Osamu Takimura

Bioethanol production performance of five recombinant strains of laboratory and industrial xylose-fermenting Saccharomyces cerevisiae.

In this study, five recombinant Saccharomyces cerevisiae strains were compared for their xylose-fermenting ability. The most efficient xylose-to-ethanol fermentation was found by using the industrial strain MA-R4, in which the genes for xylose reductase and xylitol dehydrogenase from Pichia stipitis along with an endogenous xylulokinase gene were expressed by chromosomal integration of the flocculent yeast strain IR-2. The MA-R4 strain rapidly converted xylose to ethanol with a low xylitol yield. Furthermore, the MA-R4 strain had the highest ethanol production when fermenting not only a mixture of glucose and xylose, but also mixed sugars in the detoxified hydrolysate of wood chips. These results collectively suggest that MA-R4 may be a suitable recombinant strain for further study into large-scale ethanol production from mixed sugars present in lignocellulosic hydrolysates.

Production of free and organic iodine by Roseovarius spp.

Two strains of iodine-producing bacteria were isolated from marine samples. 16S rRNA gene sequences indicated the strains were most closely related to Roseovarius tolerans, and phylogenetic analysis indicated both belong to the same genus. 5 mM iodide inhibited the growth of strain 2S5-2 almost completely, and of strain S6V slightly. Both strains produced free iodine and organic iodine from iodide. CH2I2, CHI3 and CH2ClI were the main organic iodines produced by strain 2S5-2, and CHI3 and CH2I2 by strain S6V. Experiments using cells and spent media suggested that the organic iodines were produced from the compounds released or contained in the media and cells were necessary for the considerable production of CH2I2 and CH2ClI, though CHI3 was produced by spent media with H2O2 or free iodine.

Degradation of triphenyltin by a fluorescent pseudomonad.

ABSTRACT Triphenyltin (TPT)-degrading bacteria were screened by a simple technique using a post-column high-performance liquid chromatography using 3,3′,4′,7-tetrahydroxyflavone as a post-column reagent for determination of TPT and its metabolite, diphenyltin (DPT). An isolated strain, strain CNR15, was identified as Pseudomonas chlororaphis on the basis of its morphological and biochemical features. The incubation of strain CNR15 in a medium containing glycerol, succinate, and 130 μM TPT resulted in the rapid degradation of TPT and the accumulation of approximately 40 μM DPT as the only metabolite after 48 h. The culture supernatants of strain CNR15, grown with or without TPT, exhibited a TPT degradation activity, whereas the resting cells were not capable of degrading TPT. TPT was stoichiometrically degraded to DPT by the solid-phase extract of the culture supernatant, and benzene was detected as another degradation product. We found that the TPT degradation was catalyzed by low-molecular-mass substances (approximately 1,000 Da) in the extract, termed the TPT-degrading factor. The other fluorescent pseudomonads,P. chlororaphis ATCC 9446, Pseudomonas fluorescens ATCC 13525, and Pseudomonas aeruginosaATCC 15692, also showed TPT degradation activity similar to strain CNR15 in the solid-phase extracts of their culture supernatants. These results suggest that the extracellular low-molecular-mass substance that is universally produced by the fluorescent pseudomonad could function as a potent catalyst to cometabolite TPT in the environment.

Effect of Glutathione on Arsenic Accumulation by Dunaliella salina

The behavior of marine algae (Dunaliella salina, Chattonella antiqua, Heteresigma akashiwo, Skeletonema costatum, chaetoceros debile and Thalassiosira weissflogii) against arsenate, arsenite and DMA in a medium and the effects of glutathione that influenced the redox condition on arsenic accumulation of D. salina were studied. It was found that the order of growth inhibition of marine algae by arsenic species was As(III) > As(V) > DMA. The order of arsenic accumulation by D. salina was As(V) ≥ As(III) > DMA at a concentration of 100 mgAsdm -3 . A small part of the arsenic accumulated by D. salina was methylated in vivo. DMA was the major methylated arsenic compound. Methylated arsenic compounds were not present in the medium. Glutathione (GSH) treatment increased arsenic accumulation by D. salina at a concentration of 10-100 mg GSHdm -3 . Buthionine sulfoxisamine (a potent and specific inhibitor of γ-glutamylcysteine synthetase) strongly suppressed the effect of GSH on arsenic accumulation. These findings suggest that the intracellular glutathione concentration may be important for arsenic accumulation.

