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high-level expression of lactobacillus beta-galactosidases in lactococcus lactis using the food-grade, nisin-controlled expression system nice.in this work the overlapping genes (lacl and lacm) encoding heterodimeric beta-galactosidases from lactobacillus reuteri , lb. acidophilus , lb. sakei , and lb. plantarum were cloned into two different nisin-controlled expression (nice) vectors and expressed using lactococcus lactis nz9000 and nz3900 as hosts. the lacl gene, encoding the large subunit of the beta-galactosidases, was fused translationally downstream of the nisin-inducible promoter nisa. chloramphenicol was employed as selection m ...201020092320
enhanced thermal and ultrasonic stability of a fungal protease encapsulated within biomimetically generated silicate nanospheres.dendrimers are highly branched synthetic macromolecules with a globular shape. they have been successfully used for generation of nanospheres at mild conditions via biomimetic silicification. encapsulation of enzyme molecules within these nanospheres during their synthesis is a promising method for rapid and efficient entrapment of several enzymes. however, encapsulation of proteolytic enzymes has been rarely done via biomimetic silicification. as well, the operational stability of encapsulated ...201020100547
molecular cloning, characterization and expression analysis of two beta-n-acetylhexosaminidase homologs of coccidioides posadasii.two full-length cdnas were isolated from coccidioides posadasii that encode two deduced proteins (cphex1 and cphex2) with homology to the glycosyl hydrolase 20 family of beta-n-acetylhexosaminidases. cphex1 consists of 595 amino acids, has a predicted molecular mass of 68 kda and shares the highest identity with the n-acetylhexosaminidase (naga) of aspergillus nidulans, while cphex2 consists of 603 amino acids, has a predicted molecular mass of 68.5 kda and shares the highest identity with nag1 ...201020109094
molecular strategy for identification in aspergillus section flavi.abstract aspergillus flavus is one of the most common contaminants that produces aflatoxins in foodstuffs. it is also a human allergen and a pathogen of animals and plants. aspergillus flavus is included in the aspergillus section flavi that comprises 11 closely related species producing different profiles of secondary metabolites. a six-step strategy has been developed that allows identification of nine of the 11 species. first, three real-time pcr reactions allowed us to discriminate four grou ...201020113354
l-ascorbate, a strong inducer of l-dopa (3,4-dihydroxy-l-phenylalanine) production from pre-grown mycelia of aspergillus oryzae nrrl-1560.the inductive effect of l-ascorbate on the microbiological production of l-dopa (3,4-dihydroxy-l-phenylalanine) from aspergillus oryzae nrrl-1560 was investigated. all biochemical reactions were performed aerobically using mould mycelia as a source of enzyme tyrosinase and acetate buffer (ph 3.0) as an extractant. l-tyrosine as a substrate was added at a level of 2.5 mg/ml. maximal l-dopa production (1.876 mg/ml) was achieved when l-ascorbate (5.0 mg/ml) was added 6 min after the initiation of t ...201020121699
characterization of aspergillus oryzae glycoside hydrolase family 43 beta-xylosidase expressed in escherichia coli.this is the first report of glycoside hydrolase family 43 beta-xylosidase from aspergillus oryzae. to characterize this enzyme, the recombinant enzyme was expressed in escherichia coli. unlike known beta-xylosidases from fungal origins, the enzyme did not show substrate ambiguity and was stable at alkaline ph.201020129093
analysis of wide-domain transcriptional regulation in solid-state cultures of aspergillus oryzae.many filamentous fungi secrete considerable quantities of enzymes including protease, cellulase and xylanase, which are of major industrial importance. over the past few decades, many of these fungal enzymes have been isolated and their relevant genes characterised. solid-state fermentation (ssf), an ancient technique described as a fermentation process performed on non-soluble material whereby the material acts as a physical support and as a source of nutrients, is widely employed in the produc ...201020145973
production of xylanase with a transformant of aspergillus oryzae rib40 in a liquid-surface immobilization (lsi) system.xylanase production by a xynf1 (33 kda)-transformant of aspergillus oryzae rib40 was compared between submerged cultivation (smc) and liquid-surface immobilization (lsi) systems. while the accumulation of xylanase in the smc decreased by prolonged incubation, lsi system enabled the continuation of xylanase production to afford 4.5-fold xylanase production compared with the smc system.201020159567
cultivation characteristics and gene expression profiles of aspergillus oryzae by membrane-surface liquid culture, shaking-flask culture, and agar-plate culture.we cultivated a filamentous fungus, aspergillus oryzae iam 2706 by three different cultivation methods, i.e., shaking-flask culture (sfc), agar-plate culture (apc), and membrane-surface liquid culture (mslc), to elucidate the differences of its behaviors by different cultivation methods under the same media, by measuring the growth, secretion of proteases and alpha-amylase, secreted protein level, and gene transcriptional profile by the dna microarray analysis. the protease activities detected b ...201020159576
what does genetic diversity of aspergillus flavus tell us about aspergillus oryzae?aspergillus flavus and aspergillus oryzae belong to aspergillus section flavi. they are closely related and are of significant economic importance. the former species has the ability to produce harmful aflatoxins while the latter is widely used in food fermentation and industrial enzyme production. this review summarizes the current understanding of the similarity of the a. flavus and a. oryzae genomes, the genetic diversity in a. flavus and a. oryzae populations, the causes of this diversity, a ...201020163884
characterization of water-soluble complexes of polyacrylic acid with alpha-amylase from aspergillus oryzae.polycomplex formation of alpha-amylase from aspergillus oryzae (taka) with polyacrylic acid (paa) was studied by ph titration, fluorescence, and high performance liquid chromatography (hplc) methods in water solutions. according to the our results, the complex formation and its solubility were depended on nature of enzyme and the ph of solutions. both of them correlates isoelectric points (pi). the stability of paa-amylase complexes was negligibly weak at ph 7 [ph > pi (isoelectric ph)]. stable ...201020165908
promotion of efficient saccharification of crystalline cellulose by aspergillus fumigatus swo1.swollenin is a protein from trichoderma reesei that has a unique activity for disrupting cellulosic materials, and it has sequence similarity to expansins, plant cell wall proteins that have a loosening effect that leads to cell wall enlargement. in this study we cloned a gene encoding a swollenin-like protein, swo1, from the filamentous fungus aspergillus fumigatus, and designated the gene afswo1. afswo1 has a bimodular structure composed of a carbohydrate-binding module family 1 (cbm1) domain ...201020173066
