Abstract: In processes for recovery of biologically active polypeptides from fermentation cultures of recombinant host organisms, cell death is frequently a prerequisite for isolation processing of the recombinant product outside the fermentation vessel. Disclosed are improved methods for effecting efficient host cell death inside the fermentation vessel through uniformly contacting host cells in culture with microbicidal concentrations of benzyl alcohol. Illustratively, E. coli, B. subtilis, and P. aeruginosa cultures are advantageously treated with from 0.5 to 10.0% (v/v) of benzyl alcohol in the absence of pH or temperature changes within the fermentor.

Abstract: The present invention relates to a process for preparing a strain of Bacillus, in the chromosome of which a specific gene has been amplified, in which process: (a) at least one plasmid integration vector bearing the said gene is integrated in the Bacillus chromosome so as to create in the chromosome: at least one DNA sequence, known as an amplifiable unit, which contains at least the said gene and its expression elements and, at each end, two sequences which are identical in the direct sense; the amplifiable unit which codes, furthermore, for a selectable gene; (b) the strains of Bacillus obtained are then selected by culture on a selection medium corresponding to the selectable gene, and the strains are withdrawn which have the phenotype corresponding to the presence of an increased number of copies of the said gene relative to the bacterial population before selection.

Abstract: This invention relates to a composition comprising a substantially pure enzyme capable of degrading arabinoxylan and having a molecular weight of about 45,000. More preferably this invention relates to such an enzyme which has a pH optimum between about 6.5 to about 7.0. The enzyme is derived from bacteria capable of hydroloyzing plant cell wall material, preferably from Bacillus most preferably from Bacillus subtilis. This invention further relates to such a composition which is further capable of selectively dissociating feraxan from a maize cell wall preparation. Additionally, compositions of this invention are unable to degrade Rhodymenia (1.fwdarw.3),(1.fwdarw.4)-.beta.-D-xylan and larch arabino-(1.fwdarw.4)-.beta.-D-xylan.

Abstract: The presence or absence of residues of an antibiotic such as penicillin is rapidly determined in a sample such as milk or meat. The sample is added to a test vessel containing a solidified agar medium containing spores of a microorganism which has high sensitivity for the antibiotic being determined. After addition of nutrients and incubation, presence or absence of the antibiotic is indicated by the extent of growth of the microorganism. The nutrients may be in the form of a tablet which may be placed on the surface of the solidified agar medium prior to use. The tablet may be coated with a layer preventing moisture transport from the medium into the tablet during storage, but allowing nutrient transport under test conditions. The extent of microorganism growth may be determined visually or indicated by an indicator present in the solidified agar medium or the nutrient tablet. The test vessel preferably has a cross-section of 3 to 20 mm and a height of 3 to 30 mm.

Abstract: By giving mammals, fowls, fish, etc. feeds containing Bacillus subtilis C-3102 (FERM BP-1096), an excellent body weight gain and feed efficiency can be obtained.

Abstract: A relatively solid, stable biomass reaction product is provided produced from microorganisms having metal uptake properties when contacted by an aqueous solution containing metal cations. The biomass reaction product is produced by treating cells thereof with a caustic solution, whereby the biomass reaction product after drying is characterized in the particulate state of having substantially enhanced uptake of metal cations from aqueous solutions as compared to the metal uptake property of the microorganism before treatment. The biomass reaction product in the particulate state is preferably immobilized in an insoluble binder.

Abstract: A process for preparing optically active indoline-2-carboxylic acid by an optical resolution, which comprises subjecting a racemic ester of (R,S)-indoline-2-carboxylic acid having the general formula [(R,S)-I] to the action of an enzyme or a microorganism having a stereo-selective esterase activity, which is capable of asymmetrically hydrolyzing the racemic ester [(R,S)-I] to give optically active indoline-2-carboxylic acid having the formula [II*] so as to produce the hydrolysis product, i.e. optically active indoline-2-carboxylic acid [II*] and an unreacted optically active ester of indoline-2-carboxylic acid having the general formula [I*], isolating each optically active form, and further, if necessary, hydrolyzing the obtained optically active ester [I*] to give an optical antipode of the acid [II*].According to the process of the present invention, optically active indoline-2-carboxylic acid with a high optical purity can be prepared in a simple process with a good yield.

