Publications

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diversity, application, and synthetic biology of industrially important aspergillus fungi.the filamentous fungal genus aspergillus consists of over 340 officially recognized species. a handful of these aspergillus fungi are predominantly used for food fermentation and large-scale production of enzymes, organic acids, and bioactive compounds. these industrially important aspergilli primarily belong to the two major aspergillus sections, nigri and flavi. aspergillus oryzae (section flavi) is the most commonly used mold for the fermentation of soybeans, rice, grains, and potatoes. asper ...201728732553
structural features of aspergillus niger β-galactosidase define its activity against glycoside linkages.β-galactosidases are biotechnologically interesting enzymes that catalyze the hydrolysis or transgalactosylation of β-galactosides. among them, the aspergillus niger β-galactosidase (anβgal) belongs to the glycoside hydrolase family 35 (gh35) and is widely used in the industry due to its high hydrolytic activity which is used to degrade lactose. we present here its three-dimensional structure in complex with different oligosaccharides, to illustrate the structural determinants of the broad speci ...201728391618
development of a versatile and conventional technique for gene disruption in filamentous fungi based on crispr-cas9 technology.filamentous fungi represent an invaluable source of pharmaceutically active compounds. the development of versatile methods to genetically manipulate filamentous fungi is of great value for improving the low yields of bioactive metabolites and expanding chemical diversity. the crispr-cas9-based system has become a common platform for genome editing in a variety of organisms. however, recent application of this technology in filamentous fungi is limited to model strains, a versatile method for ef ...201728835711
development of a mouse model of eosinophilic chronic rhinosinusitis with nasal polyp by nasal instillation of an aspergillus protease and ovalbumin.one subtype of chronic rhinosinusitis with nasal polyps (crswnp) is characterized by the development of a t-helper type 2 (th2) response and eosinophilic infiltration. here, we aimed to establish an eosinophilic crswnp murine model, which would be essential to understand the underlying pathogenesis and establish a treatment strategy. c57bl/6 mice were challenged intranasally with a mixture of an aspergillus oryzae-derived protease (ap) and ovalbumin (ova) for 6, 8, or 12 consecutive weeks (12 mi ...201728828551
immobilization of β-galactosidase on surface modified cobalt/multiwalled carbon nanotube nanocomposite improves enzyme stability and resistance to inhibitor.the present study aimed to work out a high yield procedure for immobilization of aspergillus oryzae β-galactosidase on polyaniline cobalt multiwalled carbon nanotubes nanocomposite (pani/co/mwcntnc) by physical adsorption and covalent attachment via glutaraldehyde. the binding was confirmed by scanning and transmission electron microscopy along with fourier transform-infrared spectroscopy. the immobilization yields obtained for adsorbed and cross-linked enzymes were 93% and 97%, respectively. th ...201728735002
highly efficient gene targeting in aspergillus oryzae industrial strains under ligd mutation introduced by genome editing: strain-specific differences in the effects of deleting ecdr, the negative regulator of sclerotia formation.numerous strains of aspergillus oryzae are industrially used for japanese traditional fermentation and for the production of enzymes and heterologous proteins. in a. oryzae, deletion of the ku70 or ligd genes involved in non-homologous end joining (nhej) has allowed high gene targeting efficiency. however, this strategy has been mainly applied under the genetic background of the a. oryzae wild strain rib40, and it would be laborious to delete the nhej genes in many a. oryzae industrial strains, ...201728484116
enzymatic characterization of a novel xaa-pro aminopeptidase xpma from aspergillus oryzae expressed in escherichia coli.xaa-pro aminopeptidases are peptidases responsible for the cleavage of any amino acid n-terminally adjacent to a proline residue. we identified a gene encoding a putative xaa-pro aminopeptidase in the genome of the filamentous fungus aspergillus oryzae (genome database number: ao090701000720) and named this gene xpma. we produced its enzyme in a c-terminally his6-tag-fused form in an escherichia coli expression system and purified it. the purified recombinant xpma (rxpma) showed hydrolysis activ ...201728698148