aspergillus oryzae flavohemoglobins promote oxidative damage by hydrogen peroxide.apart from their well-established role in nitric oxide detoxification, flavohemoglobins (fhbs) are also believed to be involved in protection against oxidative stress in some yeast and bacteria. however, different studies have reported contradictory results in this regard. here, we investigate the relationship between two fhbs in aspergillus oryzae (cytosolic fhb1 and mitochondrial fhb2) and oxidative stress. the strains deficient in the two fhbs exhibited higher resistance to hydrogen peroxide ...201020211603
some biochemical and physical changes during the preparation of the enzyme-ripening sufu, a fermented product of soybean curd.in this study, sufu, a fermented product of soybean curd, was prepared by ripening the salted tofu in the mash of aspergillus oryzae -fermented rice-soybean koji possessing various hydrolytic enzymes for 16 days. it was found that protease, alpha-amylase, beta-amylase, and lipase activities observed in the koji granules reduced most pronouncedly during the initial 4 or 8 days of ripening. meanwhile, an increase in the activity of the various enzymes was noted in the ripening infusion and tofu cu ...201020218723
stimulation of lignocellulosic biomass hydrolysis by proteins of glycoside hydrolase family 61: structure and function of a large, enigmatic family.currently, the relatively high cost of enzymes such as glycoside hydrolases that catalyze cellulose hydrolysis represents a barrier to commercialization of a biorefinery capable of producing renewable transportable fuels such as ethanol from abundant lignocellulosic biomass. among the many families of glycoside hydrolases that catalyze cellulose and hemicellulose hydrolysis, few are more enigmatic than family 61 (gh61), originally classified based on measurement of very weak endo-1,4-beta-d-gluc ...201020230050
mutation of key residues in the c-methyltransferase domain of a fungal highly reducing polyketide synthase.site directed mutations of the c-methyltransferase domain of squalestatin tetraketide synthase were made in an attempt to alter the methylation pattern of the synthase expressed in vivo: mutation resulted in either no effect or in complete abrogation of polyketide production.201020237645
a study of the action of purified amylase from aspergillus oryzae, taka-amylase. 194720272106
the formation of kojic acid (3-hydroxy-6-hydroxymethyl-gamma pyrone) from ethyl alcohol by aspergillus oryzae. 194620277276
development and refinement of a high-efficiency gene-targeting system for aspergillus flavus.an efficient gene-targeting system based on impairment of the nonhomologous end-joining pathway and the orotidine monophosphate decarboxylase gene (pyrg) in aspergillus flavus was established. it was achieved by replacing the ku70 gene with the aspergillus oryzae pyrithiamine resistance (ptr) gene and by inserting the aspergillus parasiticus cypa gene into the pyrg locus. the utility of this system was demonstrated by disruption of nine candidate genes for conidial pigment biosynthesis. the gene ...201020298723
analysis of genome-wide coexpression and coevolution of aspergillus oryzae and aspergillus niger.analysis of coexpressed genes in response to different perturbations at the genome-level can provide new insight into global regulatory structures. here we performed integrated data analysis for a crossspecies comparative investigation by exploring genomes and transcriptional coexpression profiles in aspergillus oryzae and aspergillus niger. based on our analysis of conserved coexpressed genes, fatty acid catabolism via beta-oxidation, fatty acid transport, the glyoxylate bypass, and peroxisomal ...201020337533
molecular characterization and isolation of cytochrome p450 genes from the filamentous fungus aspergillus oryzae.we explored the molecular diversity of cytochrome p450 genes in the filamentous fungus aspergillus oryzae using bioinformatic and experimental approaches. based on bioinformatic annotation, we found 155 putative genes of cytochromes p450 in the whole genome sequence; however, 13 of 155 appeared to be pseudogenes due to sequence deletions and/or inframe stop codon(s). there are 87 families of a. oryzae cytochromes p450 (aocyps), indicating considerable phylogenetic diversity. to characterize a. o ...201020358180
molecular strategy for identification in aspergillus section flavi.aspergillus flavus is one of the most common contaminants that produces aflatoxins in foodstuffs. it is also a human allergen and a pathogen of animals and plants. aspergillus flavus is included in the aspergillus section flavi that comprises 11 closely related species producing different profiles of secondary metabolites. a six-step strategy has been developed that allows identification of nine of the 11 species. first, three real-time pcr reactions allowed us to discriminate four groups within ...201020377644
survey of the transcriptome of aspergillus oryzae via massively parallel mrna sequencing.aspergillus oryzae, an important filamentous fungus used in food fermentation and the enzyme industry, has been shown through genome sequencing and various other tools to have prominent features in its genomic composition. however, the functional complexity of the a. oryzae transcriptome has not yet been fully elucidated. here, we applied direct high-throughput paired-end rna-sequencing (rna-seq) to the transcriptome of a. oryzae under four different culture conditions. with the high resolution ...201020392818
effect of halotolerant starter microorganisms on chemical characteristics of fermented chum salmon (oncorhynchus keta) sauce.chum salmon sauce mash was inoculated with barley koji (barley steamed and molded with aspergillus oryzae ) and halotolerant microorganisms (htms), zygosaccharomyces rouxii , candida versatilis , and tetragenococcus halophilus , in nine different combinations under non-aseptic conditions similar to the industrial fish sauce production and fermented at 35 +/- 2.5 degrees c for 84 days. the changes in the chemical components, color, and sensory properties during fermentation were investigated. fre ...201020405947
application of exogenous enzymes to beef muscle of high and low-connective tissue.exogenous enzymes tenderize meat through proteolysis. triceps brachii and supraspinatus were randomly assigned to the seven enzyme treatments, papain, ficin, bromelain, homogenized fresh ginger, bacillus subtilis protease, and two aspergillus oryzae proteases or control to determine the extent of tenderization (warner-bratzler shear and sensory evaluation) and mode of action (myofibrillar or collagen degradation). sensory evaluation showed improvement (p<0.0009) for tenderness and connective tis ...201020416788
microbiota during fermentation of chum salmon (oncorhynchus keta) sauce mash inoculated with halotolerant microbial starters: analyses using the plate count method and pcr-denaturing gradient gel electrophoresis (dgge).nine different combinations of mugi koji (barley steamed and molded with aspergillus oryzae) and halotolerant microorganisms (htms), zygosaccharomyces rouxii, candida versatilis, and tetragenococcus halophilus, were inoculated into chum salmon sauce mash under a non-aseptic condition used in industrial fish sauce production and fermented at 35 +/- 2.5 degrees c for 84 days to elucidate the microbial dynamics (i.e., microbial count and microbiota) during fermentation. the viable count of halotole ...201020417400