Abstract: A process for producing useful quantities of a cyclic hydroxide, such as pyrocatechol from a compound having a benzene ring, such as phenol using a Bacillus is described. The process uses tetracycline to inhibit the modification of the cyclic hydroxide by the Bacillus. Pyrocatechol and other related compounds are commercially useful chemicals.

Abstract: A process for the preparation of a pharmaceutically active compound in a stereospecific form of the formula ##STR1## or a pharmaceutically acceptable salt or ester thereof, like an alkali metal salt or an alkaline earth metal salt or a pivaloyl ester, wherein R.sub.1 represents an optionally substituted aryl group such as a phenyl or naphthyl group optionally included in a heterocyclic ring system, which is optionally substituted, or represents a heteroaromatic ring system containing in addition to carbon atoms one or more atoms selected from nitrogen, sulphur and oxygen, this ring system being optionally substituted, which comprises subjecting a compound of the formula ##STR2## wherein R.sub.

Abstract: A recombinant bacteriophage, a method for producing and selecting the recombinant bacteriophage and a method for heterologous cloning of DNA are disclosed. The recombinant bacteriophage is produced by ligating genetic fragments encoding a desired genetic trait with DNA from a bacteriophage, incubating with DNA from a second Bacillus microorganism prototrophic for a growth requirement, incubating with a host Bacillus auxotrophic for the growth requirement. Transformed host Bacillus are selected by growing the mixture on a growth medium which does not contain the growth requirement and determining the presence of the genetic trait. The recombinant bacteriophage containing the desired genetic trait is recovered from the host Bacillus by induction. Heterologous cloning can be accomplished by incubating a host Bacillus with the recombinant bacteriophage.

Abstract: Uridine is produced by cultivating in a culture medium a uridine-producing microorganism, which belongs to the genus Bacillus and which is deficient in uridine nucleoside phosphorylase activity and is resistant to a pyrimidine analogue, and recovering the accumulated uridine. This method has the advantage of substantially avoiding the by-production of uracil and uridylic acid.

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: Cytidine and/or deoxycytidine are produced at high yields by culturing cytidine deaminase activity-defective microbes of the genus Bacillus, which have resistance to pyrimidine analogs and with the ability to produce cytidine and/or deoxycytidine in a medium.

Abstract: A process for preparing pectin which comprises subjecting a plant tissue containing pectic substances to the action of a microorganism which belongs to the genus Bacillus and possesses an activity liberating pectin from a plant tissue but substantially does not possess an activity of decomposing pectin, or a culture broth or processed material thereof to liberate pectin from said plant tissue and recovering the pectin, which allows to obtain readily a pectin of high molecular weight in high yield.

Abstract: Bacillus strains having reduced levels of extracellular protease are produced by replacing the native chromosomal DNA sequence comprising the gene for an extracellular protease, such as subtilisin, with a partially homologous DNA sequence having an inactivating DNA segment inserted therein. The strains are useful as hosts for the expression and secretion of heterologous polypeptides or proteins.

Abstract: Recombinant DNA containing amylase-coding genes is prepared by cleaving DNA from various donor microorganisms and combining portions of the DNA with the plasmid pUB110. Strains of E. coli or B. subtilis containing the recombinant DNA are grown in fermentation media to produce the amylase enzymes.

Abstract: A method is described for increasing the yield of a product from a microorganism containing a regulatory gene, by altering the microorganism. The method involves introducing into the microorganism at least one structural gene for the product by lysogenizing the microorganism with a recombinant bacteriophage containing the structural gene.

Abstract: A replicable plasmidic expression vector, capable of high levels of expression and secretion of polypeptides in a bacillus is disclosed. The vector contains a DNA sequence comprising the promoter and regulatory regions which control expression and secretion of proteases in a bacillus operably liked to a DNA sequence encoding the amino acid sequence of a polypeptide. The expression vector is particularly useful in the production of B. amyloliquefaciens proteases or other heterologous proteins in B. subtilis.

