Studies on Egg White Protein

The carbohydrate of ovomucoid was analyzed for components I, II, III and IV which were, fractionated by CMC-column chromatography. The total hexose content and the molar ratio of d-mannose to d-galactose (4:1) were identical in each component, but the d-glucosamine and sialic acid contents were found to be higher in components I and II (both are trypsin inhibitors) compared with components III and IV (both are apo-proteins of flavomucoid). The amino acid composition of each component of ovomucoid varied considerably. There were remarkable differences in the amino acid composition between components I and II, both had an antitryptic activity. The N-terminal amino acid of components I and II was alanine and in the case of components III and IV, threonine was found on the N-terminal. The free carboxylic residue of sialic acid was found to be responsible for the negative charge of ovomucoid, and its electrophoretic heterogeneity was reaffirmed by paper electrophoresis. It is evident from the ultracentrifugal analysis that the four components of ovomucoid have a similar molecular size.     

Studies on Egg White Proteins

Four components of ovomucoid were digested exhaustively and four kinds of glycopeptide corresponding to the four components were separated by gel filtration. Each glycopeptide was shown to be homogenious by paper chromatography and paper electrophoresis. Molar ratios of carbohydrate components of these glycopeptides varied to some extent but the amino acid compositions of these glycopeptides were essentially identical with each other with the exception of alanine. Aspartic acid and threonine were predominant amino acids in the all glycopeptides. It is most likely that the modes of linkages between polysaccharide and protein in individual ovomucoid I, II, III and IV are essentially the same, and that the carbohydrate moiety is linked to the protein via asparaginyl residue or the hydroxyl group of threonine, although the possibility of the linkages to glutamine and serine can not be excluded.     

Studies on the Foaming Property of the Chicken Egg White

The foaming power of the native ovalbumin has been found to be increased very much by various kinds of denaturation. The characters of its surface monolayer were also changed by denaturation procedure. From these resulte, the relation between the foaming power of the protein and the characters of its surface monolayer was discussed.     

Studies on the Foaming Property of the Chicken Egg White

The egg white was treated under various whipping conditions, and its foaminess measured. At the same time, the amounts of the coagulated proteins formed from each egg white and their constituent hexose were measured. From these results, discussions were made about the relation between the foaminess of the egg white and the amount of the coagulated proteins under various whipping conditions.     

Biotransformation of 2,4,6-Trinitrotoluene by Pure Culture Ruminal Bacteria

Twenty-one ruminal bacteria species were tested for their ability to degrade 2,4,6-trinitrotoluene (TNT) within 24 h. Butyrivibrio fibrisolvens, Fibrobacter succinogenes, Lactobacillus vitulinus, Selenomonas ruminantium, Streptococcus caprinus, and Succinivibrio dextrinosolvens were able to completely degrade 100 mg/L TNT, with <5% of the original TNT recovered as diaminonitrotoluene metabolites. Eubacterium ruminantium, Lactobacillus ruminis, Ruminobacter amylophilus, Streptococcus bovis, and Wolinella succinogenes were able to completely degrade 100 mg/L TNT, with 23–60% of the TNT recovered as aminodinitrotoluene and/or diaminonitrotoluene metabolites. Clostridium polysaccharolyticum, Megasphaera elsdenii, Prevotella bryantii, Prevotella ruminicola, Ruminococcus albus, and Ruminococcus flavefaciens were able to degrade 80–90% of 100 mg/L TNT. Desulfovibrio desulfuricans subsp. desulfuricans, Prevotella albensis, and Treponema bryantii degraded 50–80% of the TNT. Anaerovibrio lipolytica was completely inhibited by 100 mg/L TNT. These results indicate that a variety of rumen bacteria is capable of transforming TNT.     