direct microbial conversion of wheat straw into lipid by a cellulolytic fungus of aspergillus oryzae a-4 in solid-state fermentation.direct microbial conversion of wheat straw into lipid by a cellulolytic fungus of aspergillus oryzae a-4 in solid-state fermentation (ssf) was investigated. in submerged fermentation, a. oryzae a-4 accumulated lipid to 15-18.15% of biomass when pure cellulose was utilized as the sole substrate. in ssf of the wheat straw and bran mixture, a. oryzae a-4 yielded lipid of 36.6mg/g dry substrate (gds), and a cellulase activity of 1.82 fpu/gds with 25.25% of holocellulose utilization in the substrates ...201020444596
high-level expression and efficient purification of bioactive swollenin in aspergillus oryzae.the bioactivity of swollenin is beneficial to cellulose decomposition by cellulase despite the lack of hydrolytic activity itself. in order to improve the productivity of swollenin, the effects of culture conditions on the expression level in recombinant aspergillus oryzae were investigated systematically. with regard to the bioactivity of swollenin, glycerin and peanut meal were the optimal carbon or nitrogen source, respectively. the highest level production of swollenin (50 mg l(-1)) was atta ...201020446057
functional hypothesis on miraculin' sweetness by a molecular dynamics approach.miraculin differs from other sweet-tasting proteins because it is a taste-modifier having the unusual property of modifying sourness into sweetness. its dimer is covalently linked by an inter-chain disulphide bond, and shows its taste-modifying activity at acidic ph, with maximum at ph 3.0, while it is flat at neutral ph. previous studies suggested the importance of two histidine residues for the taste-modifying activity of miraculin. in this work, we have conducted molecular dynamics simulation ...201020451498
a eukaryotic copper-containing nitrite reductase derived from a nirk homolog gene of aspergillus oryzae.we cloned a bacterial copper-containing nitrite reductase (nirk) homolog gene of aspergillus oryzae (aonirk). alignment showed that amino acid residues crucial for copper binding are conserved in the deduced sequence of the fungal protein. the recombinant protein exhibited distinct nitrite reductase activity, and its absorption and epr spectra showed the presence of type 1 and type 2 copper atoms in the molecule. aonirk transcriptionally responded to denitrification conditions. although the deni ...201020460712
molecular cloning of ocpo encoding carboxypeptidase o of aspergillus oryzae iam2640.carboxypeptidase o from aspergillus oryzae iam2640 is a serine-type carboxypeptidase. in this study, we cloned and sequenced cdna and genomic dna carrying ocpo encoding carboxypeptidase o. the results showed that the length of ocpo was 1,816 bp, and the open reading frame encoded a putative preproenzyme composed of 472 amino acid residues of the mature carboxypeptidase o and an additional n-terminal sequence of 50 amino acid residues. a blastn search revealed that a gene, ao090020000351, in a. o ...201020460731
use of the aspergillus oryzae actin gene promoter in a novel reporter system for exploring antifungal compounds and their target genes.demand for novel antifungal drugs for medical and agricultural uses has been increasing because of the diversity of pathogenic fungi and the emergence of drug-resistant strains. genomic resources for various living species, including pathogenic fungi, can be utilized to develop novel and effective antifungal compounds. we used aspergillus oryzae as a model to construct a reporter system for exploring novel antifungal compounds and their target genes. the comprehensive gene expression analysis sh ...201020464390
4-deoxy-substrates for beta-n-acetylhexosaminidases: how to make use of their loose specificity.beta-n-acetylhexosaminidases feature so-called wobbling specificity, which means that they cleave substrates both in gluco- and galacto- configurations, with the activity ratio depending on the enzyme source. here we present the new finding that fungal beta-n-acetylhexosaminidases are able to hydrolyze and transfer 4-deoxy-n-acetylhexosaminides with high yields. this clearly demonstrates that the 4-hydroxy moiety at the substrate pyranose ring is not essential for substrate binding to the enzyme ...201020466648
identification of csypyrone b1 as the novel product of aspergillus oryzae type iii polyketide synthase csyb.as a novel superfamily of type iii polyketide synthases (pkss) in microbes, four genes, csya, csyb, csyc, and csyd, were found in the genome of aspergillus oryzae, an industrially important filamentous fungus. although orthologs of csya, csyc, and csyd genes are present in a closely related species, aspergillus flavus, csyb gene is unique to a. oryzae. to identify its function, we carried out overexpression of csyb gene under the control of alpha-amylase promoter in a. oryzae. 3-(3-acetyl-4-hydr ...201020471846
simultaneous c7- and n1-prenylation of cyclo-l-trp-l-trp catalyzed by a prenyltransferase from aspergillus oryzae.a putative prenyltransferase gene ctrppt was amplified from aspergillus oryzae dsm1147, cloned into pqe70 and overexpressed in escherichia coli. the overproduced his(6)-ctrppt was purified to near homogeneity and incubated with l-tryptophan or tryptophan-containing cyclic dipeptides in the presence of dimethylallyl diphosphate. the formation of the enzyme products was monitored with hplc. it was shown that ctrppt differed clearly from other known indole prenyltransferases in several aspects. thi ...201020473424
antimicrobial and antioxidant activities of the essential oil from herba moslae.herba moslae is a well-known edible and medicinal plant in china, and the essential oil, which is assumed to contain the active components, was isolated by steam distillation method. the aim of this study was to investigate the antimicrobial activity and the antioxidant activity of the essential oil from herba moslae.201020474054
solanapyrone synthase, a possible diels-alderase and iterative type i polyketide synthase encoded in a biosynthetic gene cluster from alternaria solani.the solanapyrone biosynthetic gene cluster was cloned from alternaria solani. it consists of six genes-sol1-6-coding for a polyketide synthase, an o-methyltransferase, a dehydrogenase, a transcription factor, a flavin-dependent oxidase, and cytochrome p450. the prosolanapyrone synthase (pss) encoded by sol1 was expressed in aspergillus oryzae and its product was identified as desmethylprosolanapyrone i (8). although pss is closely related to the pkss/diels-alderases lovb and mlca of lovastatin a ...201020486243
display of both n- and c-terminal target fusion proteins on the aspergillus oryzae cell surface using a chitin-binding module.a novel cell surface display system in aspergillus oryzae was established by using a chitin-binding module (cbm) from saccharomyces cerevisiae as an anchor protein. cbm was fused to the n or c terminus of green fluorescent protein (gfp) and the fusion proteins (gfp-cbm and cbm-gfp) were expressed using a. oryzae as a host. western blotting and fluorescence microscopy analysis showed that both gfp-cbm and cbm-gfp were successfully expressed on the cell surface. in addition, cell surface display o ...201020499230
