Abstract: This invention is concerned with the specific processing of secreted proteins in genetically modified yeast cells. The yeast KEX1 gene was cloned and the KEX1 product was shown to be a serine protease, evidently a carboxypeptidase B-like protease. A probable site of processing of polypeptides by the KEX1 gene product is at the C-terminus of the .alpha. subunit of the killer toxin, where the mature toxin subunit is followed in the precursor by a pair of basic amino acid residues. Processing likely involves an endoprotease cut following these basic residues, and their subsequent C-terminal trimming by a carboxypeptidase. Consistent with the KEX1 product being this carboxypeptidase is the finding that it is also involved in .alpha.-factor pheromone production. In wildtype yeast, KEX1 is not essential for .alpha.-factor production, as the final hormone repeat in the prepro .alpha.-hormone precursor does not need C-terminal processing to form one copy of the active hormone. However, in a mutant strain where .

Abstract: An acyloxyacyl hydrolase from the human promyelocyte cell line HL-60 has been found to specifically hydrolyze fatty acids form their ester linkages to hydroxy groups of 3-hydroxyfatty acids, the latter being bound in turn to LPS glycosaminyl residues. The hydrolyzed fatty acids may include dodecanoic acid, tetradecanoic acid and hexadecanoic acid. This enzyme showed a molecular weight between about 50,000 daltons and about 70,000 daltons.Altered bacterial LPS substantially without fatty acids bound in ester linkage to hydroxy groups of 3-hydroxyfatty acids covalently linked to a glucosaminyl moiety of LPS lipid A are produced. Since the structure of the lipid A moiety is highly conserved, acyloxyacyl hydrolase may act on LPS of many different pathogenic bacteria (for example Salmonella, Escherichia, Hemophilus, and Neisseria).

Abstract: Exoenzymes, such as proteases, xylanases and amylases, are obtained continuously by cultivation of exoenzyme-producing microorganisms in one step in a fermenter which is operated with continuous flow and in which a deficiency state corresponding to maximal enzyme productivity is effected. Optical density of the culture (as a measure of the biomass density) and exoenzyme concentration in culture can be monitored to control the timing and extent of the deficiency state. It is particularly advantageous to impose an oxygen limitation and to maintain the deficiency state continuously by exerting an effect on the oxygen input.

Abstract: New hybrid plasmids which are useful as vectors in recombinant DNA in which Acremonium chrysogenum or Saccharomyces cerevisiae is used as a host, and microorganisms bearing the hybrid plasmids, are disclosed. The disclosure herein is based on the cloning of autonomous replication sequences (ARS) of A. chrysogenum ATCC 11550 and introduction of the ARS into a shuttle vector of E. coli and Saccharomyces cerevisiae and construction of new types of plasmids based on the ARS.

Abstract: Process for the production of a protein correlated with the diphtheric toxin which comprises culturing in a liquid nutrient medium having a concentration of iron ions of from 0.05 .mu.g/ml to 0.5 .mu.g/ml, at a temperature of from 30.degree. C. to 40.degree. C., in a neutral culturing environment and under aerobic conditions, a microorganism belonging to the Corynebacterium diphtheriae genus with two mutant phages encoding the protein correlated with the diphtheric toxin integrated in a nontandem mode in their chromosomes.

Abstract: Antiviral proteins phytolaccin.sub.1 and phytolaccin.sub.2 were isolated from the higher plant Phytolaccin americana using the technique of immunoaffinity chromatography. Phytolaccin proteins were purified to apparent homogeneity in a rapid and efficient chromatographic procedure utilizing immobilized monospecific anti-phytolaccin antibodies. Immunoaffinity-purified phytolaccin.sub.1 and phytolaccin.sub.2 were determined by denaturing gel electrophoresis to be of approximately 25,600 and 28,400 daltons, respectively. The two immunoaffinity-purified proteins were equally potent inhibitors of eukaryotic cell-free protein biosynthesis and exhibited mostly similar high-order UV derivative spectra. Antibodies against phytolaccin.sub.1 and phytolaccin.sub.2 did not cross-react with the heterologous antigen, indicating a structural dissimilarity between the two phytolaccin proteins.

Abstract: C-terminal .alpha.-amidating enzyme preparations, including preparations AE-I, AE-II, AE-IIa and AE-IIb, from the skin of Xenopus laevis, wherein all components can convert a peptide having a glycine residue at its C-terminal to a C-terminal amidated peptide lacking the glycine residue, and have a common N-terminal amino acid sequence represented by Ser-Leu-Ser---, and AE-I and AE-IIa have a molecular weight of about 39,000, AE-IIb has a molecular weight of about 34,000, and AE-II comprises two components having molecular weight of about 39,000 and 34,000; a process for production of the above-mentioned enzyme preparations; and a process for .alpha.-amidation of a peptide by using the above mentioned enzyme preparations.

Abstract: The present invention relates to an improvement in the alpha-amylase method of hydrolyzing a heated slurry of raw starch into starch fragments, the improvement comprising adding a hydrolytically effective amount of the enzyme, glucoamylase, to the reaction slurry.The present invention further relates to a method for the production of high solids dextrim adhesives from raw starch. In carrying out the invention, an aqueous slurry containing raw starch is subjected to hydrolysis at an elevated temperature by the action of two thermally stable enzymes, alpha-amylase and glucoamylase. Once the viscosity of the reaction slurry is reduced to 1000-2000 centipoise as determined by a Brookfield viscometer at 20 rpm, 100.degree. F., and at 45-55% solids (approximately 2.5 hrs.), the enzymes are then inactivated and the reaction is complete. The rheological properties of the resulting slurry can be adjusted as needed.