Method for Radiorespirometric Detection of Bacteria in Pure Culture and in Blood

Methods are described for the detection of low numbers of bacteria by monitoring (14)CO(2) evolved from (14)C-labeled substrates. Cell suspensions are filtered with membrane filters, and the filter is then moistened with 0.1 ml of labeled medium in a small, closed apparatus. Evolved (14)CO(2) is collected with Ba(OH)(2)-moistened filter pads and assayed with conventional radioactivity counting equipment. The kinetics of (14)CO(2) evolution are shown for several species of bacteria. Fewer than 100 colony-forming units of most species tested were detected in 2 h or less. Bacteria were inoculated into blood and the mixture was treated to lyse the blood cells. The suspension ws filtered and the filter was placed in a small volume of labeled medium. The evolved (14)CO(2) was trapped and counted. A key development in the methodology was finding that an aqueous solution of Rhyozyme and Triton X-100 produced lysis of blood but was not detrimental to bacteria.     

Indigenous Bacteria and Fungi Drive Traditional Kimoto Sake Fermentations

Sake (Japanese rice wine) production is a complex, multistage process in which fermentation is performed by a succession of mixed fungi and bacteria. This study employed high-throughput rRNA marker gene sequencing, quantitative PCR, and terminal restriction fragment length polymorphism to characterize the bacterial and fungal communities of spontaneous sake production from koji to product as well as brewery equipment surfaces. Results demonstrate a dynamic microbial succession, with koji and early moto fermentations dominated by Bacillus, Staphylococcus, and Aspergillus flavus var. oryzae, succeeded by Lactobacillus spp. and Saccharomyces cerevisiae later in the fermentations. The microbiota driving these fermentations were also prevalent in the production environment, illustrating the reservoirs and routes for microbial contact in this traditional food fermentation. Interrogating the microbial consortia of production environments in parallel with food products is a valuable approach for understanding the complete ecology of food production systems and can be applied to any food system, leading to enlightened perspectives for process control and food safety.     

Studies on the Ventilation in Submerged Fermentations

Quantitative studies on the dissolution and dissociation of carbon dioxide in a cultured system were made. The inosine fermentation and the glutamic acid fermentation were employed for this study. According to the results obtained in this experiment, the quantity of dissociated carbonic acid in cultured liquid was given by Henderson-Hasselbalch’s equation with experimental pK′. The method for the direct determination of bicarbonate ion concentration was also investigated. The Warburg direct method gave a satisfactory result for this purpose.

By using the modified Severinghaus CO₂ electrode, the relationship between partial pressure of carbon dioxide in effluent gas and that in culturing system was investigated. Partial pressure of carbon dioxide in gas phase was almost equivalent to the average value of dissolved carbon dioxide tension in liquid phase for a given short time of the fermentation. The term of r_e was introduced in order to study the dynamic characteristics of carbon dioxide evolution in submerged fermentors. The dynamic characteristics of respiration in submerged fermentation was also studied by using biological r_ab and r_e.     

Studies on the Ventilation in Submerged Fermentations

As the basic research for the study of mechanism of ventilation in submerged fermentations, the formula of carbon dioxide in the liquid phase was investigated. Carbon dioxide in liquid phase was mainly classified into two groups. One was a gaseous form and the other was a dissociated form. Carbon dioxide was considered to be a weak acids when it dissociated into ionic form, and the dissociation of carbon dioxide was expressed in Henderson-Hasselbalch’s equation. The results in this study showed that those rules concerning the dissolution and dissociation of carbon dioxide held for a model liquid system.     

DNA Fingerprinting of Lactic Acid Bacteria in Sauerkraut Fermentations

Previous studies using traditional biochemical identification methods to study the ecology of commercial sauerkraut fermentations revealed that four species of lactic acid bacteria, Leuconostoc mesenteroides, Lactobacillus plantarum, Pediococcus pentosaceus, and Lactobacillus brevis, were the primary microorganisms in these fermentations. In this study, 686 isolates were collected from four commercial fermentations and analyzed by DNA fingerprinting. The results indicate that the species of lactic acid bacteria present in sauerkraut fermentations are more diverse than previously reported and include Leuconostoc citreum, Leuconostoc argentinum, Lactobacillus paraplantarum, Lactobacillus coryniformis, and Weissella sp. The newly identified species Leuconostoc fallax was also found. Unexpectedly, only two isolates of P. pentosaceus and 15 isolates of L. brevis were recovered during this study. A better understanding of the microbiota may aid in the development of low-salt fermentations, which may have altered microflora and altered sensory characteristics.     

