Production and properties of a laccase isozyme (FvLcc3) from the edible mushroom Flammulina velutipes, which is undetectable under the culture condition for fruiting body formation.

Extracellular laccase isozyme (FvLcc3) from the edible mushroom Flammulina velutipes was found to be undetectable under the culture condition for fruiting body formation. FvLcc3 was purified and determined to be an approximately 53-kDa monomeric protein. FvLcc3 showed the highest catalytic efficiency (kcat/Km) toward 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) followed by 2,6-dimethoxyphenol and guaiacol and did not oxidize 3,4-dihydroxy-l-phenylalanine and l-tyrosine.

Relationship between fruiting body development and extracellular laccase production in the edible mushroom .

The biochemical mechanism underlying the development of fruiting bodies in , an edible mushroom, was investigated using the YBLB colorimetric assay to distinguish between the normal strain (FVN-1) and the degenerate strain (FVD-1). In this assay, the color of the YBLB medium (blue-green) inoculated with FVN-1 exhibiting normal fruiting body development changed to yellow, while the color of the medium inoculated with FVD-1 changed to blue. In this study, we found that this color difference originated from extracellular laccase produced by FVN-1.

Biochemical synthesis of the medicinal sugar l-gulose using fungal alditol oxidase.

Some rare sugars can be potently medicinal, such as l-gulose. In this study, we present a novel alditol oxidase (fAldOx) from the soil fungus Penicillium sp. KU-1, and its application for the effective production of l-gulose.

Efficient production of the rare sugar l-gulose using a wheat-bran culture extract of Penicillium sp. KU-1.

We found that l-gulose, a rare sugar, was produced from d-sorbitol efficiently, using a wheat-bran culture extract of the fungus Penicillium sp. KU-1 isolated from soil. The culture extract showed enzyme activity for the oxidation of d-sorbitol to produce l-gulose; a high production yield of approximately 94% was achieved.

Heterologous expression of two minor laccase isozyme cDNAs from the edible mushroom Grifola frondosa.

Two cDNAs encoding the minor laccase isozymes (Lac2 and Lac3) of Grifola frondosa were cloned, characterized, and expressed in Pichia pastoris. The recombinant Lac2 (rLac2) was stable at pH 6.0, whereas the recombinant Lac3 (rLac3) was stable in a broad pH range (pH 4.

Occurrence of the (2R,3S)-Isomer of 2-Amino-3,4-dihydroxybutanoic Acid in the Mushroom Hypsizygus marmoreus.

Here, we report the occurrence of the (2R,3S)-isomer of 2-amino-3,4-dihydroxybutanoic acid (d-ADHB) in the fruiting body of an edible mushroom, Hypsizygus marmoreus. This is an unusual example of the accumulation of a d-amino acid whose enantiomer is not a proteinogenic amino acid. We show that d-ADHB occurs specifically in the mushroom H.

Novel process for producing 6-deoxy monosaccharides from l-fucose by coupling and sequential enzymatic method.

We biosynthesized 6-deoxy-L-talose, 6-deoxy-L-sorbose, 6-deoxy-L-gulose, and 6-deoxy-L-idose, which rarely exist in nature, from L-fucose by coupling and sequential enzymatic reactions. The first product, 6-deoxy-L-talose, was directly produced from L-fucose by the coupling reactions of immobilized D-arabinose isomerase and immobilized L-rhamnose isomerase. In one-pot reactions, the equilibrium ratio of L-fucose, L-fuculose, and 6-deoxy-L-talose was 80:9:11.

Skipping Breakfast is Correlated with Obesity.

Objective: Despite the fact that the total energy intake of Japanese people has decreased, the percentage of obese people has increased. This suggests that the timing of meals is related to obesity. The purpose of the study was to investigate the relationship between the timing of meals and obesity, based on analyses of physical measurements, serum biochemical markers, nutrient intake, and lifestyle factors in the context of Chrononutrition.

Enzymatic production of three 6-deoxy-aldohexoses from L-rhamnose.

6-Deoxy-L-glucose, 6-deoxy-L-altrose, and 6-deoxy-L-allose were produced from L-rhamnose with an immobilized enzyme that was partially purified (IE) and an immobilized Escherichia coli recombinant treated with toluene (TT). 6-Deoxy-L-psicose was produced from L-rhamnose by a combination of L-rhamnose isomerase (TT-PsLRhI) and D-tagatose 3-epimerase (TT-PcDTE). The purified 6-deoxy-L-psicose was isomerized to 6-deoxy-L-altrose and 6-deoxy-L-allose with L-arabinose isomerase (TT-EaLAI) and L-ribose isomerase (TT-AcLRI), respectively, and then was epimerized to L-rhamnulose with immobilized D-tagatose 3-epimerase (IE-PcDTE).

Purification and characterization of the extracellular laccase produced by Trametes polyzona WR710-1 under solid-state fermentation.

Laccase from Trametes polyzona WR710-1 was produced under solid-state fermentation using the peel from the Tangerine orange (Citrus reticulata Blanco) as substrate, and purified to homogeneity. This laccase was found to be a monomeric protein with a molecular mass of about 71 kDa estimated by SDS-PAGE. The optimum pH was 2.

Daily rice intake strongly influences the incidence of metabolic syndrome in Japanese men aged 40-59 years.

Objectives: The first objective of this study was to classify men aged 40-74 yrs with metabolic syndrome (MetS) according to daily rice intake, and the second was to investigate physical measurements, physiological examinations, blood biochemical assays, intake of food other than rice and lifestyle and environmental factors in the study group.