direct alkylation of amines with primary and secondary alcohols through biocatalytic hydrogen borrowing.the reductive aminase from aspergillus oryzae (aspredam) was combined with a single alcohol dehydrogenase (either metagenomic adh-150, an adh from sphingobium yanoikuyae (syadh), or a variant of the adh from thermoanaerobacter ethanolicus (tesadh w110a)) in a redox-neutral cascade for the biocatalytic alkylation of amines using primary and secondary alcohols. aliphatic and aromatic secondary amines were obtained in up to 99 % conversion, as well as chiral amines directly from the racemic alcohol ...201728671344
free and immobilized aspergillus oryzae sbs50 producing protease-resistant and thermostable phytase.optimization for enhanced phytase production by aspergillus oryzae sbs50 in submerged fermentation was investigated using taguchi design. in first step design, starch, beef extract, magnesium sulphate, ferrous sulphate and tween 80 were identified as significant factors affecting phytase production. these significant factors were further optimized at four different levels using a second taguchi design and were observed that 1% starch, 2% beef extact, 3% tween 80, 0.1% magnesium sulphate and 0.22 ...201728669072
comparative analysis of polyspecificity of the endogenous trna synthetase of different expression host towards photocrosslinking amino acids using an in silico approach.photo-induced covalent crosslinking has emerged as the powerful strategy for analyzing and characterizing the protein-protein interaction and mapping protein 3d conformations. in the last decades, a number of photocrosslinking amino acids have been reported but only a few have been efficiently utilized for photocrosslinking purposes. recently, incorporation of diazirine containing photoactivatable analogs such as photo-methionine, photo-leucine, photo-isoleucine and photo-lysine into target prot ...201728641210
purification and characterization of a newly serine protease inhibitor from rhamnus frangula with potential for use as therapeutic drug.protease inhibitors from plants are well known to be potent inhibitors of the growth of bacteria, fungi, and even certain viruses which make them excellent candidates for use as the lead compounds for the development of novel antimicrobial agents for applications in medicine. in this study, rhamnus frangula was selected as a protease inhibitor source. the maximum recovery of the protease inhibitor against trypsin was recorded in the crude extract made in 0.1 m phosphate buffer (ph 7.0) and isola ...201728597160
comparison between the univariate and multivariate analysis on the partial characterization of the endoglucanase produced in the solid state fermentation by aspergillus oryzae atcc 10124.endoglucanase production by aspergillus oryzae atcc 10124 cultivated in rice husks or peanut shells was optimized by experimental design as a function of humidity, time and temperature. the optimum temperature for the endoglucanase activity was estimated by a univariate analysis (one factor at the time) as 50 °c (rice husks) and 60 °c (peanut shells), however, by a multivariate analysis (synergism of factors) it was determined a different temperature (56 °c) for endoglucanase from peanut shells. ...201728857660
engineered humicola insolens cutinase for efficient cellulose acetate deacetylation.cutinases comprise a family of esterases with broad hydrolytic activity for chain and pendant ester groups. this work aimed to identify and improve an efficient cutinase for cellulose acetate (ca) deacetylation. the development of a mild method for ca fiber surface deacetylation will result in improved surface hydrophilicity and reactivity while, when combined with cellulases, a route to the full recycling of ca to acetate and glucose. in this study, the comparative ca deacetylation activity of ...201728488758
consumption of an acid protease derived from aspergillus oryzae causes bifidogenic effect in rats.a marked elevation in the abundance of bifidobacterium was found in the cecum of rats that were fed a high-fat diet supplemented with an amano protease preparation (derived from aspergillus oryzae). the protease preparation contains several digestive enzymes, including acid protease (acp), alkaline protease, and amylase. we hypothesized that the elevation in the abundance of bifidobacterium by amano protease preparation is associated with the digestive enzymes involved in the protease preparatio ...201728821318
