A5010527302- Fungal and yeast genetics research
- Enzyme Production and Characterization
- Biofuel production and bioconversion
- Microbial Natural Products and Biosynthesis
- Fungal Biology and Applications
- Fermentation and Sensory Analysis
- Mycotoxins in Agriculture and Food
- Plant Pathogens and Fungal Diseases
- Food Quality and Safety Studies
- Plant biochemistry and biosynthesis
- Microbial Metabolic Engineering and Bioproduction
- Polyamine Metabolism and Applications
- Peptidase Inhibition and Analysis
- Microbial Metabolism and Applications
- Endoplasmic Reticulum Stress and Disease
- Microbial Metabolites in Food Biotechnology
- Amino Acid Enzymes and Metabolism
- Enzyme Catalysis and Immobilization
- Antifungal resistance and susceptibility
- Studies on Chitinases and Chitosanases
- Plant Disease Resistance and Genetics
- Glycosylation and Glycoproteins Research
- Plant-Microbe Interactions and Immunity
- Protein Hydrolysis and Bioactive Peptides
- Cellular transport and secretion
Tohoku University
2015-2024
Institute for Fermentation
2023
University of Shizuoka
2018
Genomics (United Kingdom)
2009-2017
Hokkaido University
2013-2015
Iwate Medical University
2013
Louisiana State University Health Sciences Center New Orleans
2013
Kanagawa Children's Medical Center
2013
Centre Hospitalier Universitaire Sainte-Justine
2013
Hôpital Robert-Debré
2013
The transformation of Aspergillus oryzae has been achieved with a plasmid carrying the nidulans argB gene coding for ornithine carbamoyltransferase (OCTase). frequency was relatively low (0.7 transformants/μg DNA) but transformed phenotype extremely stable many generations without selective pressure. Southern blot analysis revealed that had occurred by integration multiple tandem copies DNA into host genome through non-homologous recombination. There no evidence existence free in...
Exogenous thiamine regulates Aspergillus oryzae thiA , which is involved in synthesis. One of the two introns its 5′‐untranslated region (5′‐UTR) contains motifs (regions A and B) highly conserved among fungal biosynthesis genes. Deletion either relieved repression by inhibited intron splicing, suggesting that regions B are required for efficient splicing. Furthermore, transcript splicing was essential gene expression. These observations suggest a novel expression regulatory mechanism...
Aspergilli known as black- and white-koji molds which are used for awamori, shochu, makgeolli other food beverage fermentations, reported in the literature A. luchuensis, kawachii, or acidus. In order to elucidate taxonomic position of these species, available ex-type cultures were compared based on morphology molecular characters. kawachii acidus showed same banding patterns RAPD, three species had rDNA-ITS, β-tubulin calmodulin sequences differed from those closely related niger...
During a screening of putative diterpene synthase genes found in public databases using the Aspergillus oryzae expression system, it was that single transformant with ACLA_76850 gene from A. clavatus produced sesterterpene alcohol, ophiobolin F, and three minor hydrocarbons. The has two catalytically independent domains (prenyltransferase/terpene cyclase) which are homologous to those synthase, fusicoccadiene synthase. Coevolution both reaction mechanisms these terpene synthases discussed.
Indole-diterpenes represented by paxilline share a common pentacyclic core skeleton derived from indole and geranylgeranyl diphosphate. To shed light on the detailed biosynthetic mechanism of paspaline-type hexacyclic skeleton, we examined reconstitution machinery in Aspergillus oryzae NSAR1. Stepwise introduction six pax genes enabled us to isolate all intermediates synthesize paxilline. In vitro vivo studies key enzymes, prenyltransferase PaxC cyclase PaxB, allowed elucidate actual...
Genome mining is a promising method to discover novel secondary metabolites in the postgenomic era. We applied Aspergillus oryzae heterologous expression system functionally characterize cryptic bifunctional terpene synthase genes found fungal genomes and identified sesterfisherol gene (NfSS) from Neosartorya fischeri. Sesterfisherol contains characteristic 5-6-8-5 tetracyclic ring modified by cytochrome P450 monooxygenase (NfP450) sesterfisheric acid. The cyclization mechanism was proposed...