Tin-Carbon Cleavage of Organotin Compounds by Pyoverdine from Pseudomonas chlororaphis

ABSTRACT The triphenyltin (TPT)-degrading bacterium Pseudomonas chlororaphis CNR15 produces extracellular yellow substances to degrade TPT. Three substances (F-I, F-IIa, and F-IIb) were purified, and their structural and catalytic properties were characterized. The primary structure of F-I was established using two-dimensional nuclear magnetic resonance techniques; the structure was identical to that of suc-pyoverdine from P. chlororaphis ATCC 9446, which is a peptide siderophore produced by fluorescent pseudomonads. Spectral and isoelectric-focusing analyses revealed that F-IIa and F-IIb were also pyoverdines, differing only in the acyl substituent attached to the chromophore part of F-I. Furthermore, we found that the fluorescent pseudomonads producing pyoverdines structurally different from F-I showed TPT degradation activity in the solid extracts of their culture supernatants. F-I and F-IIa degraded TPT to monophenyltin via diphenyltin (DPT) and degraded DPT and dibutyltin to monophenyltin and monobutyltin, respectively. The total amount of organotin metabolites produced by TPT degradation was nearly equivalent to that of the F-I added to the reaction mixture, whereas DPT degradation was not influenced by monophenyltin production. The TPT degradation activity of F-I was remarkably inhibited by the addition of metal ions chelated with pyoverdine. On the other hand, the activity of DPT was increased 13- and 8-fold by the addition of Cu2+ and Sn4+, respectively. These results suggest that metal-chelating ligands common to pyoverdines may play important roles in the Sn-C cleavage of organotin compounds in both the metal-free and metal-complexed states.

Utilization of Dimethyl Sulfide as a Sulfur Source with the Aid of Light by Marinobacterium sp. Strain DMS-S1

ABSTRACT Strain DMS-S1 isolated from seawater was able to utilize dimethyl sulfide (DMS) as a sulfur source only in the presence of light in a sulfur-lacking medium. Phylogenetic analysis based on 16S ribosomal DNA genes indicated that the strain was closely related toMarinobacterium georgiense. The strain produced dimethyl sulfoxide (DMSO), which was a main metabolite, and small amounts of formate and formaldehyde when grown on DMS as the sole sulfur source. The cells of the strain grown with succinate as a carbon source were able to use methyl mercaptan or methanesulfonate besides DMS but not DMSO or dimethyl sulfone as a sole sulfur source. DMS was transformed to DMSO primarily at wavelengths between 380 and 480 nm by heat-stable photosensitizers released by the strain. DMS was also degraded to formaldehyde in the presence of light by unidentified heat-stable factors released by the strain, and it appeared that strain DMS-S1 used the degradation products, which should be sulfite, sulfate, or methanesulfonate, as sulfur sources.

Uptake and Reduction of Arsenate by Dunaliella sp.

Uptake and reduction of arsenate [AS(V)] by Dunaliella sp. cells were determined to investigate the metabolic processes of arsenic in the alga. Cellular uptake of arsenic by Dunaliella sp. cells was markedly affected by the form of arsenic in the medium. The content of arsenic taken up by Dunaliella sp. cells increased rapidly with time on addition of As(V) to the medium. However, in the case of addition of arsenite [As(III)], the gradient of arsenic uptake by Dunaliella sp. cells was low, and arsenic content was small. In the water-soluble fraction of arsenic taken up by Dunaliella sp. cells with exposure to As(V), arsenic was in the forms of organic arsenic, As(V) and As(III). The content of As(V) in the water-soluble fraction increased with exposure time. The content of As(III) also increased with time, but remained constant after 5h of exposure. On the other hand, organic arsenic content was small and did not increase with time. It was found that Dunaliella sp. takes up As(V) and readily reduces it to As(III).

Vertical distribution of particulate manganese and iron in the Beppu Bay

Around the thermocline, difference in the depth of the maximum concentration between particulate manganese and iron was observed above the basin located at the south-western corner of the Beppu Bay, along with the remarkable difference in pH and Eh.

Pressure-Induced Alteration in Fatty Acid Composition of Barotolerant Deep-Sea Bacterium

Barotolerant bacterium was isolated from sediment sample which was obtained from the depth of 4033 m in the Izu-Ogasawara Trench. The physiological property, growth characteristics and fatty acid composition were examined. The strain was a psychrotrophic and barotolerant bacterium, and was identified as species in the genusAlteromonas. The fatty acids of the strain were from C12 to C18. As the growth pressure increased, the portion of unsaturated fatty acid in membrane fraction increased due to an increase in the portion of C17∶1 and C18∶1, while the relative portion of C16∶0 and C16∶1 decreased. On the other hand, as the growth temperature decreased, the portion of unsaturated fatty acid increased due to the increase in the portion of C16∶1 and C18∶1.

Vertical distribution of heavy metals and its seasonal variations in Beppu Bay

From July to November, the thermocline which has strong temperature gradient (0.7‡C m−1) is formed in the bottom water of Beppu Bay, and it prevents the downward mixing of surface water. This has caused the bottom water of the basin to become depleted in oxygen, and in November the bottom water below about 60 m depth becomes anoxic. Accordingly manganese and iron are reduced and more soluble under the anoxic condition, those concentrations are high relative to surface water, and the maximums are 1,240Μg l−1 and 80Μg l−1. Under the anoxic condition, the flux of dissolved manganese from the sediment is about 10Μg cm−2 day−1.