Isolation of dimethyl sulfone-degrading microorganisms and application to odorless degradation of dimethyl sulfoxide

With the objective of developing an odorless biodegradation process for dimethyl sulfoxide (DMSO), Hyphomicrobium sp. WU-OM3 was isolated. During the cultivation of strain WU-OM3 cells with 20 mM dimethyl sulfone (DMSO2) as the sole carbon source, DMSO2 was completely consumed within 48 h and sulfate ion accumulated in the culture broth. Methanesulfonate was also detected as an intermediate of DMSO2 degradation. By combining the DMSO-oxidizing microorganism and strain WU-OM3 cells, 0.64 mM (50 mg/l) DMSO was degraded to sulfate ion with 80% molar conversion ratio.     

Cloning and characterization of a β-N-acetylglucosaminidase (BmFDL) from silkworm Bombyx mori

In insects, β-N-acetylglucosaminidase (GlcNAcase) participates in critical physiological processes such as fertilization, metamorphosis, and glycoconjugate degradation. Insects produce glycoproteins carrying paucimannosidic-type N-glycans, the terminal GlcNAc residue of which is cleaved by a GlcNAc-linkage specific GlcNAcase, also known as the fused lobes (FDL) protein. To obtain information on the structure of GlcNAcases and insight into their contribution to physiological processes, we cloned Bombyx mori FDL (BmFDL) from silkworm larvae. The full-length cDNA (1.9 kb) encoded a protein of 633 amino acids with 42% amino acid sequence identity to Drosophila melanogaster FDL (DmFDL). Recombinant BmFDL cleaved only β-1,2-linked GlcNAc residues from the α-1,3 branch of biantennary N-glycan. This substrate specificity was similar to that of DmFDL. Microsomal FDL activity was inhibited by anti-BmFDL antibodies. Taken together, our results suggest that BmFDL is a N-glycan-processing GlcNAcase in B. mori.     

Simultaneous estimation of the thermophysical properties of three kinds of fruit juices based on the measured result by a transient heat flow probe method

The thermophysical properties of three kinds of fruit juices (grape juice, orange juice, and pineapple juice) were measured at various temperatures (10–50 °C) and concentrations (10–50%). A new method for the simultaneous determination of thermophysical properties using a modified version of current probe theory method was proposed. The temperature changes of the probe upon insertion in the sample were fitted to an approximate solution of the heat conduction equation, and the values of two parameters in that solution were determined. Using the values of these parameters, the thermal conductivity and thermal diffusivity of each sample were determined. The specific heat of each sample was estimated from the definition of thermal diffusivity. These thermophysical properties were expressed as a function of concentration and temperature.     

Preparation of samples for proficiency testing of pesticide residue analysis in processed foods

To conduct proficiency testing for the analysis of pesticide residues in processed foods, fortified samples of retort curry and pancake were examined. In the case of retort curry, heating and mixing were necessary at the time of preparation to provide a homogenous analytical sample. A mixture of 4 carbamates and 11 organophosphorus pesticides was spiked and 14 of them showed consistent results in the samples. In the case of pancake, 10 kinds of pesticides were added to the pastry. The prepared pastry was them cooked. The relative concentrations of most of the pesticides in the pancake were not affected and all the pesticides showed consistent results in the samples. These results showed that the two tested samples were suitable for proficiency testing.     

Next target for food hydrocolloid studies: Texture design of foods using hydrocolloid technology

Texture is important in terms of both food palatability and the safety of eating. Recently, the importance of texture has been emphasized for the development of nursing-care foods, including dysphagia foods, in recent aged society, where the number of patients with mastication and swallowing difficulties is increasing. Texture design of these food products is now one of the most important tasks in the food industry in Japan. Texture of these food products should be optimized by modulating viscoelasticity using hydrocolloids so that they can easily transform to ‘ready-to-swallow’ bolus during oral processing. This article reviews the importance of texture as an essential attribute of foods and also the usefulness of hydrocolloids as an ingredient to modify and control food texture. The article also covers recent trials by the author’s research team on bolus rheology and in vivo acoustic analysis. The trials are to find some objective parameters describing the mastication and swallowing eases as an alternative to conventional bulk rheology and subjective sensory analysis.     