a randomized controlled trial to assess the impact of dietary energy sources, feed supplements, and the presence of super-shedders on the detection of escherichia coli o157:h7 in feedlot cattle using different diagnostic procedures.alteration of the gastro-intestinal tract through manipulation of cattle diets has been proposed as a preharvest control measure to reduce fecal shedding of escherichia coli o157:h7. the objective of this study was to examine the effects of the energy source's moisture content (high moisture corn and dry whole-shelled corn), two natural feed supplements (saccaromyces cerevisiae boulardii cncm 1079-levucell and aspergillus oryzae-amaferm), and two levels of vitamin a (2200 iu/kg and no supplement ...201020500082
uptake of glycosylated ?-galactosidase into human brain synaptosomes: occurrence of two different systems for specific glycosides.uptake of glycoproteins into brain nerve terminals was studied, using glycosylated ?-galactosidase and human brain synaptosomes. a series of p-aminophenyl ?- or ?-glycosides were covalently bound to ?-galactosidase purified from aspergillus oryzae and uptake of the enzyme into human brain synaptosomes was measured. ?-galactosidase coupled with ?-d-mannoside (?-d-man ?-gal) was the best to be taken up among glycoconjugates tested. the uptake was dependent on the amount of ?-d-man ?-gal, the incub ...198820501274
expression and export: recombinant protein production systems for aspergillus.several aspergillus species, in particular aspergillus niger and aspergillus oryzae, are widely used as protein production hosts in various biotechnological applications. in order to improve the expression and secretion of recombinant proteins in these filamentous fungi, several novel genetic engineering strategies have been developed in recent years. this review describes state-of-the-art genetic manipulation technologies used for strain improvement, as well as recent advances in designing the ...201020532762
penicillin biosynthesis in aspergillus oryzae and its overproduction by genetic engineering.aspergillus oryzae penicillin biosynthetic genes were clustered. the penicillin production was positively regulated by vea, a global gene regulator required for transcriptional expression of the penicillin biosynthetic genes. overexpression of the biosynthetic genes by a strong promoter yielded a greater than 100-fold increase in penicillin production.201020541108
alternative method for quantification of alfa-amylase activity.a modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. the proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. thus, a standard curve was built using alfa-amylase solution from aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 u.ml-1. clear radial diffusion zones were measured after 4 hours of incubation at 20 a degrees c. a linear relationship between the logarit ...201020552148
gene cloning and enzyme structure modeling of the aspergillus oryzae n74 fructosyltransferase.the fructooligosaccharides (fos) represent an important source of prebiotic compounds that are widely used as an ingredient in functional foods. recently, the strain aspergillus oryzae n74 was reported as a potential microorganism for the industrial production of fos, due to its high yields of fos production. in this work, we used a pcr-cloning strategy to clone the a. oryzae n74 ftase gene as a previous step for recombinant enzyme production. ftase showed a 1630 bp size with a 99% similarity wi ...201120563857
applicability of penicillium chrysogenum rheological correlations to broths of other fungal strains.existing correlations of power law consistency index with penicillium chrysogenum biomass concentration and morphology were revised using a microscope magnification of 50 times to characterize the latter, rather than the 80 times used previously. this allowed tests of the correlations on broths of aspergillus oryzae and aspergillus niger, which have such large mycelial sizes that a lower magnification is required for accurate morphological analysis. the new correlations were successful at predic ...201020574833
first heterologous reconstruction of a complete functional fungal biosynthetic multigene cluster. 201020575135
identifying 8-hydroxynaringenin as a suicide substrate of mushroom tyrosinase.a biotransformed metabolite of naringenin was isolated from the fermentation broth of aspergillus oryzae, fed with naringenin, and identified as 8-hydroxynaringenin based on the mass and (1)h- and (13)c-nmr spectral data. the compound showed characteristics of both an irreversible inhibitor and a substrate of mushroom tyrosinase in preincubation and hplc analysis. these results demonstrate that 8-hydroxynaringenin belongs to a suicide substrate of mushroom tyrosinase. the partition ratio between ...201020587349
a novel pathway for nicotine degradation by aspergillus oryzae 112822 isolated from tobacco leaves.an efficient nicotine-degrading fungus was isolated from tobacco leaves and identified as aspergillus oryzae 112822 based on morphological characteristics and sequence analysis of 18s rdna, 5.8s rdna and the internal transcribed spacer (5.8s-its region). when the strain was cultured in a medium with tobacco leaf extract for 40 h, the maximum amount of cell growth was 3.6 g l(-1) and nicotine degradation was 2.19 g l(-1). the intermediates of nicotine degradation by resting cells were isolated by ...201020600861
enhanced production and secretion of heterologous proteins by the filamentous fungus aspergillus oryzae via disruption of vacuolar protein sorting receptor gene aovps10.filamentous fungi have received attention as hosts for heterologous protein production because of their high secretion capability and eukaryotic posttranslational modifications. however, despite these positive attributes, a bottleneck in posttranscriptional processing limits protein yields. the vacuolar protein sorting gene vps10 encodes a sorting receptor for the recognition and delivery of several yeast vacuolar proteins. although it can also target recombinant and aberrant proteins for vacuol ...201020622126
aspergillus oryzae type iii polyketide synthase csya is involved in the biosynthesis of 3,5-dihydroxybenzoic acid.as a novel superfamily of type iii polyketide synthases in microbes, four genes csya, csyb, csyc, and csyd, were found in the genome of aspergillus oryzae, an industrially important filamentous fungus. in order to analyze their functions, we carried out the overexpression of csya under the control of alpha-amylase promoter in a. oryzae and identified 3,5-dihydroxybenzoic acid (dhba) as the major product. feeding experiments using (13)c-labeled acetates confirmed that the acetate labeling pattern ...201020630753
study on the synthesis of sucrose-6-acetate catalyzed by fructosyltransferase from aspergillus oryzae.the study had mainly investigated the synthesis of sucrose-6-acetate (s-6-a) in fructosyltransferase action. the synthesis reaction of s-6-a was performed between sucrose and glucose-6-acetate (g-6-a), and identified by high performance liquid chromatography (hplc). according to the reaction of s-6-a catalyzed by fructosyltransferase from aspergillus oryzae, the effect factors of reaction, such as the ratio of g-6-a to sucrose, temperature, time, ph, substrate and enzyme concentration in the rea ...201120643232
