Changes in carbohydrase activities during vegetative growth and development of fruit-bodies of Hypsizygus marmoreus grown in sawdust-based culture

Carbohydraseproduction of Hypsizygus marmoreus cultured on sawdust rice bran medium by bottle cultivation was investigated to elucidate the carbohydrase utilized as the growth substrate for the fruit-body formation of this fungus. Among the extracellular enzymes assayed, xylanase showed the highest activity. This activity greatly increased during the end of vegetative mycelial growth and fruit-body formation. Among the cellulases. CM-cellulase showed higher activity than avicelase. The activities of -1,3-glucanase, amylase, and chitinase were low. Among the intracellular enzymes, both xylanase and amylase showed higher activity levels than the other carbohydrases. In contrast, -1,3-glucanase, avicelase, and chitinase activities in mycelia were considerably lower. These results suggest that xylanase, CM-cellulase, and amylase play an important role in mycelial maturation and fruit-body growth of H. marmoreus.     

An Improved Synthesis of 7, 8-Epoxy-1,3,11-cembratriene- 15R(α), 16-diol,a Cembranoid of Marine Origin with a Potent Cancer Chemopreventive Activity

An effective method for the synthesis of 7,8-epoxy-1,3,11-cembratriene- 15R(α),16-diol and its in vitro Epstein-Barr Virus Early Antigen (EBV-EA) Activation Chemopreventive Assay are reported. This semisynthetic product is a new cembranoid with a potent tumor inhibitory activity that is expected to be a lead compound for a new class of chemopreventive agents of marine origin.     

Optimum culture conditions of <Emphasis Type="Italic">Rhodomonas</Emphasis> sp. Hf-1 strain as a live food for aquatic animals

Suitable culture conditions for Rhodomonas sp. Hf-1 strain were examined for high productivity. Hf-1 strain was grown in an incubator for 7 days. The factorial experimental design investigated the following 19 variables: temperature (12, 16, 20, 24 and 28 °C), salinity (7, 14, 21, 28 and 35 psu), light intensity (20, 35, 50, 65 and 80 μmol m^?2 s^?1), light color (white, red, green and blue lights), and 3 factorial designs of photoperiod (24L:0D, 16L:8D and 12L:12D light:dark cycle). The cell density and specific growth rates (SGR) were analyzed. The best cell growth was observed under the following culture conditions: temperature of 24 °C, salinity of 21 psu, light intensity of 80 μmol m^?2 s^?1, and white light. In the photoperiod test, the highest cell density of 4.7?×?10⁶ cells ml^?1 was obtained at 24L:0D light:dark cycle, and the SGR was 0.57 μ day^?1 at this time. We found that the Hf-1 strain was able to be cultured in extremely wide culture conditions. These results are expected to serve as a baseline study for culturing the Hf-1 strain in the laboratory and for its use in aquaculture.     

Identification of a 4-Deoxy-l-erythro-5-hexoseulose Uronic Acid Reductase,FlRed, in an Alginolytic Bacterium Flavobacterium sp. Strain UMI-01

In alginate-assimilating bacteria, alginate is depolymerized to unsaturated monosaccharide by the actions of endolytic and exolytic alginate lyases (EC 4.2.2.3 and EC 4.2.2.11). The monosaccharide is non-enzymatically converted to 4-deoxy-l-erythro-5-hexoseulose uronic acid (DEH), then reduced to 2-keto-3-deoxy-d-gluconate (KDG) by a specific reductase, and metabolized through the Entner–Doudoroff pathway. Recently, the NADPH-dependent reductase A1-R that belongs to short-chain dehydrogenases/reductases (SDR) superfamily was identified as the DEH-reductase in Sphingomonas sp. A1. We have subsequently noticed that an SDR-like enzyme gene, flred, occurred in the genome of an alginolytic bacterium Flavobacterium sp. strain UMI-01. In the present study, we report on the deduced amino-acid sequence of flred and DEH-reducing activity of recombinant FlRed. The deduced amino-acid sequence of flred comprised 254 residues and showed 34% amino-acid identities to that of A1-R from Sphingomonas sp. A1 and 80%–88% to those of SDR-like enzymes from several alginolytic bacteria. Common sequence motifs of SDR-superfamily enzymes, e.g., the catalytic tetrad Asn-Lys-Tyr-Ser and the cofactor-binding sequence Thr-Gly-x-x-x-Gly-x-Gly in Rossmann fold, were completely conserved in FlRed. On the other hand, an Arg residue that determined the NADPH-specificity of Sphingomonas A1-R was replaced by Glu in FlRed. Thus, we investigated cofactor-preference of FlRed using a recombinant enzyme. As a result, the recombinant FlRed (recFlRed) was found to show high specificity to NADH. recFlRed exhibited practically no activity toward variety of aldehyde, ketone, keto ester, keto acid and aldose substrates except for DEH. On the basis of these results, we conclude that FlRed is the NADH-dependent DEH-specific SDR of Flavobacterium sp. strain UMI-01.     

Culture method and growth characteristics of marine benthic dinoflagellate Ostreopsis spp. isolated from Japanese coastal waters

Blooms of toxic dinoflagellates of the genus Ostreopsis, which is known as a producer of palytoxin (PTX) analogs, may pose a threat to human health in tropical, subtropical, and temperate regions around the world. In the present study, we established a suitable culture method for Ostreopsis spp. isolated from Japanese coastal waters and characterized their growth potential using the method to discuss their bloom dynamics. Each clonal strain of Ostreopsis cf. ovata, Ostreopsis sp.?1, Ostreopsis sp.?5, and Ostreopsis sp.?6 was incubated in 25?×?150?mm test tubes with a flat bottom containing various kinds of medium. Since Ostreopsis spp. strains grew well in IMK and/or f/2 media, we selected these media for cultivation of all the Ostreopsis spp. isolates. Growth rates of O. cf. ovata (0.834?divisions/day), Ostreopsis sp.?1 (0.619?divisions/day), and Ostreopsis sp.?6 (1.04?divisions/day) that produce PTX analogs significantly differed (p?<?0.05) respectively and are clearly higher than those of other reported epiphytic dinoflagellate Gambierdiscus toxicus, Prorocentrum lima, and Coolia monotis cultures, which suggest that these species have ecological advantages to predominate through the algal succession in Japanese coastal waters, resulting in a potential risk to human health in this region.     

Microbial reduction mechanism of ferric iron in paddy soils (Part I)

In the previous paper¹⁾ the authors proved that the ferric iron reduction in submerged paddy soils is entirely or mostly effected by the activities of microorganisms in the soil. Prior to the study on the ferric iron reducing microorganisms isolated from the soil, the present paper reports the research for the microbial mechanism of iron reduction. working with the soil itself.