Four-base codon-mediated incorporation of non-natural amino acids into proteins in a eukaryotic cell-free translation system

Various four-base codons have been shown to work for the introduction of non-natural amino acids into proteins in an Escherichia coli cell-free translation system. Here, a four-base codon-mediated non-natural mutagenesis was applied to a eukaryotic rabbit reticulocyte cell-free translation system. Mutated streptavidin mRNAs containing four-base codons were prepared and added to a rabbit reticulocyte lysate in the presence of tRNAs that were aminoacylated with a non-natural amino acid and had the corresponding four-base anticodons. A Western blot analysis of translation products indicated that the four-base codons CGGU, CGCU, CCCU, CUCU, CUAU, and GGGU were efficiently decoded by the aminoacyl-tRNAs having the corresponding four-base anticodons. In contrast, the four-base codons AGGU, AGAU, CGAU, UUGU, UCGU, and ACGU were not decoded. The stop codon-derived four-base codons UAGU, UAAU, and UGAU were found to be inefficient, whereas the amber codon UAG and opal codon UGA were efficient for the incorporation of non-natural amino acids. The application of the expanded genetic code in a eukaryotic cell-free system opens the possibility of a four-base codon-mediated incorporation of non-natural amino acids into proteins in living eukaryotic cells.     

Spatial distribution of submarine groundwater discharge and associated nutrients within a local coastal area

To understand the local-scale distribution of submarine groundwater discharge (SGD) and dissolved nutrients, a multiple-detector (222)Rn monitoring survey was undertaken along the Mt. Chokai volcanic coast in northern Japan. The surveys revealed that the highest SGD (calculated to be 6.2 × 10(4) m(3) d(-1), within an area of 2 × 10(4) m(2)) with the greatest nutrient fluxes (sum of NO(3)(-), NO(2)(-), and NH(4)(+) (DIN): 9.2 × 10(2) mol d(-1); PO(4)(3-) (DIP): 56 mol d(-1)) is present at the edge of the youngest volcanic lava flow in the area. Recharged groundwater transports nutrients through porous volcanic flows and discharges as SGD near shore. Our results demonstrate that the spatial distribution of SGD in the study area is closely regulated by the local geology and topography. Furthermore, we show that continuous (222)Rn monitoring with a multidetector system at boat speeds of 1-2 knots provides details at a scale one order of magnitude greater than has been reported previously. In addition, the results of our study suggest that SGD-borne DIP may play an important role in the important local oyster production.     

Hypotensive effect of a sweetpotato protein digest in spontaneously hypertensive rats and purification of angiotensin I-converting enzyme inhibitory peptides

We have investigated angiotensin I-converting enzyme (ACE) inhibitory activity in an enzyme digest of sweetpotato protein, the antihypertensive effect of the digest in spontaneously hypertensive rats (SHR), and the identification of an ACE inhibitory peptide. Protein was prepared from squeezed juice of sweetpotato by isoelectric focusing precipitation. Three kinds of proteases were selected for effective protein digestion. The digest, sweetpotato peptide (SPP), exhibited strong ACE inhibitory activity (IC₅₀: 18.2 μg/ml). SPP was orally administered by gavage to SHR at a dose of 100 mg/kg or 500 mg/kg. The systolic blood pressure and the diastolic blood pressure were measured at 0 (before administration), 2, 4, 8, and 24 h after administration. A dose-dependent decrease in systolic blood pressure in SHR was observed after oral administration of SPP. Significant differences between SPP-administered rats and control rats were observed 4 and 8 h after administration in the 500 mg/kg-administered group and 8 h after administration in the 100 mg/kg-administered group. Diastolic blood pressure also decreased in the SPP-administered groups, although the difference between SPP-administered rats and control rats was not significant. These results suggest that SPP may be useful in the prevention or treatment of hypertension. Peptides with ACE inhibitory activity were purified from SPP by absorption chromatography and preparative HPLC using an ODS column. The amino acid sequences of isolated peptides were I-T-P, I-I-P, G-Q-Y and S-T-Y-Q-T; their ACE inhibitory activities (IC₅₀) were 9.5 μM, 80.8 μM, 52.3 μM and 300.4 μM, respectively. In conclusion, I-T-P is a novel, strong ACE inhibitory peptide.     

Enzymatic hydrolysis of potato starches containing different amounts of phosphorus

The rapid hydrolysis of potato starches differing in phosphorus content, as well as sweet potato, cassava and yam starches, was accomplished by treatment of gelatinised starches with bacterial liquefying α-amylase at 50 °C for 1 h, followed by Bacillus licheniformis α-amylase at 55 °C up to 24 h, and then by glucoamylase at 40 °C for a further 24 h. Among the potato starches, the high-phosphorus starches showed higher starch resistant capacity than the medium-phosphorus starches, as well as other tuber and root starches. The hydrolysis rate of tuber and root starches was not greatly influenced by their amylose content and median granule size. Only glucose was detected in the almost completely hydrolysed tuber and root starch samples, indicating that the concomitant enzymes treatment could hydrolyse both the α-1,4 and α-1,6 linkages of the starches examined.