microbiological and biochemical survey on the transition of fermentative processes in fukuyama pot vinegar brewing.traditional brewing of fukuyama pot vinegar is a process that has been continued in fukuyama, kagoshima, japan, for almost 200 years. the entire process proceeds from raw materials, including steamed rice, rice koji (steamed rice grown with a fungus, aspergillus oryzae) and water, to produce vinegar in roughly capped large pots laid in the open air. no special fermentative manipulation is required, except for scattering dried rice koji (called furi-koji) on the surface of the mash to form a cap- ...201020647677
a novel enzymatic microreactor with aspergillus oryzae β-galactosidase immobilized on silicon dioxide nanosprings.the use of silicon dioxide (sio(2) ) nanosprings as supports for immobilized enzymes in a continuous microreactor is described. a nanospring mat (2.2 cm(2) × 60 μm thick) was functionalized with γ-aminopropyltriethoxysilane, then treated with n-succinimidyl-3-(2-pyridyldithio)-propionate (spdp) and dithiothreitol (dtt) to produce surface thiol (--sh) groups. spdp-modified β-galactosidase from aspergillus oryzae was immobilized on the thiolated nanosprings by reversible disulfide linkages. the en ...201020661927
a comparative study of activity and apparent inhibition of fungal β-glucosidases.β-glucosidases (bgs) from aspergillus fumigates, aspergillus niger, aspergillus oryzae, chaetomium globosum, emericella nidulans, magnaporthe grisea, neurospora crassa, and penicillium brasilianum were purified to homogeneity, and analyzed by isothermal titration calorimetry with respect to their hydrolytic activity and its sensitivity to glucose (product) using cellobiose as substrate. global non-linear regression of several reactions, with or without added glucose, to a product inhibition equa ...201020677177
simple metabolite extraction method for metabolic profiling of the solid-state fermentation of aspergillus oryzae.solid-state culture of microorganisms is an important style of culture both in the traditional food industry and in the modern fermentation industry. we propose here a simple method for metabolite extraction from the solid-state fermentation of a filamentous fungus, aspergillus oryzae, which is known as rice-koji. to evaluate the efficiency of metabolite extraction, liquid chromatography-mass spectrometry (lc/ms) was used for simultaneous detection of a wide range of metabolites including amino ...201020685162
enzymatic synthesis and characterization of hydroquinone galactoside using kluyveromyces lactis lactase.hydroquinone galactoside (hq-gal) as a potential skin whitening agent was synthesized by the reaction of lactase (beta-galactosidase) from kluyveromyces lactis, aspergillus oryzae, bacillus circulans, and thermus sp. with lactose as a donor and hq as an acceptor. among these lactases, the acceptor reaction involving hq and lactose with k. lactis lactase showed a higher conversion ratio to hq-gal (60.27%). hq-gal was purified using butanol partitioning and silica gel column chromatography. the st ...201020687552
purification and characterization of termite endogenous beta-1,4-endoglucanases produced in aspergillus oryzae.although termites are known to have a highly efficient lignocellulose-digesting system, mass production of native endogenous cellulases of termites has failed in escherchia coli, and in saccharomyces cerevisiae, and it has not been accomplished. here we report the successful production, purification, and characterization of two termite endogenous beta-1,4-endoglucanases, rseg and nteg, from the salivary gland of reticulitermes speratus and the midgut of nasutitermes takasagoensis respectively, u ...201020699551
characterization of a cellobiohydrolase (mocel6a) produced by magnaporthe oryzae.three gh-6 family cellobiohydrolases are expected in the genome of magnaporthe grisea based on the complete genome sequence. here, we demonstrate the properties, kinetics, and substrate specificities of a magnaporthe oryzae gh-6 family cellobiohydrolase (mocel6a). in addition, the effect of cellobiose on mocel6a activity was also investigated. mocel6a contiguously fused to a histidine tag was overexpressed in m. oryzae and purified by affinity chromatography. mocel6a showed higher hydrolytic act ...201020709852
three-dimensional phase-contrast imaging by a computed-tomography microscope.we report a technique for measuring the three-dimensional variation of refractive indices in a microscopic sample. the technique is an adaptation of optical computed tomography and is effective in measuring the three-dimensional refractive-index distribution of a nonabsorbing microscopic sample. our report also includes a discussion of the conditions for the unambiguous application of the technique as well as results of experiments conducted with aspergillus oryzae (commonly known as green mold) ...199220720664
secreted fungal sulfhydryl oxidases: sequence analysis and characterisation of a representative flavin-dependent enzyme from aspergillus oryzae.sulfhydryl oxidases are flavin-dependent enzymes that catalyse the formation of de novo disulfide bonds from free thiol groups, with the reduction of molecular oxygen to hydrogen peroxide. sulfhydryl oxidases have been investigated in the food industry to remove the burnt flavour of ultraheat-treated milk and are currently studied as potential crosslinking enzymes, aiming at strengthening wheat dough and improving the overall bread quality.201020727152
contribution of ethanol-tolerant xylanase g2 from aspergillus oryzae on japanese sake brewing.we purified three xylanase isozymes (xynf1, xynf3 and xyng2) from a solid-state aspergillus oryzae rib128 culture using chromatography. the results of our sake-brewing experiment, in which we used exogenously supplemented enzymes, revealed that only xyng2 improved the alcohol yield and the material utilization. the alcohol yield of the xyng2 batch displayed an increase of 4.4% in comparison to the control, and the amount of sake cake decreased by 4.6%. the contribution of xyng2 was further confi ...201020727822
a novel aspergillus oryzae esterase that hydrolyzes 4-hydroxybenzoic acid esters.in this study we report the biochemical characterization of a hypothetical protein from aspergillus oryzae exhibiting sequence identity with feruloyl esterase and tannase from the genus aspergillus. the purified recombinant protein showed a hydrolytic activity toward the ethyl, propyl, or butyl esters of 4-hydroxybenzoic acid, but did not show feruloyl esterase or tannase activity. finally, the enzyme decreased the antimicrobial activity of parabens against a. oryzae via hydrolysis of the ester ...201020728445
indoloditerpenes from an algicolous isolate of aspergillus oryzae.two new indoloditerpene derivatives asporyzin a (1) and asporyzin b (2), one new indoloditerpene asporyzin c (3), and three known related indoloditerpenes jbir-03 (4), emindole sb (5), and emeniveol (6) were isolated from an endophytic fungus aspergillus oryzae, isolated from the marine red alga heterosiphonia japonica. their structures were unambiguously established by spectroscopic techniques. in addition, all the isolates were evaluated preliminarily for insecticidal and antimicrobial activit ...201020797856
