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N Takaya

Publications and source records attributed to N Takaya.

29 records · Page 2Linked to original sources
PubMed1997

Purification and characterization of a flavohemoglobin from the denitrifying fungus Fusarium oxysporum.

A flavohemoprotein was purified to homogeneity from the denitrifying fungus Fusarium oxysporum. The purified protein existed as a monomer with a molecular weight of 44 kDa. It was purified in an oxidized form and exhibited the absorption maxima at 401, 540 and 643 nm in its resting form, and at 434 and 555 nm upon reduction with dithionite, respectively. The protein contained 0.5 mol protoheme/mol and 1.1 mol FAD/mol, respectively. When the resting flavohemoprotein was aerobically incubated with NAD(P)H, it was converted to a spectral species that is spectrally very similar to oxyhemoglobins. These properties are characteristics of flavohemoglobins (FHb) of Alcaligenes eutrophus, Escherichia coli, and baker's yeast. Further the amino terminal amino acid sequence of the protein of F. oxysporum was similar to those of these FHbs. These results suggest that the isolated flavohemoprotein of F. oxysporum would be a counterpart of the proteins in the FHb family.

Amino Acid Sequence↗
PubMed1996

Cloning and characterization of the Rhizopus niveus leu1 gene and its use for homologous transformation.

The Rhizopus niveus leul gene was cloned using a 2-kb HindIII fragment of the leuA gene of Mucor circinelloides as a hybridization probe. The coding region of this gene comprises 1890 bp that encode a putative protein of 630 amino acids. The predicted amino acid sequence is similar to those of other alpha-isopropylmalate isomerases (alpha-IPMIs), showing 79.4%, 71.7%, and 60.6% identity with the Mucor circinelloides LeuA, Phycomyces blaskesleeanus Leu1, and Ustilago maydis Leu1, respectively. The vector pRN1, which contains a 3.7-kb BamHI-SacI fragment of the R. niveus leul gene, could complement leucine auxotrophy of R. niveus M-37 (leu1-), suggesting that the gene codes for alpha-IPMI of R. niveus. Both the transformation frequency and mitotic stability with pRN1 were higher than those with leuA gene-containing vectors. It was found by using the leul gene that the autonomously replicating sequence (ARS) of P. blakesleeanus partially functions as an ARS in R. niveus.

Amino Acid Sequence↗
PubMed1995

Analysis of the 3-phosphoglycerate kinase 2 promoter in Rhizopus niveus.

Promoter analysis was performed on the Rhizopus niveus 3-phosphoglycerate kinase 2-encoding gene (pgk2), one of the two pgk genes (pgk1 and pgk2) from this filamentous fungus sequenced so far. Deletion mutants of the promoter region were fused to the Escherichia coli uidA gene (which codes for beta-glucuronidase; GUS), and introduced into R. niveus to measure the intracellular GUS activities of the transformants. Deletion of the sequence between nt -174 to -133 (numbers indicate the position from the putative translation start codon) caused a significant decrease in the ratio of the GUS activity of the transformant cultured in glucose medium compared to that in glycerol medium. In this region, a 21-nt sequence which is well conserved between pgk1 and pgk2 is present. When it was inserted into the promoter region of the uninducible gene encoding RNase Rh of R. niveus, ligated in front of uidA and introduced into R. niveus, the GUS activity of the transformant was greatly induced by glucose, but less by glycerol. We therefore suggest that the 21-nt sequence is a glucose-inducible transcriptional activator of R. niveus. This is the first report on a transcriptional activator in zygomycetes.

Base Sequence↗
PubMed1995

Cloning of the Rhizopus niveus pyr4 gene and its use for the transformation of Rhizopus delemar.

We have cloned a pyr4 gene encoding orotidine-5'-monophosphate decarboxylase of the filamentous fungus Rhizopus niveus. The pyr4 gene of R. nivens has an open reading frame composed of 265 amino-acid residues and has two putative introns. We have also isolated a pyr4 mutant of Rhizopus delemar from 5-fluoroorotic acid-resistant mutants and transformed it with the pyr4 gene of R. niveus as a selectable marker. Introduced DNA was integrated into the chromosome in a multiple tandem array. The mitotic stability of the introduced DNA was increased by a repeated sporulation process. The expression of the Escherichia coli beta-glucuronidase gene in R. delemar was successfully obtained under the control of the pgk2 gene promoter of R. niveus by co-transformation with the pyr4 gene.

Amino Acid Sequence↗
PubMed1994

Cloning and characterization of two 3-phosphoglycerate kinase genes of Rhizopus niveus and heterologous gene expression using their promoters.