fungal endocarditis due to aspergillus oryzae: the first case reported in the literature.infective endocarditis (ie) is a severe disease with high mortality and morbidity. prosthetic valve endocarditis is a life-threatening complication which can occur in less than 10% of patients with valve prosthesis. a fungal etiology of ie is rare and accounts for only 2-4% of all case of endocarditis, but is associated with a higher mortality and morbidity. herein is reported the first case of fungal endocarditis of aortic valve prosthesis due to aspergillus oryzae in a 67-year-old caucasian ma ...201728820551
biochemical characterization of aspergillus oryzae recombinant α-l-rhamnosidase expressed in pichia pastoris.an α-l-rhamnosidase-encoding gene from aspergillus oryzae, which belongs to the glycoside hydrolase family 78, was cloned and expressed in pichia pastoris. sds-page of the purified recombinant α-l-rhamnosidase protein revealed smeared bands with apparent molecular mass of 90-130 kda. after n-deglycosylation, the recombinant enzyme showed a molecular mass of 70 kda. the enzyme exhibited optimal activity at a ph of 5.0 and a temperature of 70 °c. specific activity of the enzyme was higher toward h ...201728800907
characteristics of korean alcoholic beverages produced by using rice nuruks containing aspergillus oryzae n159-1.herein, nuruks derived from non-glutinous and glutinous rice inoculated with aspergillus oryzae n159-1 (having high alpha-amylase and beta-glucosidase activities) were used to produce korean alcoholic beverages. the resultant beverages had enhanced fruity (ethyl caproate and isoamyl alcohol) and rose (2-phenethyl acetate and phenethyl alcohol) flavors and high taste scores.201728781547
synthesis of supermacroporous cryogel for bioreactors continuous starch hydrolysis.a bioreactor was built by means of immobilizing alpha-amylase from aspergillus oryzae by encapsulation, through cryopolymerization of acrylamide monomers for the continuous starch hydrolysis. the starch hydrolysis was evaluated regarding ph, the concentration of immobilized amylase on cryogel, the concentration of starch solution and temperature. the maximum value for starch hydrolysis was achieved at ph 5.0, concentration of immobilized enzyme 111.44mg amylase /gcriogel , concentration of starc ...201728777449
identification and functional analysis of a new glyphosate resistance gene from a fungus cdna library.glyphosate is a widely used broad spectrum herbicide; however, this limits its use once crops are planted. if glyphosate-resistant crops are grown, glyphosate can be used for weed control in crops. while several glyphosate resistance genes are used in commercial glyphosate tolerant crops, there is interest in identifying additional genes for glyphosate tolerance. this research constructed a high-quality cdna library form the glyphosate-resistant fungus aspergillus oryzae rib40 to identify genes ...201728755696
purification and characterization of a protease-resistant phytase of aspergillus oryzae sbs50 whose properties make it exceptionally useful as a feed supplement.an extracellular phytase of aspergillus oryzae sbs50 was purified to homogeneity using ammonium sulphate precipitation, ion-exchange and gel filtration chromatography. purified phytase has a monomeric molecular mass of ∼80kda exhibiting its optimal activity at ph 5.0 and 50°c with a t 1/2 of 300min at 50°c. phytase of a. oryzae displayed broad substrate specificity with vmax and km values of 58.82μmol/ml/min and 1.14mm, respectively, for calcium phytate. purity and homogeneity of the phytase was ...201728527994
impacts of amino acid substitutions in fungal cytochrome p450 monooxygenase (cyp57b3) on the effective production of 3΄-hydroxygenistein.aspergillus oryzae cytochrome p450 monooxygenase (cyp57b3) is capable of catalyzing hydroxylation of genistein to produce 3΄-hydroxygenistein. because hydroxylated derivatives of genistein, including 3΄-hydroxygenistein, exhibit various pharmacological activities, cyp57b3 would potentially be useful as a biocatalyst in the pharmaceutical field. we therefore performed random mutagenesis of cyp57b3 to improve its catalytic activities for genistein. random mutations were introduced by error-prone p ...201728520936
