Naoko Hamada-Sato

Effect of culture conditions on lactic acid production of Tetragenococcus species

Aims:  To investigate the effects of the salt concentration, incubation temperature and initial pH of the medium on the fermentative ability of the halophilic lactic acid bacteria, Tetragenococcus muriaticus and T. halophilus.

Actinomycetal Community Structures in Seawater and Freshwater Examined by DGGE Analysis of 16S rRNA Gene Fragments

The actinomycetal community structures in marine and freshwater environments (the Pacific Ocean, East China Sea, Tokyo Bay, and Arakawa River) were investigated by a culture-independent molecular method to clarify spatial and seasonal distributions. Deoxyribonucleic acid (DNA) was extracted from environmental water samples, and a community analysis was carried out on polymerase chain reaction-amplified 16S ribosomal DNA. The amplified DNA fragments were analyzed by denaturing gradient gel electrophoresis (DGGE) and nonmetric multidimensional scaling analysis, followed by sequencing analysis. The actinomycetal community structures were different at each station in the Pacific Ocean, the East China Sea, Tokyo Bay, and Arakawa River, and different populations predominated in each area. There were vertical variations in actinomycetal communities in the Pacific Ocean and East China Sea between the surface and 100-m depth, but communities were similar from 200- to 1,000-m depths. There were also distinct seasonal variations in communities in Tokyo Bay. Phylogenetic analysis of DNA fragments recovered from DGGE bands revealed that most of the predominant actinomycetal strains were uncultured and were quite different from well known culturable strains, such as the Streptomyces, Micromonospora, Microbispora, Salinispora, and Actinoplanes groups. These results suggest that the marine environment is an attractive target for discovering new actinomycetal populations producing bioactive compounds and that sampling depth and season are important considerations for isolating various populations effectively.

Quantification of major allergen parvalbumin in 22 species of fish by SDS-PAGE.

Fish is an important causative material of food allergy. Although the allergenicity of fish is considered to correlate with the content of parvalbumin, the major fish allergen, available information about the parvalbumin content in fish is limited. In this study, a simple and reliable quantification method for fish parvalbumin by SDS-PAGE was first established. Application of the SDS-PAGE method to 22 species of fish revealed a marked variation in parvalbumin content among fish. Furthermore, the parvalbumin content was found to be higher in dorsal white muscle than in ventral white muscle, in rostral part of white muscle than in caudal part of white muscle and in white muscle than in dark muscle. IgE reactivity of fish was roughly proportional to parvalbumin content. Interestingly, large-sized migratory fish, such as salmon, swordfish and tuna, were commonly very low in both parvalbumin content and IgE reactivity.

Isolation, characterization, and utilization of γ-aminobutyric acid (GABA)-producing lactic acid bacteria from Myanmar fishery products fermented with boiled rice

Abstractγ-Aminobutyric acid (GABA)-producing lactic acid bacteria (LAB) were isolated from four types of Myanmar traditional fermented fishery products with boiled rice. All of them belonged to the genus Lactobacillus, and comparison of the effects of these representatives on GABA accumulation in fermented fishery products with boiled rice revealed that Lactobacillus farciminis D323 is the most effective strain as a starter culture. These results may contribute to the development of traditional fermented fishery products with functional properties. In addition, this study is the first to show in detail the distribution of GABA-producing LAB in Southeast Asian fermented fishery products.

Innovative food processing technology using ohmic heating and aseptic packaging for meat

Since the Tohoku earthquake, there is much interest in processed foods, which can be stored for long periods at room temperature. Retort heating is one of the main technologies employed for producing it. We developed the innovative food processing technology, which supersede retort, using ohmic heating and aseptic packaging. Electrical heating involves the application of alternating voltage to food. Compared with retort heating, which uses a heat transfer medium, ohmic heating allows for high heating efficiency and rapid heating. In this paper we ohmically heated chicken breast samples and conducted various tests on the heated samples. The measurement results of water content, IMP, and glutamic acid suggest that the quality of the ohmically heated samples was similar or superior to that of the retort-heated samples. Furthermore, based on the monitoring of these samples, it was observed that sample quality did not deteriorate during storage.

Improvement of Fish Freshness Determination Method by the Application of Amorphous Freeze-Dried Enzymes

Alkaline phosphatase (ALP), nucleoside phosphorylase (NP), and xanthine oxidase (XOD) were used in a colorimetric method for evaluation of fish freshness based on the Ki value. Two enzyme mixtures, NP-XOD and ALP-NP-XOD, were prepared with a color developing agent, and stabilities of the enzymes were improved by freeze-drying with glass-forming additives, i.e., sucrose and sucrose-gelatin. As a result, a linear relationship was obtained between the Ki values determined by the developed colorimetric method and a conventional high-performance liquid chromatography with a high correlation coefficient of 0.997. All enzyme samples containing the additive(s) were amorphous, and higher enzymes activities were maintained compared to those freeze-dried without an additive. Sucrose-gelatin/enzyme mixtures showed higher glass transition temperature; consequently, the enzymes were better stabilized than the sucrose/enzyme formulations. Using the sucrose-gelatin/enzyme mixture, Ki values of fish meat could be accurately determined even after 6-month storage of the dried enzymes at 40 °C.