Two 3-phosphoglycerate kinase genes (pgk1 and pgk2) were cloned from Rhizopus niveus. It was deduced that both pgk genes have two introns. They have open reading frames of 1,355 bp and 1,356 bp, and code for proteins of 417 and 416 amino acids, respectively. The first introns of both genes are located at similar positions as those of pgk genes from other fungi based on the deduced amino-acid sequences of PGK proteins. The position of their second introns was similar to that of the seventh intron of the human pgk gene. The deduced amino-acid sequences of PGK proteins show high identity (64.8-72.2%) to those of PGKs of other filamentous fungi. When the promoters of each of the pgk genes were fused to the E. coli beta-glucuronidase (GUS) gene and introduced into R. niveus, significant GUS activities were detected in the cell lysates of the transformants, suggesting that GUS protein was expressed under the control of both pgk gene promoters in R. niveus. GUS activity was induced by glucose but not by glycerol, indicating that expression of R. niveus pgk genes was regulated by the carbon source.

Bacterial Proteins↗
PubMed1994

Isolation and characterization of two chitin synthase genes from Aspergillus nidulans.

Two chitin synthase genes, designated chsA and chsB, were isolated from Aspergillus nidulans with the Saccharomyces cerevisiae CHS2 gene as the hybridization probe. Nucleotide sequencing showed that chsA and chsB encoded polypeptides consisting of 1013 and 916 amino acid residues, respectively; the hydropathy profiles of the enzymes were similar to those of other fungal chitin synthases. Northern analysis indicated that both genes were transcribed, suggesting that cellular chitin in A. nidulans is synthesized by at least two chitin synthases. For examination of the roles of the chitin synthase genes in cell growth, gene disruption experiments were done. The chsA disruptant grew as well as the wild-type strain, but the chsB disruptant had severe growth defects that could not be overcome by the addition of 1.2M sorbitol as an osmotic stabilizer. These findings suggested that chsB but not chsA is essential for hyphal growth.

Amino Acid Sequence↗
PubMed1993

[A case of a right bronchial foreign body (rice cake)].

We had a case of a bronchial foreign body (rice cake). A 73-year-old male with severe dyspnea was taken by emergency car. His right breathing sound was very weak, but chest X-ray showed no changes. We diagnosed it as right bronchial foreign body and we could endoscopically remove the foreign body with basket-forcep successfully. We could save his life.

Aged↗
PubMed1992

Purification of two chitinases from Rhizopus oligosporus and isolation and sequencing of the encoding genes.

Two chitinases were purified from Rhizopus oligosporus, a filamentous fungus belonging to the class Zygomycetes, and designated chitinase I and chitinase II. Their N-terminal amino acid sequences were determined, and two synthetic oligonucleotide probes corresponding to these amino acid sequences were synthesized. Southern blot analyses of the total genomic DNA from R. oligosporus with these oligonucleotides as probes indicated that one of the two genes encoding these two chitinases was contained in a 2.9-kb EcoRI fragment and in a 3.6-kb HindIII fragment and that the other one was contained in a 2.9-kb EcoRI fragment and in a 11.5-kb HindIII fragment. Two DNA fragments were isolated from the phage bank of R. oligosporus genomic DNA with the synthetic oligonucleotides as probes. The restriction enzyme analyses of these fragments coincided with the Southern blot analyses described above and the amino acid sequences deduced from their nucleotide sequences contained those identical to the determined N-terminal amino acid sequences of the purified chitinases, indicating that each of these fragments contained a gene encoding chitinase (designated chi 1 and chi 2, encoding chitinase I and II, respectively). The deduced amino acid sequences of these two genes had domain structures similar to that of the published sequence of chitinase of Saccharomyces cerevisiae, except that they had an additional C-terminal domain. Furthermore, there were significant differences between the molecular weights experimentally determined with the two purified enzymes and those deduced from the nucleotide sequences for both genes. Analysis of the N- and C-terminal amino acid sequences of both chitinases and comparison of them with the amino acid sequences deduced from the nucleotide sequences revealed posttranslational processing not only at the N-terminal signal sequences but also at the C-terminal domains. It is concluded that these chitinases are synthesized with pre- and prosequences in addition to the mature enzyme sequences and that the prosequences are located at the C terminal.

Amino Acid Sequence↗
PubMed1991

[6 cases of contralateral atelectasis immediately after thoracotomy in lateral position--its mechanism and treatment].

Six cases of atelectasis developing in the unopened contralateral lung immediately after thoracotomy in the lateral position were presented, and its cause and treatment were discussed. Atelectasis due to retention of secretion showed various atelectatic X-ray shadows which differed in size by the segment or lobe unit. On the other hand, cases of atelectasis whose occurrence was surmised to be related to the lateral position and anesthesia showed mainly infiltrative shadows which extended from the outer region of the middle-lower lung field to the basal segments of the lung, although some cases were accompanied by atelectatic shadows of various size in terms of the segment or lobe unit. Therefore in order to prevent atelectasis, it is desirable to move sputum by aspiration and appropriately apply ventilation, intermittent pressurization and PEEP, while paying careful attention to the unopened lung. In the treatment of atelectasis, selective endobronchial pressurization using a bronchofiberscope equipped with a cuff was effective. It was also very effective to place an endotracheal tube for ordinary anesthesia near the target bronchus with the aid of a bronchofiberscope and perform selective endobronchial pressurization via that tube.

Aged↗