Abstract: A method for imparting phage resistance to phage sensitive strands of Streptococcus group N is described. The method involves transferring plasmid encoding for production of a mucoid substance (Muc.sup.+) into the phage sensitive strain. Even if the Muc.sup.+ plasmid is removed by curing at elevated temperatures the strains remain resistant to phage. The resulting resistant strains are novel and are used for fermentations, particularly milk fermentations.

Abstract: Recombinant plasmids containing the larvicidal delta-endotoxin gene were constructed by inserting HindIII fragments of the Bacillus thuringiensis var. israelensis 72-75 Md plasmid into the Escherichia coli vector pUC12. Two recombinants producing a 27-kdal toxin (pIP173 and pIP174) were indentified by screening clones in an E. coli in vitro transcription-translation system. Both recombinants comprised pUC12 and common 9.7 kb HindIII fragment of the B. thuringiensis plasmid. The 27,340 Da polypeptide synthesized in vito from pIP174 transformed into E. coli JM101 and from B. subtilis 168 and spoOJ87 containing the 1.2 kb TaqI fragment from pIP173 was lethal to mosquito larvae.

Abstract: This invention relates to a method for producing carnitine comprising contacting, in a reaction medium, carnitinamide with (A) an amidase capable of hydrolyzing carnitinamide to form carnitine or (B) a microorganism containing said amidase, carnitinamide hydrolase and a method for producing same.

Abstract: A purified culture of Ferroxifunis bagdadii capable of extracting metals from mining heaps, liquefying and desulfurizing coal, and decolorizing textile mill wastewaters.

Abstract: S. ghanaensis DSM 2932 is resistant to gentamicin at up to 20 .mu.g/ml. Total digestion of the genomic DNA with BglII, incorporation of the restriction fragments into a suitable plasmid, and selection using gentamicin results in gentamicin-resistant clones which contain a 7 kb fragment with the gentamicin-resistance gene. The plasmid pPH1JI likewise contains a gentamicin-resistance gene located on a 2.3 kb HindIII-BamHI fragment. These genes are suitable as markers, in particular for Streptomycetes vectors.

Abstract: Human insulin precursors containing the peptide chain B(1-29)-A(1-21) of human insulin and derivatives thereof with a bridging chain connecting the carboxyl terminus of the B(1-29)-chain with the amino terminus of the A(1-21)-chain are prepared by culturing a yeast host transformed with a replicable expression vehicle capable of expressing a DNA-sequence encoding the insulin precursor. The bridging chain is preferably relatively short and contains preferably from 2 to 8 amino acid residues. The bridging chain must not contain two adjacent basic amino acid residues (Lys or Arg) and has one Lys or Arg connected to the amino terminus of the A(1-21)-chain. Human insulin is prepared from the insulin precursors by in vitro conversion.

Abstract: The carB gene is a novel carbomycin resistance-conferring gene isolated from Streptomyces thermotolerans and used to construct a number of cloning vectors for use in Streptomyces and related organisms. One such coloning vector, plasmid pOJ159, can be obtained in S. griseofuscus C581 under the accession number NRRL 18090. S. lividans and S. griseofuscus are the preferred hosts when the carB gene is used to select carbomycin-resistant Streptomyces transformants.

Abstract: A description is given of a genertic engineering process for the preparation of mesophilic microorganisms which contain a hydantoinase active at elevated temperature, and of DNA sequences which code for this enzyme.

Abstract: A novel esterase (molecular weight 54,000.+-.2,000) derived from a microorganism belonging to genus Bacilus is extensively utilizable for organic synthetic reaction, especially for asymmetric hydrolysis of ethyl 2,2-dimethyl-3-(2,2-dichlorovinyl)cyclopropane carboxylate (ethyl permethrate), in the same manner as pig liver esterase.The microorganism suitable for production of the above novel esterase is Bacillus sp. DC-1 (FERM BP-1254).

Abstract: The subject invention concerns a novel enzyme named thioredoxin shufflease, means for preparing the same, and procedures for using thioredoxin shufflease to fold proteins containing disulfide crosslinks. Thioredoxin shufflease is a generic term to define enzymes which have the following characteristics: (a) contain a single reactive thiol group; (b) catalyze the exchange of disulfides in a protein undergoing the refolding process; and (c) are not consumed in the oxidation/refolding process. Specifically exemplified is a thioredoxin shufflease produced from an E. coli thioredoxin gene.

Abstract: Disclosed is a simple method for cultivation of bacteria of the genus Campylobacter with good growth of bacteria which comprises carrying out the cultivation in a sealed container of gas barrier type in which are enclosed a medium inoculated with bacteria and a carbondioxide-generating type oxygen-removing composition packed in a gas permeable packaging material, oxygen concentration and carbon dioxide concentration in the container being adjusted to 0.5-15% and 0.5-22%, respectively within 5 hours from the initiation of the cultivation and these concentrations being kept for at most 72 hours.

Abstract: The carA gene is a novel carbomycin resistance-conferring gene isolated from Streptomyces thermotolerans and used to construct a number of cloning vectors for use in Streptomyces and related organisms. One such cloning vector, plasmid pOJ158, can be obtained in S. griseofuscus C581 under the accession number NRRL 18089. S. lividans and S. griseofuscus are the preferred hosts when the carA gene is used to select carbomycin-resistant Streptomyces transformants.