hydrolysis or transglycosidation reactions, is described. The synthesis is carried out in that a donor substance which is a mono- or oligosaccharide or a glycoside thereof, is caused to react, in the presence of a glycosidase, with an acceptor substance which is an O-, N-, C- or S-glycoside consisting of a monosaccharide, oligosaccharide or a saccharide analog and at least one aglycon which is O-, N-, C- or S-glycosidically bonded in 1-position, the .alpha. or .beta.-configuration being selected on the glycoside bond between the glycosyl group and the aglycon in the acceptor substance, and the oligosaccharide compound being separated from the reaction mixture.

Abstract: A method for producing oligosaccharides which are represented by the general formula Gal-(Gal)n-Glc (where Gal is a galactose residue, Glc is a glucose residue, and n is an integer from 1 to 4) which is characterized in that lactose or a lactose-containing substance is treated with at least two kinds of .beta.-galactosidases which are produced by different microorganisms. The present invention provides a method of producing oligosaccharides to obtain sweet saccharide mixture which provide sweetness and add oligosaccharides to food and drinks, with a lower increase in calories than that of conventional additives.

Abstract: A method for increasing the rate of chemical reactions involving the conversion of at least one reactant to at least one product involves contacting the reactant with an appropriate monoclonal antibody under conditions permitting the formation of a complex between the antibody and the reactant. The complexed reactant is converted to the product which is then released from the complex. The monoclonal antibody may employ a cofactor and may be directed to a known substrate of an enzyme. Methods for preparing such monoclonal antibodies are also disclosed.

Abstract: A novel gene coding for thermostable .beta.-galactosidase, novel recombinant DNA in which a DNA fragment containing the above gene is inserted, novel Bacillus subtilis in which the above recombinant DNA is introduced, and novel thermostable .beta.-galactosidase obtained by cultivating the above Bacillus subtilis.

Abstract: A highly sensitive, immunoassay method for determining the amount of an analyte in a sample containing a known analyte in an unknown concentration is provided. Sample; a polypeptide-labeled analog of the analyte, an antibody specific for said analyte, a polypeptide partner capable of non-covalently binding with the polypeptide-labeled analyte to form a complex having catalytic activity, and a substrate capable of being converted to a reporter molecule by the catalytic activity of said complex are brought together in a medium. The polypeptide-labeled analyte analog is capable of competitively binding to the antibody and the polypeptide partner, the antibody inhibiting the formation of a catalytically active complex in the absence of analyte, and the concentration of the antibody, polypeptide partner and polypeptide-labeled analyte are such as to cause varying amounts of analyte to be directly related to the conversion of the substrate to the reporter molecule.

Abstract: The invention relates to a process for the cleavage of peptides and proteins at the methionyl bond using cyanogen chloride.

Abstract: Immobilized enzymes are prepared by reacting an enzyme to be immobilized with an organoclay. The immobilized enzymes are enzyme-organoclay complexes in which the binding is substantially pH independent.

Abstract: Disclosed is a method of increasing the production of enzymes involved in carbohydrate metabolism by bacterium of the species F. arborescens or E. coli. The method involves growing the bacterium on an aqueous nutrient medium containing glucose oxime.

Abstract: A high activity immobilized enzyme composite is prepared by covalently bonding a second enzyme layer to a first enzyme layer that is immobilized to a carrier. The carrier is preferably silica gel which has been activated by treatment with a strong base followed by treatment with a strong acid. The first enzyme layer is covalently bonded to the activated silica gel with an aminosilane and a polyfunctional reactant, and the second enzyme layer is covalently bonded to the first layer with a polyfunctional reactant. Third, fourth and more successive enzyme layers may be covalently bonded. The composite has high activity per unit volume, superior stability and good half-life.

Abstract: Effective disinfection and washing of immobilized lactase from Aspergillus oryzae is carried out after a period of hydrolyzing lactose by treatment of the immobilized lactase with aqueous solutions of pH's 2 to 4 and 6 to 8.7, one or both of which contain a disinfectant. The solution having a pH of 6 to 8.7 may be a phosphate buffered solution. The lactase may be immobilized on a high polymer compound by covalent linkage. The immobilized lactase has outstanding resistance to chemicals and wide pH stability so that decrease of lactase activity is minimized.

Abstract: A biologically engineered plasmid coding for the production of .beta.-glucosidase. The plasmid can be incorporated into various microorganisms to enable the microorganism to digest cellobiose, which is produced from cellulose by an endo- or exocellular cellulase. One particular application is the incorporation of this plasmid into a microorganism which produces ethanol. Preferably, this ethanol producing microorganism is also ethanol tolerant.

Abstract: The invention is concerned with the use of sorbitol to stabilize aqueous solutions of yeast lactases. The sorbitol is conveniently used in high concentration so as to act not only as a stabilizing agent but also as an antimicrobial agent. If desired, an antimicrobial agent other than sorbitol can be incorporated. Both aqueous solutions of yeast lactases containing sorbitol and processes for stabilizing aqueous solutions of yeast lactases by incorporation of sorbitol are claimed.