overexpression and characterization of an extracellular leucine aminopeptidase from aspergillus oryzae.leucine aminopeptidase (lap), an enzyme used in the food industry, is an exopeptidase that removes an amino acid residue, primarily leucine (leu), from the n-terminus of peptides and protein substrates. in this study, we focused on the leucine aminopeptidase a (lapa) gene from aspergillus oryzae rib40. to purify and characterize the lapa, lapa was overexpressed in a. oryzae rib40 using the amyb promoter. lap activity in the culture supernatant of one transformant harboring the lapa expression pl ...201120803144
functional analysis and subcellular location of two flavohemoglobins from aspergillus oryzae.multiple flavohemoglobin (fhb) homolog genes are found in the genomes of eukaryotic microorganisms, but their functions remain unknown. in this study, two distinct types of fhbs (predictive cytosolic fhb1 and predictive mitochondrial fhb2) from the fungus aspergillus oryzae were investigated to elucidate the physiological roles of these fhbs. the fhb1 gene responded to external nitric oxide (no) stress at the transcriptional level, whereas the fhb2 gene did not. disrupting fhb1 increased cell hy ...201120817113
crystallization and preliminary x-ray analysis of formate oxidase, an enzyme of the glucose-methanol-choline oxidoreductase family.formate oxidase (fod), which catalyzes the oxidation of formate to yield carbon dioxide and hydrogen peroxide, belongs to the glucose-methanol-choline oxidoreductase (gmco) family. fod from aspergillus oryzae rib40, which has a modified fad as a cofactor, was crystallized at 293 k by the hanging-drop vapour-diffusion method. the crystal was orthorhombic and belonged to space group c222(1). diffraction data were collected from a single crystal to 2.4 a resolution.201020823527
identification and characterization of genes responsible for biosynthesis of kojic acid, an industrially important compound from aspergillus oryzae.kojic acid is produced in large amounts by aspergillus oryzae as a secondary metabolite and is widely used in the cosmetic industry. glucose can be converted to kojic acid, perhaps by only a few steps, but no genes for the conversion have thus far been revealed. using a dna microarray, gene expression profiles under three pairs of conditions significantly affecting kojic acid production were compared. all genes were ranked using an index parameter reflecting both high amounts of transcription an ...201020849972
identification and characterization of a novel fermented substance produced by edible aspergillus oryzae ao-1 that inhibits dpp-iv activity.we identified a novel fermented substance (fka) produced by the edible aspergillus oryzae strain ao-1 that inhibited dipeptidyl peptidase iv (dpp-iv) (ic(50) = 3.41 mg · ml(-1)). hplc analysis of fka showed specific one metabolite (wyk-1), which inhibited dpp-iv (ic(50) = 6.98 μm). wyk-1 was identified as a tetrahydroxyisoquinoline derivative. interestingly, we examined 60 strains of a. oryzae, ao-1 was the only a. oryzae strain that produces wyk-1. this is the first report that an edible a. ory ...201120851672
enzyme stabilization and immobilization by sol-gel entrapment.while biocatalysts show tremendous potential for the industrial production of fine chemicals, their integration into large-scale processes has been slow. one of the main reasons for slow acceptance in industry is the inherent instability of the enzymes. recent developments in bioengineering have shed some light on methods of improving enzyme stability. one method that has been used for many decades, successfully to varying degrees, has been the immobilization of enzymes. to this regards, silica ...201120865388
in vivo imaging of endoplasmic reticulum and distribution of mutant α-amylase in aspergillus oryzae.properly folded proteins destined for secretion exit through a specific subdomain of the endoplasmic reticulum (er) known as transitional er (ter) sites or er exit sites (eres). while such proteins in filamentous fungi localize at the hyphal tips overlapping the spitzenkörper, the distribution of misfolded proteins remains unknown. in the present study, we analyzed the distribution of mutant protein as well as er and ter sites visualized by expression of aoclxa and aosec13 fused with fluorescent ...201020884367
disruption of ten protease genes in the filamentous fungus aspergillus oryzae highly improves production of heterologous proteins.proteolytic degradation by secreted proteases into the culture medium is one of the significant problems to be solved in heterologous protein production by filamentous fungi including aspergillus oryzae. double (tppa, and pepe) and quintuple (tppa, pepe, nptb, dppiv, and dppv) disruption of protease genes enhanced human lysozyme (hly) and bovine chymosin (chy) production by a. oryzae. in this study, we used a quintuple protease gene disruptant and performed successive rounds of disruption for fi ...201020957357
phylogeny of fungal hemoglobins and expression analysis of the aspergillus oryzae flavohemoglobin gene fhba during hyphal growth.the fhba genes encoding putative flavohemoglobins (fhb) from aspergillus niger and aspergillus oryzae were isolated. comparison of the deduced amino acid sequence of the a. niger fhba gene and other putative filamentous fungal fhb-encoding genes to that of ralstonia eutropha shows an overall conserved gene structure and completely conserved catalytic amino acids. several yeasts and filamentous fungi, including both aspergillus species have been found to contain a small fhb gene family mostly con ...201020960969
screening and identification of a fungal β-glucosidase and the enzymatic synthesis of gentiooligosaccharide.after screening with 0.1% esculoside and 0.03% fecl(3), we identified from rotten wood a fungal isolate hml0366 that produces high amount of β-glucosidase. phenotypic and rdna internal transcribed spacer sequence analyses indicated that the isolate belongs to aspergillus oryzae. the β-glucosidase produced by hml0366 had an activity of 128 u/g. high performance liquid chromatography analysis also demonstrated a high transglycosylation activity of the crude enzyme. the β-glucosidase was stable bet ...201020963514
adenosine deaminase from aspergillus oryzae. 194620997999
further studies of the purification and properties of the amylase of aspergillus oryzae. 194521005743
extraction and application of laccases from shimeji mushrooms (pleurotus ostreatus) residues in decolourisation of reactive dyes and a comparative study using commercial laccase from aspergillus oryzae.oxidases are able to degrade organic pollutants; however, high costs associated with biocatalysts production still hinder their use in environmental biocatalysis. our study compared the action of a commercial laccase from aspergillus oryzae and a rich extract from pleurotus ostreatus cultivation residues in decolourisation of reactive dyes: drimaren blue x-3lr (dmblr), drimaren blue x-bln (dmbbln), drimaren rubinol x-3lr (dmr), and drimaren blue c-r (rbbr). the colour removal was evaluated by co ...201021052547
