Purification and Properties of Acid Stable Xylanases from Aspergillus kawachii.

Five extracellular endo-xylanases were recognized in the culture broth of shochu koji mold (Aspergillus kawachii, IFO 4308), and three major xylanases (XylA, XylB, and XylC) were purified and characterized. The molecular masses of XylA, XylB, and XylC were 35,000, 26,000, and 29,000, and isoelectric points were pH 6.7, 4.

Purification and characterization of an antibiotic substance produced from Rhizopus oligosporus IFO 8631.

We obtained a purified antibiotic protein from the submerged cultivation broth of Rhizopus oligosporus IFO 8631 by using CM-Cellulofine chromatography and HPLC. The antibiotic did not show a broad spectrum of activity, but it was very active against some of the Bacillus species, especially against Bacillus subtillis (B. natto) at a very low concentration (less than 1 ppm).

Cloning and molecular characterization of the acetamidase-encoding gene (amdS) from Aspergillus oryzae.

We have isolated an acetamidase-encoding gene (amdS) from Aspergillus oryzae by heterologous hybridization using the corresponding Aspergillus nidulans gene as a probe. The gene is located on a 3.5-kb SacI fragment and its nucleotide (nt) sequence was determined.

Nucleotide sequence and expression of the glucoamylase-encoding gene (glaA) from Aspergillus oryzae.

The glucoamylase-encoding gene (glaA) from Aspergillus oryzae was cloned using its cDNA as a probe, which had been isolated previously. From comparison of nucleotide (nt) sequences of genomic clones with its cDNA, the glaA gene was found to contain four short putative introns, 45-56 nt in length. The A.

Characterization of Rarobacter faecitabidus protease I, a yeast-lytic serine protease having mannose-binding activity.

Rarobacter faecitabidus protease I, a yeast-lytic serine protease, was characterized in order to elucidate the mechanism of lysis of yeast cells by this enzyme. The N-terminal amino acid sequence of the enzyme was found to be homologous to those of Lysobacter enzymogenes alpha-lytic protease and Streptomyces griseus proteases A and B around the catalytic His residue, showing that it is a mammalian type serine protease. In a study of its substrate specificity, it preferentially hydrolyzed the ester of alanine among amino acid p-nitrophenylesters.

Cloning and nucleotide sequence of the KHS killer gene of Saccharomyces cerevisiae.

A 5.3-kbp fragment of the KHS gene was cloned from a genomic bank of Saccharomyces cerevisiae No. 115 constructed with an E.

Cloning and nucleotide sequence of the genomic ribonuclease T2 gene (rntB) from Aspergillus oryzae.

Using synthetic oligonucleotide probes, we have cloned a genomic DNA sequence encoding a ribonuclease (RNase) T2 gene (rntB) from Aspergillus oryzae on a 4.8 kb HindIII fragment. DNA sequence analysis of the RNase T2 revealed the following: (1) The gene is arranged as five exons and four introns; (2) The deduced amino acid sequence contains 239 amino acid residues of the mature enzyme.

The glucoamylase cDNA from Aspergillus oryzae: its cloning, nucleotide sequence, and expression in Saccharomyces cerevisiae.

A cDNA for Aspergillus oryzae glucoamylase was cloned, using oligodeoxyribonucleotide probes derived from amino sequences of peptide fragments of the enzyme. The glucoamylase cDNA, when introduced into Saccharomyces cerevisiae, directed the secretion of active glucoamylase into the culture medium. The complete nucleotide sequence of the cDNA contained an open reading frame encoding 612 amino acid residues.

Genetic engineering of a sake yeast producing no urea by successive disruption of arginase gene.

Urea is reported to be a main precursor of ethyl carbamate (ECA), which is suspected to be a carcinogen, in wine and sake. In order to minimize production of urea, arginase-deficient mutants (delta car1/delta car1) were constructed from a diploid sake yeast, Kyokai no. 9, by successive disruption of the two copies of the CAR1 gene.

Chromosomal transformation in Saccharomyces cerevisiae with DNA isolated by pulse field gel electrophoresis.

We isolated and purified yeast chromosome DNA molecules using pulse field gel electrophoresis (PFG). The isolated DNA had nearly the same size as the native chromosomal DNA on PFG. We could directly transform Saccharomyces cerevisiae yeasts with it, and obtain transformants that were selected by complementation of several markers.

Cloning and nucleotide sequence of the KHR killer gene of Saccharomyces cerevisiae.

The KHR gene cloned from a genomic library was on 4.7-kbp DNA fragment and was inserted into YCpG11 vector (KHR-YCp) and YEp vector (KHP-YEp). Transformants with KHR-YEp could secrete 3-4 times as much killer toxin into the media as the donor strain.

Isolation and properties of a chromosome-dependent KHR killer toxin in Saccharomyces cerevisiae.

A strain of the yeast Saccharomyces cerevisiae coding for KHR on the chromosome secreted a toxin that kills sensitive yeasts. The transformants of multicopy vectors carrying the KHR gene could secrete 3-4-fold the killer toxin of the donor strain. This toxic substance was purified 80-fold in specific activity from the culture filtrate by gel filtration and hydrophobic column chromatography.

Mutants of Saccharomyces cerevisiae with defective vacuolar function.

Mutants of the yeast Saccharomyces cerevisiae that have a small vacuolar lysine pool were isolated and characterized. Mutant KL97 (lys1 slp1-1) and strain KL197-1A (slp1-1), a prototrophic derivative of KL97, did not grow well in synthetic medium supplemented with 10 mM lysine. Genetic studies indicated that the slp1-1 mutation (for small lysine pool) is recessive and is due to a single chromosomal mutation.

Dynamic aspects of vacuolar and cytosolic amino acid pools of Saccharomyces cerevisiae.

By using the Cu2+ method (Y. Ohsumi, K. Kitamoto, and Y.

Solid-state ethanol fermentation by means of inert gas circulation.

A new method for solid-state ethanol fermentation (the SSEF system) was experimented on for the ethanol production from solid starchy materials, where a packedbed-type fermentor was used. Both cultivation of Aspergillus saitoi and enrichment of a saccharifying enzyme were effective for hydrolysis of the starch. Ethanol production was set in by a form of parallel fermentation using a respiration-deficient mutant of Saccharomyces cerevisiae.