asp30 of aspergillus oryzae cutinase cutl1 is involved in the ionic interaction with fungal hydrophobin rola.aspergillus oryzae hydrophobin rola adheres to the biodegradable polyester polybutylene succinate-co-adipate (pbsa) and promotes pbsa degradation by interacting with a. oryzae polyesterase cutl1 and recruiting it to the pbsa surface. in our previous studies, we found that positively charged amino acid residues (h32, k34) of rola and negatively charged residues (e31, d142, d171) of cutl1 are important for the cooperative ionic interaction between rola and cutl1, but some other charged residues in ...201728475418
mitogenic lectins from cephalosporium curvulum (csl) and aspergillus oryzae (aol) mediate host-pathogen interactions leading to mycotic keratitis.a core-fucose-specific lectin, csl from cephalosporium curvulum, has been reported earlier. here we assign the role for csl and another lectin aol, from pathogenic fungus aspergillus oryzae, in causing mycotic keratitis. csl and aol show strong binding to immortalized and primary human corneal epithelial cells (hcecs) which are inhibited by asialofetuin, confirming their glycan-mediated binding. csl and aol showed increase in viability at lower concentrations (0.07 µg/ml) whereas at higher conce ...201728470344
expression and activity analysis of fructosyltransferase from aspergillus oryzae.the fructosyltransferase gene was isolated and cloned from aspergillus oryzae. the gene was 1368 bp, which encoded a protein of 455 amino acids. to analyze the activity of the expressed fructosyltransferase, the pet32a-fructosyltransferase recombined plasmid was transformed into escherichia coli bl21. the fructosyltransferase gene was successfully expressed by isopropyl-β-d-thiogalactoside (iptg) induction. the molecular weight of the expression protein was about 45 kda. the optimal conditions o ...201728639160
pathway and rate-limiting step of glyphosate degradation by aspergillus oryzae a-f02.aspergillus oryzae a-f02, a glyphosate-degrading fungus, was isolated from an aeration tank in a pesticide factory. the pathway and rate-limiting step of glyphosate (gp) degradation were investigated through metabolite analysis. gp, aminomethylphosphonic acid (ampa), and methylamine were detected in the fermentation liquid of a. oryzae a-f02, whereas sarcosine and glycine were not. the pathway of gp degradation in a. oryzae a-f02 was revealed: gp was first degraded into ampa, which was then degr ...201728636478
comparison of fungicidal properties of non-thermal plasma produced by corona discharge and dielectric barrier discharge.the inactivation of four micromycete species by action of non-thermal plasma was followed. two sources of plasma were compared, namely, positive corona discharge and dielectric barrier discharge. the corona discharge appeared as suitable for fungal spore inactivation in water suspension, whereas the barrier discharge inactivated spores on the surface of cultivation agar. cladosporium sphaerospermum was the most sensitive, being inactivated within 10 min of exposure to plasma, whereas aspergillus ...201728623536
design, synthesis, and biological evaluation of vanillin hydroxamic acid derivatives as novel peptide deformylase inhibitors.infectious disease is increasingly hampering human health, which challenge the discovery of new antibacterial target. peptide deformylase (pdf), a metalloenzyme responsible for catalyzing the remove of the n-formyl group from nascent proteins, was considered as an important target in antibacterial drug discovery.201728606047
improving the temperature characteristics and catalytic efficiency of a mesophilic xylanase from aspergillus oryzae, aoxyn11a, by iterative mutagenesis based on in silico design.to improve the temperature characteristics and catalytic efficiency of a glycoside hydrolase family (ghf) 11 xylanase from aspergillus oryzae (aoxyn11a), its variants were predicted based on in silico design. firstly, gly(21) with the maximum b-factor value, which was confirmed by molecular dynamics (md) simulation on the three-dimensional structure of aoxyn11a, was subjected to site-saturation mutagenesis. thus, one variant with the highest thermostability, aoxyn11a(g21i), was selected from the ...201728508385