Abstract: Disinfecting of immobilized enzymes is carried out by contacting the immobilized enzymes with a dilute aqueous solution of at least one substituted diethylenetriamine at a concentration and for a period of time which is sufficient to substantially kill the contaminating microorganisms without significant deleterious effects on the immobilized enzymes. The substituted diethylenetriamine is preferably dioctyldiethylenetriamine or a mixture of dioctyldiethylenetriamine and trioctyldiethylenetriamine.

Abstract: This invention is a method for fusing, within a bacterial host, a gene for a cytoplasmic protein to a gene for a non-cytoplasmic protein, so that the hybrid protein produced is transported to, near, or beyond the cell surface for ease of collection and purification, or for immunological use. The invention also encompasses the fused gene created by the disclosed method.

Abstract: A thermal stable beta-galactosidase enzyme is produced from new strains of Bacillus stearothermophilus. The beta-galactosidase is suitable for hydrolysis of lactose at temperatures of at least 55.degree. C. and is especially suitable for use at temperatures of at least 65.degree. C. For hydrolysis of lactose, the beta-galactosidase enzyme may be in the form of substantially purified enzyme that has been immobilized or immobilized whole cells containing beta-galactosidase enzyme.

Abstract: A process for the release of lactase from yeast cells comprising contacting the cells with an alkyl alcohol or a dialkyl ketone, followed by extraction of the lactase in an aqueous solution at a pH in the range 5.5 to 8.0 is disclosed. Also disclosed is a process for the selective reduction of protease activity in lactase preparations comprising heating an aqueous solution of lactase containing protease impurities in the presence of glycerol. Lactase stability during the heating process can optionally be enhanced by addition of manganous ion.

Abstract: A thermostable lactase is obtained by cultivation of a member of the genus Bacillus in a suitable nutrient medium. The resulting lactase-containing cells or the cell-free enzyme are useful for the hydrolysis of lactose in a variety of substrate media.

Abstract: A disulfide compound of a carrier, having S--S exchange reactivity, of the formulaR--S--S--X.sub.1 --X.sub.2 --A [I]wherein R is 2-benzothiazolyl, X.sub.1 is a spacer group directly bound to the --S--S-- and comprises a plurality of carbon atoms in a straight or branched chain, X.sub.2 is an imide or amide bonding group, and A is an insoluble carrier selected from the group consisting of beads gel agarose residue, gamma-aminopropylated nylon beads residue and a residual group of partially reduced polyacrylonitrile porous granules or fibers.

Abstract: A novel lactase having a molecular weight of about 3.times.10.sup.5, an optimum pH value of about 6.0, an optimum temperature of about 60.degree. C. and at least 1 of the ratio of the activity for hydrolyzing lactose to the activity for hydrolyzing a synthetic substrate: o-nitrophenyl-.beta.-D-galactopyranoside (Lact/ONPG ratio), which is produced by cultivating a microorganism of the genus Bacillus being capable of producing the enzyme, particularly Bacillus circulans LOB 377 (ATCC No. 31382) and isolating from the culture broth. This novel lactase is characteristic in the excellent thermostability and the high Lact/ONPG ratio and hence is useful for treating milk and milk products as well as for preventing diarrhea due to lactose intolerance, especially in babies and infants.

Abstract: Method and compositions are provided for replication and expression of exogenous genes in microorganisms. Plasmids or virus DNA are cleaved to provide linear DNA having ligatable termini to which is inserted a gene having complementary termini, to provide a biologically functional replicon with a desired phenotypical property. The replicon is inserted into a microorganism cell by transformation. Isolation of the transformants provides cells for replication and expression of the DNA molecules present in the modified plasmid. The method provides a convenient and efficient way to introduce genetic capability into microorganisms for the production of nucleic acids and proteins, such as medically or commercially useful enzymes, which may have direct usefulness, or may find expression in the production of drugs, such as hormones, antibiotics, or the like, fixation of nitrogen, fermentation, utilization of specific feedstocks, or the like.

Abstract: A process for producing the enzyme .beta.-galactosidase (lactase) involving a novel microorganism (NCIB No 11259) of unknown genus and for using the enzyme to hydrolyse lactose to glucose and galactose.

Abstract: A new .beta.-galactosidase is now provided which is highly effective to digest lactose and stable at acidic pH values and thermally stable. The .beta.-galactosidase is produced by Penicillium multicolor in high yield and is isolated from the culture medium of this microorganism in an inexpensive and easy way.

Abstract: A thermostable lactase is obtained by cultivation of a member of the genus Bacillus in a suitable nutrient medium. The resulting lactose-containing cells or the cell-free enzyme are useful for the hydrolysis of lactose in a variety of substrate media.