Study of the cross-reactivity of fish allergens based on a questionnaire and blood testing

BACKGROUND Parvalbumin and collagen have been identified as cross-reactive allergens for fish allergies. Although doctors realize that various fish elicit allergies, the targets of food allergen labeling laws were only mackerels and salmons in Japan and mackerels in South Korea. This study aimed to reveal the causative species for fish allergy via questionnaires and blood tests. METHODS Questionnaire research was conducted in Japan via the internet concerning allergies for fish-allergic patients or their family members. Next, IgE reactivities and cross-reactivities of 26 fish species were analyzed using sera obtained from 16 Japanese patients who were allergic to fish parvalbumin or collagen by enzyme-linked immunosorbent assay (ELISA) and inhibition ELISA. RESULTS Questionnaire research revealed that 88% patients cannot eat mackerel and salmon in addition to other fish. In addition, 85% respondents were not satisfied with the current food allergen labeling law. In ELISA analyses, we clarified that pooled serum obtained from patients with fish parvalbumin-specific allergies exhibited IgE reactivity to the extracts of most fish species, and pooled serum obtained from patients with fish collagen-specific allergies displayed IgE reactivity to the extracts of all types of fish. Inhibition ELISA experiments revealed cross-reactivities of parvalbumin or collagen to extracts from all fish tested. CONCLUSIONS Most patients with fish allergies displayed allergic symptoms following the intake of various fish species. In addition, fish parvalbumin and collagen were causative factors of fish allergy and were highly cross-reactive fish panallergens. Therefore, current laws should be revised in Japan and South Korea.

Equations for spectrophotometric determination of relative concentrations of myoglobin derivatives in aqueous tuna meat extracts

The percentage of metmyoglobin (%metMb) in aqueous meat extracts of bigeye and bluefin tuna and beef samples were estimated using previously reported equations derived from the absorption spectra of horse Mb. The results demonstrate that in an aqueous extract, the difference in %metMb estimated by the different equations was negligible for beef samples. Conversely, in an aqueous tuna extract, different %metMb values were obtained with the different equations. The discrepancy in the tuna sample results might be due to differences in absorption spectra for horse and tuna Mb. Therefore, a new set of equations derived from the absorption spectra of bigeye tuna Mb, reported by Matsuura and Hashimoto (1955), was established. The accuracy of the proposed equations was compared with the cyanmetmyoglobin (cyanmetMb) method. The results show that the total Mb concentrations estimated by our proposed equations were in good agreement with the results obtained by the conventional cyanmetMb method (R(2)=0.984). Therefore, the new set of proposed equations is valid for the spectrophotometric determination of the relative proportions of Mb derivatives and total Mb concentration in aqueous tuna meat extracts.

Reduction in IgE reactivity of Pacific mackerel parvalbumin by heat treatment

Parvalbumin, a major fish allergen, has been reported to be highly thermostable. However, little is known as to whether parvalbumin is stable at more than 100°C. Thermostability of the Pacific mackerel parvalbumin was examined by subjecting heated (20-140°C) muscle extracts to SDS-PAGE, western blotting and ELISA. As judged by SDS-PAGE and western blotting with the anti-parvalbumin antiserum recognizing the primary structure, the parvalbumin was not degraded even under severe heating conditions. However, western blotting analysis with the monoclonal antibody recognizing the stereoscopic structure revealed that the parvalbumin undergoes conformational changes in a heating load-dependent manner. Importantly, the IgE reactivity of the parvalbumin determined by ELISA using patient sera was also reduced in a heating load-dependent manner; complete loss of IgE reactivity was induced by heating at 140°C. This study showed that the allergenicity of the Pacific mackerel parvalbumin is considerably less thermostable than assumed for other fish parvalbumins.

Degradation of Ferric Chelate of Ethylenediaminetetraacetic Acid by Bacterium Isolated from Deep-Sea Stalked Barnacle

Twenty strains of marine bacteria that degrade ferric chelate of ethylenediaminetetraacetic acid (Fe-EDTA) were isolated from among 117 strains collected from a marine environment. Among them strain 02-N-2, which was isolated from stalked barnacle collected from the deep sea in the Indian Ocean, had the highest Fe-EDTA degradation ability and was selected for further study. The strain showed high Fe-EDTA degradation ability at different seawater concentrations. In addition, the intact cells of this strain had the ability to degrade such metal-EDTAs as Ca, Cu, and Mg. The strain was an aerobic, gram-variable, rod-shaped organism. The results of various taxonomic studies revealed that the strain had significant similarity to Bacillus jeotgali JCM 10885T, which was isolated from a Korean traditional fermented seafood, Jeotgal.