Kaori Tomita-Yokotani

Discovery of the curcumin metabolic pathway involving a unique enzyme in an intestinal microorganism

Polyphenol curcumin, a yellow pigment, derived from the rhizomes of a plant (Curcuma longa Linn) is a natural antioxidant exhibiting a variety of pharmacological activities and therapeutic properties. It has long been used as a traditional medicine and as a preservative and coloring agent in foods. Here, curcumin-converting microorganisms were isolated from human feces, the one exhibiting the highest activity being identified as Escherichia coli. We are thus unique in discovering that E. coli was able to act on curcumin. The curcumin-converting enzyme was purified from E. coli and characterized. The native enzyme had a molecular mass of about 82 kDa and consisted of two identical subunits. The enzyme has a narrow substrate spectrum, preferentially acting on curcumin. The microbial metabolism of curcumin by the purified enzyme was found to comprise a two-step reduction, curcumin being converted NADPH-dependently into an intermediate product, dihydrocurcumin, and then the end product, tetrahydrocurcumin. We named this enzyme “NADPH-dependent curcumin/dihydrocurcumin reductase” (CurA). The gene (curA) encoding this enzyme was also identified. A homology search with the BLAST program revealed that a unique enzyme involved in curcumin metabolism belongs to the medium-chain dehydrogenase/reductase superfamily.

A species-selective allelopathic substance from germinating sunflower (Helianthus annuus L.) seeds

From the exudate of germinating sunflower (Helianthus annuus L.) seeds was isolated a stereoisomer of diversifolide, 4, 15-dinor-3-hydroxy-1(5)-xanthene-12,8-olide (designated sundiversifolide) as determined by analysis of its IR, APCI-, ESI- and HR-MS and 13C and 1H NMR spectra. This substance inhibited shoot and root growth of cats-eyes by about 50% at a concentration of 30 ppm. It also showed species-selective activity on the shoot and root growth of tested plants. When cats-eyes seeds were incubated together with sunflower seeds, the cats-eyes growth was inhibited. Furthermore, it was detected from an extract of river sand when sunflower seeds were incubated on the sand. These results indicate that sundiversifolide has an allelopathic function in sunflower plants.

Discovery of Amide (Peptide) Bond Synthetic Activity in Acyl-CoA Synthetase

Acyl-CoA synthetase, which is one of the acid-thiol ligases (EC 6.2.1), plays key roles in metabolic and regulatory processes. This enzyme forms a carbon-sulfur bond in the presence of ATP and Mg2+, yielding acyl-CoA thioesters from the corresponding free acids and CoA. This enzyme belongs to the superfamily of adenylate-forming enzymes, whose three-dimensional structures are analogous to one another. We here discovered a new reaction while studying the short-chain acyl-CoA synthetase that we recently reported (Hashimoto, Y., Hosaka, H., Oinuma, K., Goda, M., Higashibata, H., and Kobayashi, M. (2005) J. Biol. Chem. 280, 8660–8667). When l-cysteine was used as a substrate instead of CoA, N-acyl-l-cysteine was surprisingly detected as a reaction product. This finding demonstrated that the enzyme formed a carbon-nitrogen bond (EC 6.3.1 acid-ammonia (or amide) ligase (amide synthase); EC 6.3.2 acid-amino acid ligase (peptide synthase)) comprising the amino group of the cysteine and the carboxyl group of the acid. N-Acyl-d-cysteine, N-acyl-dl-homocysteine, and N-acyl-l-cysteine methyl ester were also synthesized from the corresponding cysteine analog substrates by the enzyme. Furthermore, this unexpected enzyme activity was also observed for acetyl-CoA synthetase and firefly luciferase, indicating the generality of the new reaction in the superfamily of adenylate-forming enzymes.

An overview of challenges in modeling heat and mass transfer for living on Mars.

Abstract:  Engineering a life‐support system for living on Mars requires the modeling of heat and mass transfer. This report describes the analysis of heat and mass transfer phenomena in a greenhouse dome, which is being designed as a pressurized life‐support system for agricultural production on Mars. In this Martian greenhouse, solar energy will be converted into chemical energy in plant biomass. Agricultural products will be harvested for food and plant cultivation, and waste materials will be processed in a composting microbial ecosystem. Transpired water from plants will be condensed and recycled. In our thermal design and analysis for the Martian greenhouse, we addressed the question of whether temperature and pressure would be maintained in the appropriate range for humans as well as plants. Energy flow and material circulation should be controlled to provide an artificial ecological system on Mars. In our analysis, we assumed that the greenhouse would be maintained at a subatmospheric pressure under 1/3‐G gravitational force with 1/2 solar light intensity on Earth. Convection of atmospheric gases will be induced inside the greenhouse, primarily by heating from sunlight. Microclimate (thermal and gas species structure) could be generated locally around plant bodies, which would affect gas transport. Potential effects of those environmental factors are discussed on the phenomena including plant growth and plant physiology and focusing on transport processes. Fire safety is a crucial issue and we evaluate its impact on the total gas pressure in the greenhouse dome.

Plant growth-promoting oligosaccharides produced from tomato waste

Tomato juice waste was hydrolyzed with acid. Tomato juice waste (500 g; wet weight) was heated with 0.5 N HCl (2.5 l) at 70 degrees C for 4 h. After neutralization, the growth-promoting extracts (300 g; dry weight) in the plants were produced from the tomato waste. The acid extract significantly promoted the growth of cockscomb (Celosia argentea L.) and tomato (Lycopersicon esculentum L.) seedlings. We have recognized potent plant growth-promoting substances in the acid extract from tomato waste. The most effective components in the active fraction were almost all oligogalacturonic acids (DP 6-12). This paper is the first report that plant growth-promoting oligosaccharides can be directly produced from tomato juice waste. It is possible that the substances from the tomato waste can become useful plant growth regulators in the agriculture field in the future.