rewiring the reductive tricarboxylic acid pathway and l-malate transport pathway of aspergillus oryzae for overproduction of l-malate.aspergillus oryzae finds wide application in the food, feed, and wine industries, and is an excellent cell factory platform for production of organic acids. in this work, we achieved the overproduction of l-malate by rewiring the reductive tricarboxylic acid (rtca) pathway and l-malate transport pathway of a. oryzae nrrl 3488. first, overexpression of native pyruvate carboxylase and malate dehydrogenase in the rtca pathway improved the l-malate titer from 26.1gl(-1) to 42.3gl(-1) in shake flask ...201728506930
comprehensive analysis of fungal diversity and enzyme activity in nuruk, a korean fermenting starter, for acquiring useful fungi.nuruk is a fermenting starter that is involved in the production of alcoholic beverages, and has been used in south korea for a very long time. to analyze the fungal diversity, we collected a total of 59 nuruk samples from several companies and persons in 2013 to 2014, and obtained 364 isolates. all of the single isolated fungi were identified, both morphologically and molecularly, based on the sequences of ribosomal rna gene [18s, its1-5.8s-its2, and 26s (d1/d2 region)]. in 46 nuruk samples out ...201728455587
improved α-amylase production by dephosphorylation mutation of cred, an arrestin-like protein required for glucose-induced endocytosis of maltose permease and carbon catabolite derepression in aspergillus oryzae.aspergillusoryzae produces copious amount of amylolytic enzymes, and malp, a major maltose permease, is required for the expression of amylase-encoding genes. the expression of these genes is strongly repressed by carbon catabolite repression (ccr) in the presence of glucose. malp is transported from the plasma membrane to the vacuole by endocytosis, which requires the homolog of e6-ap carboxyl terminus ubiquitin ligase hula, an ortholog of yeast rsp5. in yeast, arrestin-like proteins mediate en ...201728455339
dietary safety of a dual-enzyme preparation for animal feed: acute and subchronic oral toxicity and genotoxicity studies.animal feed is routinely supplemented with exogenous enzymes to improve nutrient utilization, such as proteases to enhance protein hydrolysis in vivo and xylanases to alleviate feed related anti-nutritional factors. the present studies were conducted to evaluate the potential oral toxicity and genotoxicity of a dual-enzyme preparation, vegpro(®) concentrate (vpr-c). acute oral toxicity studies were conducted in adult male and female sprague-dawley crl cd rats and chs swiss ico:ofi (iops caw) mic ...201728583667
combinatorial biosynthesis of (+)-daurichromenic acid and its halogenated analogue.daurichromenic acid is a meroterpenoid with various pharmacological activities that is biosynthesized from grifolic acid in rhododendron dauricum. heterologous expression of grifolic acid synthases from stachybotrys bisbyi and a daurichromenic acid synthase from r. dauricum in aspergillus oryzae mediated three-step combinatorial biosynthesis of (+)-daurichromenic acid through enantioselective 6-endo-trig cyclization. additional introduction of a halogenase from fusarium sp. into the strain resul ...201728541042
isotope-coded derivatization based lc/esi-ms/ms methods using a pair of novel reagents for quantification of hydroxycinnamic acids and hydroxybenzoic acids in fermented brown rice product.hydroxycinnamic acids (hcas) and hydroxybenzoic acids (hbas) are antioxidant phytochemicals found in rice and effective for the prevention of human diseases including cancer. fbra, which is a functional food manufactured by fermenting brown rice and rice bran with aspergillus oryzae, has been demonstrated to have chemopreventive effects against carcinogenesis in various organs. in this study, we developed methods for the relative and absolute quantification of ferulic acid, sinapic acid, caffeic ...201728531830
improved production of kojic acid by mutagenesis of aspergillus flavus hak1 and aspergillus oryzae hak2 and their potential antioxidant activity.two wild-type (wt) aspergillus strains, a. flavus hak1 and a. oryzae hak2, were selected for kojic acid (ka) biosynthesis. malt extract sucrose culture medium (mes) was the best culture medium for maximum production of ka. the maximum production of ka has been estimated at ph 4 after 7 days of incubation at 30 °c. overproduction of ka was attained by mutagenesis of both a. flavus hak1 and a. oryzae hak2 through their exposer to different doses of gamma irradiation. the mutant strains (mt) a. fla ...201728794931
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