Abstract: A method for treating postharvest stone fruit to prevent or inhibit brown rot of stone fruit with effective amounts of any of the following active ingredients in a carrier is disclosed: Bacillus subtilis B-3; Bacillus subtilis B-3 in combination with 2,6-dichloro-4-nitroaniline; Bacillus subtilis B-3 in combination with water based wax; and, Bacillus subtilis B-3 in combination with paraffin and mineral oil base.

Abstract: A method and a cloning vector are described for the controlled accumulation of cloned heterologous gene products in Bacillus subtilis. The cloning vector is capable of being replicated in B. subtilis and includes the heterologous gene located and oriented such as to be under the control of an operator, promoter, and ribosomal binding site sequence. The gene codes for a protein which is under the control of a transport mechanism by which the protein is secreted by the B. subtilis. The gene product is recovered from the growth medium for the B. subtilis. The cloning vector is also capable of similar use in other bacteria such as E. coli.

Abstract: Method of producing inosine and/or guanosine by culturing an inosine and/or guanosine-producing mutant of the genus Bacillus which requires adenine for growth and is resistant to an antifolate. Thus, inosine and/or guanosine can be produced in much greater yields, compared with known methods.

Abstract: A relatively solid, stable biomass reaction product is provided produced from microorganisms having metal uptake properties when contacted by an aqueous solution containing metal cations. The biomass reaction product is produced by treating cells thereof with a caustic solution, whereby the biomass reaction product after drying is characterized in the particulate state of having substantially enhanced uptake of metal cations from aqueous solutions as compared to the metal uptake property of the microorganism before treatment.

Abstract: Process for the preparation of difficidin and derivative antibacterial compounds of the formula: ##STR1## where R.sub.a and R.sub.b are members independently selected from the group consisting of hydrogen; alkali metal and alkaline earth metal cations; ammonium; and substituted ammonium; and R.sup.1 is hydrogen or hydroxy.

Abstract: A plasmid selected from(a) a plasmid conferring resistance to tetracyline (Tc.sup.r) and neomycin (Neo.sup.r) on a host, and being built up by the in vitro ligation of a Neo.sup.r non-chimeric plasmid and a Tc.sup.r non-chimeric plasmid,(b) a deletion, insertion or deletion/insertion derivative of a group (a) plasmid, or(c) a rearrangement derivative of a group (a) or group (b) plasmid are disclosed.The host may be a Bacillus, particularly Bacillus subtilis.

Abstract: The present invention relates to an expression and secretion vector which comprises the signal and promoter sequence of the Bacillus cereus (herein "B. cereus") gene which codes for penicillinase and to the construction of vectors and the use of the vectors in the expression and secretion of one or more exogenous polypeptides in microorganisms for example, B. subtilis.

Abstract: A method for the production of a pullulanase-like enzyme possessing an .alpha.-amylase activity from a strain of genus Bacillus subtilis, the produced enzyme being capable of acting on starch to enhance the yield of glucose.

Abstract: A process for preparing a fructoside, especially a fructosyl disaccharide, comprises reacting a fructosyl saccharide such as sucrose or raffinose with an alcohol or aldose in the presence of a fructosyl-transferase, especially one derived from B. subtilis NCIB 11811, 11872 or 11873. In particular, aldose is a compound of the formula ##STR1## in which A represents a hydrogen atom or the group CH.sub.2 X, where X represents a hydrogen atom or an alkoxy group, and the fructosyl disaccharide so formed is halogenated to provide a halosucrose or halogalactosucrose sweetener.

Abstract: The gene coding for a thermostable pullulanase enzyme is incorporated into chimeric plasmids which are inserted into and reproduced by E. coli or B. subtilis host microorganisms. When microorganisms containing the chimeric plasmids are grown in fermentation media, they produce the pullulanase enzyme.

Abstract: A novel microorganism is disclosed which has the identifying characteristics of Bacillus subtilis DSM 2704, including high productivity of alpha amylase.

Abstract: A protein effective in stimulating cell growth activity. The protein has a molecular weight of from 5,000 to about 160,000, is composed predominantly of neutral and acidic amino acids, contains a significant amount of glutamic acid and aspartic acid, and is substantially free of nucleoside phosphotransferase. The protein is useful as wound treatment agent and as promoting agent in the synthesis of DNA. A method of producing the protein and compositions containing the same are also disclosed.