heterologous expression and characterization of a glucose-stimulated β-glucosidase from the termite neotermes koshunensis in aspergillus oryzae.neotermes koshunensis is a lower termite that secretes endogenous β-glucosidase in the salivary glands. this β-glucosidase (g1nkbg) was successfully expressed in aspergillus oryzae. g1nkbg was purified to homogeneity from the culture supernatant through ammonium sulfate precipitation and anion exchange, hydrophobic, and gel filtration chromatographies with a 48-fold increase in purity. the molecular mass of the native enzyme appeared as a single band at 60 kda after gel filtration analysis, indi ...201021057947
enzyme-mediated site-specific antibody-protein modification using a zz domain as a linker.a zz domain (zz) and alkaline phosphatase (ap), luciferase (luc), or glucose oxidase (god) were conjugated using sortase a (srta) from staphylococcus aureus. the specific peptidyl linker for srta was genetically fused to the c-terminus of zz, and the other linker was fused to the n-terminus of ap, luc, or god, respectively. the resultant proteins were obtained separately by bacterial expression. the recombinant peptide-tagged zz and ap, luc, or god were site-specifically conjugated by srta throu ...201021069999
gc-ms based metabolite profiling of rice koji fermentation by various fungi.in this study, aspergillus kawachii, aspergillus oryzae, and rhizopus sp., were utilized for rice koji fermentation, and the metabolites were analyzed in a time-dependent manner by gas chromatography-mass spectrometry. on principal component analysis, the metabolite patterns were clearly distinguished based on the fungi species. this approach revealed that the quantities of glucose, galactose, and glycerol gradually increased as a function of fermentation time in all trials rice koji fermentatio ...201021071848
enzymes for the biofunctionalization of poly(ethylene terephthalate).the functionalization of synthetic polymers such as poly(ethylene terephthalate) to improve their hydrophilicity can be achieved biocatalytically using hydrolytic enzymes. a number of cutinases, lipases, and esterases active on polyethylene terephthalate have been identified and characterized. enzymes from fusarium solani, thermomyces insolens, t. lanuginosus, aspergillus oryzae, pseudomonas mendocina, and thermobifida fusca have been studied in detail. thermostable biocatalysts hydrolyzing poly ...201021076908
changes in the contents of sugars and organic acids during the ripening and storage of sufu, a traditional oriental fermented product of soybean cubes.in the present study, sufu, an oriental fermented product of soybeans, was prepared by ripening the tofu cubes in aspergillus oryzae fermented rice-soybean koji mash for 16 days at 37 °c. the sufu product was further kept at room temperature for another 30 days. examining the changes in the sugar content revealed that stachyose, raffinose, and sucrose contents of tofu and rice-soybean koji granules decreased while levels of glucose and fructose increased during the ripening period and after stor ...201021090680
breeding and identification of novel koji molds with high activity of acid protease by genome recombination between aspergillus oryzae and aspergillus niger.acid protease is essential for degradation of proteins during soy sauce fermentation. to breed more suitable koji molds with high activity of acid protease, interspecific genome recombination between a. oryzae and a. niger was performed. through stabilization with d-camphor and haploidization with benomyl, several stable fusants with higher activity of acid protease were obtained, showing different degrees of improvement in acid protease activity compared with the parental strain a. oryzae. in a ...201021107641
in situ generation of hydrogen peroxide by carbohydrate oxidase and cellobiose dehydrogenase for bleaching purposes.the carbohydrate oxidase from microdochium nivale (caox), heterologously expressed in aspergillus oryzae, and cellobiose dehydrogenase from myriococcum thermophilum (mtcdh), were assessed for their ability to generate bleaching species at a ph suitable for liquid detergents. the substrate specificities of caox and mtcdh were analyzed on a large variety of soluble and insoluble substrates, using oxygen as an electron receptor. even insoluble substrates like cellulose were oxidized from both caox ...201021117085
asporyergosterol, a new steroid from an algicolous isolate of aspergillus oryzae.asporyergosterol (1), a new steroid with an e double bond between c-17 and c-20, was identified from the culture extracts of aspergillus oryzae, an endophytic fungus isolated from the marine red alga heterosiphonia japonica. moreover, four known steroids including (22e,24r)-ergosta-4,6,8(14),22-tetraen-3-one (2), (22e,24r)-3beta-hydroxyergosta-5,8,22-trien-7-one (3), (22e,24r)-ergosta-7,22-dien-3beta,5alpha,6beta-triol (4), and (22e,24r)-5alpha,8alpha-epidioxyergosta-6,22-dien-3beta-ol (5) were ...201021121251
production of 3,4-dihydroxy l-phenylalanine by a newly isolated aspergillus niger and parameter significance analysis by plackett-burman design.the amino acid derivative 3,4-dihydroxy l-phenylalanine (l-dopa) is gaining interest as a drug of choice for parkinson's disease. aspergillus oryzae is commonly used for l-dopa production; however, a slower growth rate and relatively lower tyrosinase activity of mycelia have led to an increasing interest in exploiting alternative fungal cultures. in the present investigation, we report on the microbiological transformation of l-tyrosine to l-dopa accomplished by a newly isolated filamentous fung ...201021143944
selective induction, purification and characterization of a laccase isozyme from the basidiomycete trametes sp. ah28-2.the white-rot fungus trametes sp. ah28-2 can synthesize extracellular laccase by induction in cellobiose-based liquid culture medium. both yields and composition of laccase isozymes, produced by trametes sp. ah28-2, would be quite different with induction by different small-molecule aromatic compounds, o-toluidine, guaiacol and 3,5-dihydroxytoluene, which affected microbial growth and the synthesis of laccase isozymes differentially. higher concentrations of the three inducers could considerably ...200421148825
uniform culture in solid-state fermentation with fungi and its efficient enzyme production.solid-state fermentation (ssf) has attracted a lot of interest for carrying out high-level protein production in filamentous fungi. however, it has problems such as the fermentation heat generated during the culture in addition to the reduced mobility of substances. these conditions lead to a nonuniform state in the culture substrate and result in low reproducibility. we constructed a non-airflow box (nab) with a moisture permeable fluoropolymer membrane, thereby making it possible to control an ...201021163699
integrated analysis of the global transcriptional response to α-amylase over-production by aspergillus oryzae.the filamentous fungus aspergillus oryzae is an important microbial cell factory for industrial production of many enzymes, such as α-amylase. in order to optimize the industrial enzyme production process, there is a need to understand fundamental processes underlying enzyme production, here under how enzyme production links to metabolism through global regulation. through a genome-scale metabolic network for integrated analysis of transcriptome data and flux calculation, we identified key playe ...201021165907