Ecophysiological and Phylogenetic Studies of Nevskia ramosa in Pure Culture

During the last 100 years, the neuston bacterium Nevskia ramosa has been described several times. This bacterium forms conspicuous rosette-like microcolonies at the air-water interface. In this study, pure cultures of Nevskia ramosa were obtained for the first time, from a bog lake (strain Soe1, DSMZ 11499^T) and a freshwater ditch (strain OL1, DSMZ 11500). The isolates showed special adaptations to life in the epineuston. They formed hydrophobic surface films with a dull appearance. N. ramosa is sensitive to UV radiation but revealed a very effective photorepair mechanism. Exposure to light at a wavelength of 350 nm after UV treatment raised the number of surviving cells by several orders of magnitude. The isolates grew with a broad range of organic substrates. Surface films were formed only in the absence of combined nitrogen; however, nitrogenase activity was not detected. It appears that during growth at the air-water interface the cells benefit from trapping ammonia from the air. The G+C content of the DNA was 67.8 and 69.0 mol% for strains Soe1 and OL1, respectively. The slight difference was confirmed by enterobacterial repetitive intergenic consensus PCR. The 16S rRNA sequences revealed 99.2% similarity. Thus, both isolates belong to the same species. The phylogenetic analysis indicated that Nevskia is a member of the gamma-subclass Proteobacteria that has no known close relatives.Some morphologically conspicuous bacteria were observed in the 19th century but still have not been isolated in pure culture. In 1892, Famintzin (7) described Nevskia ramosa from the water surface of an aquarium in the botanical garden of St. Petersburg, Russia. The typical microcolonies consist of flat rosettes with a bush-like appearance on the water surface. The rosettes are colonies of dichotomously branched slime stalks with rod-shaped, slightly bent cells in the tips. The cells contain refractile globules, which were presumed to be ethereal oil (7), sulfur globules (12), or fat droplets (3). The slime stalks consist of polysaccharides (3) and sometimes appear to contain iron and aluminum encrustations (11).Enrichments of Nevskia-like cells in lake water supplied with lactate were described by Babenzien (1–4). He observed the following life cycle of N. ramosa. Young motile cells develop submersed, then adsorb to the water surface, lose the polar flagellum, and form a hyaline slime stalk on the concave side of the cell. When a cell multiplies by binary fission, branching of the stalk occurs. The resulting flat rosette can reach a size of 70 μm in diameter.Since pure cultures have not been available, little is known about the physiology, phylogeny, and ecology of Nevskia. It was assumed that Nevskia is oligocarbophilic (14). Tests with the nitrification inhibitor nitrapyrin gave no indications that the cells oxidize ammonia (16). N. ramosa was assumed to be related to the stalk-forming genera Caulobacter and Gallionella or to the sulfur-oxidizing Thiobacterium. In Bergey’s manual (4) N. ramosa was affiliated with the budding and/or appendaged bacteria.In addition to its conspicuous morphology, the typical habitat of N. ramosa prompted us to initiate the present investigation. The water-air interface is a very special environment, characterized by high surface tension and a relatively high hydrophobicity. Organic compounds and various typical bacteria are enriched in this zone. The living community is called the neuston (18, 21). Depending on whether they adsorb to the underside or the top of the water surface, organisms belong to the hyponeuston or epineuston, respectively. This habitat requires special adaptations with respect to adsorption, substrate uptake, and UV tolerance.In our study we have isolated N. ramosa in pure culture and carried out ecophysiological and phylogenetic characterizations. We found several adaptions to life in the epineuston in this interesting bacterium.