Methods: We analyzed data from 6095 men aged 40-74 yrs who had undergone full medical examinations. The men were classified into 3 age groups: (1) 40-49 yrs, (2) 50-59 yrs, and (3) 60-74 yrs.

Comparison of regional differences in health indicators and standard mortality ratio for stroke in subjects in ehime prefecture.

Objective: The purpose of this study was to investigate regional differences in the standard mortality ratio (SMR) and risk factors (including dietary habits) for stroke across the three regions of Ehime Prefecture - Toyo (east), Chuyo (central), and Nanyo (south).

Participants And Methods: We obtained medical records derived from 956,979 medical examinations carried out at JA Ehime Kouseiren Medical Examination Centers between April 1994 and March 2006. We analyzed data from 132,090 subjects (Toyo - 47,654, Chuyo - 38,435, Nanyo - 46,001) who underwent their first medical examination during this period.

Biodegradation of bisphenol A and decolorization of synthetic dyes by laccase from white-rot fungus, Trametes polyzona.

Purified laccase from Trametes polyzona WR710-1 was used as biocatalyst for bisphenol A biodegradation and decolorization of synthetic dyes. Degradation of bisphenol A by laccase with or without redox mediator, 1-hydroxybenzotriazole (HBT) was studied. The quantitative analysis by HPLC showed that bisphenol A rapidly oxidized by laccase with HBT.

A cross-sectional survey of factors influencing bone mass in junior high school students.

Objective: The purpose of this study is to investigate factors influencing the osteo-sono assessment index (OSI) in junior high school students (boys, girls who had reached menarche, and girls who had not).

Methods: A total of 9,743 students (4,974 boys and 4,769 girls) in Ehime Prefecture participated in this study. We measured body mass index (BMI) and calcaneal bone mass using OSI.

Decolorization of synthetic dyes and biodegradation of bisphenol a by laccase from the edible mushroom, Grifola frondosa.

A major laccase isozyme from Grifola frondosa (Lac 1) was found to be effective for decolorizing of synthetic dyes and degrading of bisphenol A. The oxidative capability of Lac 1 toward synthetic dyes and bisphenol A was enhanced in the presence of the redox mediator, 1-hydroxybenzotriazole. The major product from the degradation of bisphenol A by Lac 1 was determined to be 4-isopropenylphenol.

CoPK32 is a novel stress-responsive protein kinase in the mushroom Coprinopsis cinerea.

Background: In a previous study, we conducted an expression cloning screen of a cDNA library prepared from Coprinopsis cinerea mycelia using Multi-PK antibodies and detected a wide variety of Ser/Thr protein kinases. One of the isolated clones, CMZ032, was found to encode a putative Ser/Thr protein kinase designated CoPK32. In the present study, we investigated the biochemical properties and physiological significance of CoPK32.

Extracellular laccase produced by an edible basidiomycetous mushroom, Grifola frondosa: purification and characterization.

A major laccase isozyme (Lac 1) was isolated from the culture fluid of an edible basidiomycetous mushroom, Grifola frondosa. Lac 1 was revealed to be a monomeric protein with a molecular mass of 71 kDa. The N-terminal amino acid sequence of Lac 1 was highly similar to those of laccases of some other white-rot basidiomycetes.

Catalytic properties and crystal structure of quinoprotein aldose sugar dehydrogenase from hyperthermophilic archaeon Pyrobaculum aerophilum.

We identified a gene encoding a soluble quinoprotein glucose dehydrogenase homologue in the hyperthermophilic archaeon Pyrobaculum aerophilum. The gene was overexpressed in Escherichia coli, after which its product was purified and characterized. The enzyme was extremely thermostable, and the activity of the pyrroloquinoline quinone (PQQ)-bound holoenzyme was not lost after incubation at 100 degrees C for 10 min.

Novel Ca2+/calmodulin-dependent protein kinase expressed in actively growing mycelia of the basidiomycetous mushroom Coprinus cinereus.

We isolated cDNA clones for novel protein kinases by expression screening of a cDNA library from the basidiomycetous mushroom Coprinus cinereus. One of the isolated clones was found to encode a calmodulin (CaM)-binding protein consisting of 488 amino acid residues with a predicted molecular weight of 53,906, which we designated CoPK12. The amino acid sequence of the catalytic domain of CoPK12 showed 46% identity with those of rat Ca2+/CaM-dependent protein kinase (CaMK) I and CaMKIV.

Involvement of Ca(2+)/calmodulin-dependent protein kinases in mycelial growth of the basidiomycetous mushroom, Coprinus cinereus.

Although multifunctional Ca(2+)/calmodulin-dependent protein kinases (CaM-kinases) are widely distributed in animal cells, the occurrence of CaM-kinases in the basidiomycetous mushroom has not previously been documented. When the extracts from various developmental stages from mycelia to the mature fruiting body of Coprinus cinereus were analyzed by Western blotting using Multi-PK antibodies, which had been generated to detect a wide variety of protein serine/threonine kinases (Ser/Thr kinases), a variety of stage-specific Ser/Thr kinases was detected. Calmodulin (CaM) overlay assay using digoxigenin-labeled CaM detected protein bands of 65 kDa, 58 kDa, 46 kDa, 42 kDa, and 38 kDa only in the presence of CaCl(2), suggesting that these bands were CaM-binding proteins.