Abstract: A mutant of Bacillus subtilis has been isolated which greatly facilitates gene cloning in this nonpathogenic microorganism. B. subtilis is a known protein secretor and can be used efficiently in commercial operations. Unlike the more commonly used clone-propagating organism E. coli., B. subtilis has the advantage of lacking pyrogenic substances in its cell envelope. However, chimeric plasmids for infection of B. subtilis have been difficult to prepare, and if E. Coli is used as an intermediate host to provide plasmid forms suitable for Bacillus transformation, the B. subtilis treats any E. coli-propagated DNA as foreign and preferentially attacks the insert portion of the plasmid. This attach results in loss of cloned genes and limits the use of B. subtilis as a cloning system. The B. subtilis recipient strain of this invention is, on the other hand, stably and efficiently transformed by E. coli-propagated plasmid DNA at high frequency.

Abstract: A method for producing L-phenylalanine by fermentation which comprises aerobically culturing an L-phenylalanine-producing microorganism in an aqueous culture medium and recovering the L-phenylalanine accumulated in the culture medium, the L-phenylalanine-producing microorganism having been constructed by incorporating a recombinant plasmid DNA, into which a DNA fragment controlling resistance to a phenylalanine antagoinst, obtained from a chromosomal DNA of a mutant of the genus Bacillus resistant to the phenylalanine antagonist, has been inserted, into a recipient strain of the genus Bacillus.

Abstract: A strain of Bacillus subtilis designated APPL-1 and a purified extract of APPL-1 were found to effectively control and inhibit rust on bean plants.

Abstract: In a fermentation procedure for the production of nucleosides i.e. inosine and/or guanosine using an adenine-requiring microorganism, the fermentation is carried out by allowing a source of adenine to be present in the medium in an excess amount over the amount of adenine that would be conductive to a maximum yield of inosine and/or guanosine in aerobic culture using ordinary air, and cultivating the microorganism while an oxygen-rich gas is bubbled into the medium. Thus, inosine and/or guanosine are accumulated in high yield in the fermentation broth.

Abstract: A method of enhancing a fungus-lytic activity of .beta.-1,3-D-glucanase which comprises using said glucanase in the presence of one or more of the activators selected from the group consisting of sodium lauroylsarcosinate, polyoxyethylene alkylphenyl ether, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylenealkyl ether, benzalkonium chloride, ammonium chloride, chlorhexidine glucuronate, methylparaben, propylparaben, trypsin, Pronase.RTM. and Alcalase.RTM..A method of enhancing a fungus-lytic activity of .beta.-1,3-D-glucanase which comprises using two .beta.-1,3-D-glucanases of different origins is also provided.

Abstract: A process for preparing uridine diphosphate-N-acetylgalactosamine, which comprises treating a reaction solution obtained by the enzymatic conversion of uridine diphosphate-N-acetylglucosamine to uridine diphosphate-N-acetylgalactosamine, with uridine diphosphate-N-acetylglucosamine pyrophosphorylase to decompose the remaining uridine diphosphate-N-acetylglucosamine in the solution and then separating therefrom the uridine diphosphate-N-acetylgalactosamine for purification. In one aspect of this invention, it relates to a method for measuring the activity of .alpha.-N-acetylgalactosaminyl transferase characterized by the use of said reaction solution as the substrate for the transferase.

Abstract: The aspartase from a mutant Bacillus subtilis, NRRL B-15536, is produced in relatively high cell yield within a comparatively short time. The enzyme converts fumaric acid to L-aspartic acid stoichiometrically with outstanding selectivity and productivity. The enzyme is stabilized by divalent magnesium ions, 2-mercaptoethanol, and ammonium furmarate, and can be conveniently purified with high recovery.

Abstract: A process for economically producing L-tryptophan which involves culturing aerobically in a culture medium a mutant of the genus Bacillus which is resistant to azaserine and a tryptophan analogue and recovering the L-tryptophan produced which accumulates in the culture medium.