Isolation and purification of an axenic diazotrophic drought-tolerant cyanobacterium, Nostoc commune, from natural cyanobacterial crusts and its utilization for field research on soils polluted with radioisotopes.

Nitrogen fixation and drought tolerance confer the ability to grow on dry land, and some terrestrial cyanobacteria exhibit these properties. These cyanobacteria were isolated in an axenic form from Nostoc commune clusters and other sources by modifying the method used to isolate the nitrogen-fixing and drought-tolerant cyanobacterium Nostoc sp. HK-01. Of these cyanobacteria, N. commune, which is difficult to isolate and purify, uses polysaccharides to maintain water, nitrogen fertilizers for nitrogen fixation, and can live in extreme environments because of desiccation tolerance. In this study, we examined the use of N. commune as biosoil for space agriculture and possible absorption of radioisotopes ((134)Cs, (137)Cs). This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.

Flavonoids in the leaves of Oxalis corniculata and sequestration of the flavonoids in the wing scales of the pale grass blue butterfly, Pseudozizeeria maha

Three C-glycosylflavones in the leaves of Oxalis corniculata, the host plant of the lycaenid butterfly pale grass blue (Pseudozizeeria maha), were identified as 6-C-glucosylluteolin (isoorientin), 6-C-glucosylapigenin (isovitexin) and isovitexin 7-methyl ether (swertisin). Comparative spectral and HPLC analyses between the leaf extract of the host plants and the wings of P. maha showed selective uptake of the host-plant flavonoid isovitexin to the wings of the butterfly.

Preparation and Isolation of Oligogalacturonic Acids and Their Biological Effects in Cockscomb (Celosia argentea L.) Seedlings

AbstractA mixture of oligogalacturonic acids, the partial degradation substances of polygalacturonic acid, promoted shoot growth in cockscomb (Celosia argentea L.) seedlings, which generally had a high sensitivity for growth-promoting substances. The effect of the mixture of oligogalacturonic acids on shoot growth of cockscomb was higher than that of the polygalacturonic acid at concentrations above 30 ppm. These oligomers were loaded onto an anion exchange column, DEAE Sephadex A-25, and separated into individual oligomer sizes using the NH4HCO3 eluent system. This separation method has the advantage of using NH4HCO3 as the eluent solution; NH4HCO3 in the sample solution is effectively removed by lyophilization. Each of the isolated oligogalacturonic acids gave a single band on a fluorophore-assisted carbohydrate electrophoresis (FACE), and they showed the m/z value which corresponded to their molecular ion peaks [M-H]- on a fast atom bombardment mass spectrometry (FAB-MS) analysis. These results showed that the successive chromatography method used in this study is well suited for the preparation of oligogalacturonic acid for the plant growth test. Furthermore, we showed that the effective degree of polymerization (DP) of oligogalacturonic acid was around 8 on shoot growth of cockscomb seedlings, and the effects of both smaller and larger oligogalacturonic acids were slightly lower than that of octa-galacturonide. Octa-galacturonide promoted shoot growth of cockscomb at concentrations above 10 mM, and showed a 66% promotion at the most effective concentration of 300 mM. Root growth was slightly inhibited at concentrations above 300 mM. These results suggest that DP around 8 of oligogalacturonic acids has the function to control shoot growth in cockscomb as a growth-promoting substance.

Improvement of Bioactive Potential of Canavalia Beans of Coastal Sand Dunes by Solid-Substrate Fermentation Using Rhizopus oligosporus

Cooked beans of two wild legumes (Canavalia cathartica and C. maritima) were subjected to solid-substrate fermentation by Rhizopus oligosporus and bioactive potential has been compared with raw and cooked beans. Flavonoids and canavanine in raw beans were significantly decreased on cooking, while they were significantly enhanced in fermented beans. Phytic acid content was signficantlty decreased from raw vs. cooked vs. fermented beans. Raw, cooked and fermented beans were devoid of trypsin inhibitors. Hemagglutinin activity of raw beans was completely eliminated on cooking, while SSF increased its activity about 50% of raw beans against A+ve and or B+ve blood groups. Solid-substrate fermentation of cooked beans with R. oligosporus resulted in better nutraceutical potential compared to raw and mearly cooked beans qualifying as important nutraceutical source to develop indigenous value-added products.

Bioactive potential of the wild mushroom Astraeus hygrometricus in South-west India

ABSTRACT The wild mushroom Astraeus hygrometricus is a traditional delicacy during the monsoon season in South-western India. Bioactive principles and antioxidant potential of uncooked and pressure-cooked tender mushroom have been evaluated. Seven bioactive principles of mushroom (tannins, flavonoids, vitamin C, phytic acid, lycopene, β-carotene and trypsin inhibition) were significantly higher, while total phenolics content was significantly lower in uncooked than in cooked samples. Mushroom was devoid of L-DOPA, whereas only uncooked samples showed haemagglutinin activity against A+ blood group. The principal component analysis of uncooked mushroom showed only two bioactive principles clustered with two antioxidant properties, while in cooked mushroom five bioactive principles clustered with three antioxidant properties depicting the nutraceutical potential of cooked mushroom. Future studies should focus on appropriate thermal treatment, which retain maximum bioactive and antioxidant potential to combat health- and lifestyle-related ailments. The A. hygrometricus is ectomycorrhizal, conservation of its host tree species is utmost importance in improvement and expansion of its yield to sustain food security and economic gains of local tribals.