molecular evolution and characterization of fungal indoleamine 2,3-dioxygenases.indoleamine 2,3-dioxygenase (ido) and tryptophan 2,3-dioxygenase (tdo) are tryptophan-degrading enzymes. mammalian ido expression is induced by cytokines and has antimicrobial and immunomodulatory effects. a major role of mammalian tdo is to supply nicotinamide adenine dinucleotide (nad(+)). in fungi, the ido homologue is thought to be expressed constitutively and supply nad(+), as tdo is absent from their genomes. here, we reveal the distribution of ido genes among fungal species and characteri ...201021170645
macroautophagy-mediated degradation of whole nuclei in the filamentous fungus aspergillus oryzae.filamentous fungi consist of continuum of multinucleate cells called hyphae, and proliferate by means of hyphal tip growth. accordingly, research interest has been focusing on hyphal tip cells, but little is known about basal cells in colony interior that do not directly contribute to proliferation. here, we show that autophagy mediates degradation of basal cell components in the filamentous fungus aspergillus oryzae. in basal cells, enhanced green fluorescent protein (egfp)-labeled peroxisomes, ...201021187926
regioselective deglycosylation of onion quercetin glucosides by saccharomyces cerevisiae.bioconversion of quercetin glucosides using four generally recognized as safe (gras) organisms (aspergillus oryzae, bacillus subtilis, lactobacillus plantarum, and saccharomyces cerevisiae) was evaluated by measuring changes in the levels of quercetin compounds of onion. of the four organisms, s. cerevisiae increased the content of quercetin-3-o-β-d-glucoside (iii; isoquercitrin) and quercetin (iv), whereas decreasing quercetin-3,4'-o-β-d-glucoside (i) and quercetin-4'-o-β-d-glucoside (ii). also ...201021188617
loss of aspergillus oryzae amyr function indirectly affects hemicellulolytic and cellulolytic enzyme production.aspergillus oryzae ab390, a derivative of a. oryzae or101, was found to be suitable for soy sauce production, yielding a product light brown in color. compared to the parent strain, hemicellulase and cellulase activities in the mutant were higher; however, its amylase activity was found to be much lower. to determine the cause of these differences, the enzymatic profile change, as a function of the carbon source in submerged cultures, was examined. amylase activity in ab390 was hardly detectable ...201021193346
effects of moisture enhancement, enzyme treatment, and blade tenderization on the processing characteristics and tenderness of beef semimembranosus steaks.the individual and combined effects of moisture enhancement with a salt/phosphate solution (me), blade tenderization (bt), and enzyme injection with proteinases derived from aspergillus oryzae or bacillus subtilis on cooking properties, warner-bratzler shear force (wbsf), and sensory characteristics of beef semimembranosus were investigated. me significantly (p < 0.01) reduced wbsf and increased (p < 0.05) sensory scores for juiciness and tenderness. bt increased (p < 0.05) initial and overall t ...201021194850
analysis of autophagy in aspergillus oryzae by disruption of aoatg13, aoatg4, and aoatg15 genes.autophagy is a degradation system in which cellular components are digested via vacuoles/lysosomes, and involved in differentiation in addition to helping cells to survive starvation. the autophagic process is composed of several steps: induction of autophagy, formation of autophagosomes, transportation to vacuoles, and degradation of autophagic bodies. to further understand autophagy in the filamentous fungus aspergillus oryzae, we first constructed a. oryzae mutants defective for the aoatg13, ...201121204928
post-translational his-cys cross-linkage formation in tyrosinase induced by copper(ii)-peroxo species.autocatalytic formation of his-cys cross-linkage in the enzyme active site of tyrosinase from aspergillus oryzae has been demonstrated to proceed by the treatment of apoenzyme with cu(ii) under aerobic conditions, where a (μ-η(2):η(2)-peroxo)dicopper(ii) species has been suggested to be involved as a key reactive intermediate.201121218798
characterization of an aspergillus oryzae cysteinyl dipeptidase expressed in escherichia coli.cysteinyl dipeptidase from aspergillus oryzae (cdpa) was produced in escherichia coli and purified. the enzyme showed activity specific toward cysteine-containing dipeptides, but its substrate specificity was distinct from those of other cysteinyl dipeptidases of the m20 family. it was optimally active at ph 7-8 and stable at ph 6-9 and at up to 40 °c.201121228467
five carboxin-resistant mutants exhibited various responses to carboxin and related fungicides.five carboxin-resistant mutants from aspergillus oryzae were characterized by the sensitivities of their mycelial growth and succinate dehydrogenase (sdh) activity to carboxin and three related fungicides. despite a significant resistance to carboxin, exhibited by all the mutants, their patterns of sensitivity to the other fungicides was distinct. this provides clues to the molecular interaction between sdh and these fungicides.201121228470
heterologous expression of aspergillus oryzae xylose reductase and xylitol dehydrogenase genes facilitated xylose utilization in the yeast saccharomyces cerevisiae.the cdna encoding a putative xylose reductase (xyra) from aspergillus oryzae was cloned and coexpressed in the yeast saccharomyces cerevisiae with a. oryzae xylitol dehydrogenase cdna (xdha). xyra exhibited nadph-dependent xylose reductase activity. the s. cerevisiae strain, overexpressing the xyra, xdha, endogenous xks1, and tal1 genes, grew on xylose as sole carbon source, and produced ethanol.201121228482
construction of a thiamine pyrophosphate high-producing strain of aspergillus oryzae by overexpression of three genes involved in thiamine biosynthesis.we have found a gene (thip) encoding thiamine pyrophosphokinase (tpk) in the aspergillus oryzae genome. no riboswitch-like region was found in the upstream region of thip, although it was repressed probably by thiamine pyrophosphate (tpp) as well as thia and nmta, which are strictly regulated by tpp-riboswitch sequence. to improve the productivity of tpp in a. oryzae, we constructed the strain in which thia, nmta and thip were overexpressed simultaneously. the resulting strain accumulated intrac ...201121247799
expression, purification and characterization of recombinant α-glucosidase in pichia pastoris.an expression plasmid containing the agda gene encoding aspergillus oryzae zl-1 α-glucosidase was constructed and expressed in pichia pastoris x-33. the molar mass of the purified protein was estimated by sds-page. hplc analysis showed that the purified enzyme has a transglucosylating activity with maltose as substrate. the main component of the enzyme products was panose, while amounts of isomaltose and isomaltotriose were very low or absent. ph 5.2 and temperature of 37 °c were optimum for enz ...201021253902
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