Abstract: A method for the determination of the amount of lipase in a sample comprises the steps of:(a) contacting the sample with a reagent composition comprising:(i) a lipase substrate which is a glycerol triester oil having in one of its two .alpha.-ester positions a long chain alkyl group having at least 8 carbon atoms and, in its two remaining ester positions, short chain alkyl groups such that, if the long chain alkyl group is hydrolyzed, the resulting diester is water soluble; and(ii) an esterase enzyme capable of catalyzing the hydrolysis of a water soluble glycerol diester to glycerol; and(b) detecting the rate at which glycerol is formed.The compositions of the present invention include the substrate and enzyme as defined. The element of the invention comprises a support having thereon the described composition. The invention is useful for determining lipase in samples which contain endogenous glycerol, such as blood serum and other body fluids.

Abstract: A mutant Bacillus subtilis, arising from spontaneous adaptation from a parent without detectable aspartase activity, produces recoverable amounts of aspartase. The micro-organism attains maximum growth and enzyme production within about 8 hours and shows excellent glucose tolerance. Aspartase is at least in part produced constitutively, but L-aspartic acid stimulates further aspartase production. The aspartase so produced is readily released from the cell upon cell wall rupture and shows adequate extra-cellular stability for further concentration and purification.

Abstract: An aqueous enzyme preparation stabilized with an ester of the formula RCOOR' where R is an alkyl of from one to three carbons or hydrogen and R' is an alkyl of from one to six carbons, the ester being in an amount of from 0.1 to about 2.5% by weight.

Abstract: A method for testing microbial interaction with growth affecting substances. A pattern of a microbe containing solution is applied on an interaction plate in a programmed concentration. A solution of a growth interacting substance is applied on the interaction plate in a programmed potency in a pattern which contacts the microbe containing solution to cause contact of the microbes with the growth interacting substance. The resultant plate is incubated for a time sufficient to produce visible microbial colonies on the interaction plate. The potency of a growth interacting substance is determined at any point of interest on the incubated interaction plate by correlating the position of the point of interest with the programmed volume of the growth interacting substance deposited at that point.

Abstract: L-Histidine producing microorganisms which have been constructed by introducing a recombinant plasmid DNA inserted on a chromosomal DNA fragment into a recipient strain of the genus Bacillus. These microorganisms are employed to produce L-histidine in higher than normal yields. The recombinant plasmid DNA inserted is obtained from a donor mutant strain of Bacillus subtilis which is resistant to certain L-histidine antagonists. The resistant plasmid confers the properties of the mutant strain upon the recipient Bacillus subtilis to make its high yield of product even higher.

Abstract: Genetically engineered microorganisms are provided which contain recombinant DNA with an amylase coding gene. Improved yields of amylase enzymes are obtained by cultivating these microorganisms.

Abstract: A biologically pure amylase-negative, asporogenous mutant B. subtilis EE101 (ATCC 39,096) is provided. This host, which contains no amylase-coding gene, is particularly useful as a host in a host-vector system for recombinant DNA work directed toward the production of improved strains of amylase-producing microorganisms.

Abstract: A method of producing inosine and/or guanosine comprising cultivating a microorganism in a medium containing a carbohydrate, in which method a carbohydrate is added either continuously or intermittently to the medium when and in the state that the concentration of the carbohydrate in the medium is less than about 1 percent so that the carbohydrate concentration of the medium is maintained below about 1 percent.

Abstract: A biologically pure strain of asporogenous B. subtilis DE101 (ATCC 39,095) is provided. This strain, which shows a low frequency of reversion to spore formers, is suitable for a host providing a high level of biological containment in a host-vector system for use in recombinant DNA methodology.

Abstract: A biologically pure strain of asporogenous B. subtilis DE100 (ATCC 39,094) is provided. This strain, which shows a low frequency of reversion to spore formers, is suitable for a host providing a moderate to high level of biological containment in a host-vector system for use in recombinant DNA methodology.

Abstract: An enzyme which catalyzes the hydrolysis of glycerol esters is disclosed. The enzyme is specific for alkyl esters wherein the alkyl group has from 1 to 4 carbon atoms inclusive. The enzyme is particularly useful in hydrolyzing a diacetyl glycerol ester. The enzyme is from the microorganism Bacillus subtilis ATCC No. 31954.