Successful treatment of aspergillosis caused by Aspergillus fumigatus is threatened an increasing incidence drug resistance. This situation further complicated the finding that strains resistant to azoles, major antifungal drugs for aspergillosis, have been widely disseminated across globe. To elucidate mechanisms underlying azole resistance, we identified a novel transcription factor required normal resistance in fungi including A. fumigatus, oryzae, and nidulans. fungal-specific Zn2-Cys6...
Abstract Penitrem A is one of the most elaborated members fungal indole diterpenes. Two separate penitrem gene clusters were identified using genomic and RNA sequencing data, 13 out 17 transformations in biosynthesis elucidated by heterologous reconstitution relevant genes. These reactions involve 1) a prenylation‐initiated cationic cyclization to install bicyclo[3.2.0]heptane skeleton (PtmE), 2) two‐step P450‐catalyzed oxidative processes forming unique tricyclic (PtmK PtmU), 3) five...
Abstract The biosynthetic machinery of the first fungal ribosomally synthesized and post‐translationally modified peptide (RiPP) ustiloxin B was elucidated through a series gene inactivation heterologous expression studies. results confirmed an essential requirement for novel oxidases possessing DUF3328 motif macrocyclization, highly unique side‐chain modifications by three (UstCF1F2) pyridoxal 5′‐phosphate (PLP)‐dependent enzyme (UstD). These findings provide new insight into RiPP clusters...
We developed a transformation system for Aspergillus oryzae using the nidulans sC gene encoding ATP sulfurylase as selectable marker. The sC− mutants can be readily isolated by positive selection selenate resistance, thereby niaD− mutant strain of A. was bestowed with mutation. Transformation host (niaD−, sC−) plasmid carrying gave random and multi-copy integrants, while that niaD-carrying occurred mainly single-copy homologous integration events (more than 50% frequency), indicating this...
Macroautophagy is a catabolic process by which cytosolic components are sequestered double membrane vesicles called autophagosomes and sorted to the lysosomes/vacuoles be degraded. Saccharomyces cerevisiae has adapted this mechanism for constitutive transport of specific vacuolar hydrolases aminopeptidase I (Ape1) α-mannosidase (Ams1); cytoplasm vacuole targeting (Cvt) pathway. The precursor form Ape1 self-assembles into an aggregate-like structure in cytosol that then recognized Atg19...
Clustering of biosynthetic genes for producing fungal secondary metabolites, which frequently consist less than ten genes, has been recognized with numerous genomes. The heterologous expression whole in the clusters will therefore produce various types natural products when using a suitable host. We introduced gene cluster biosynthesis diterpene aphidicolin into quadruple auxotrophic host, Aspergillus oryzae, by four different vectors (pTAex3, pPTRI, pUSA and pAdeA) harbor starch-inducible...
A unique highly reducing polyketide synthase (HR-PKS) with a reductase domain was identified in betaenone biosynthetic gene cluster. Successful heterologous expression and characterization of the HR-PKS trans-acting enoyl (ER) provide insights into core structure formation decalin scaffold allow reconstitution machinery.
Reconstitution of the biosynthetic machinery for fungal secondary metabolites in Aspergillus oryzae provides an opportunity both stepwise determination pathways and total biosynthesis natural products. However, to maximize utility reconstitution system, a simple rapid strategy introduction heterologous genes into A. is required. In this study, we demonstrated effective method introducing multiple involved by using expression vectors pUARA2 pUSA2, each which contains two cloning sites. The...
Aspergillus fumigatus is the major filamentous fungal pathogen in humans. Infections associated with A. are often treated azole drugs, but resistance to these antifungal agents increasing. Mortality from aspergillosis azole-resistant fungi extremely high. Previous work has identified transcriptional control of drug target-encoding gene cyp51A as an important contributor . Here, we demonstrate that transcription factor AtrR binds a region promoter alleles this conferring clinically...