Nutritional requirements ofPrevotella sp. Isolated from the rumen of the goat

The nutritional requirements forPrevotella sp. 4PCCNB2 isolated from the rumen of a native goat in Korea and those of the ATCC 19189 strain isolated from the bovine rumen were investigated. The two strains grew well with ammonium sulfate as the sole added nitrogen source. However, neither a complex of amino acids nor casein hydrolysate effectively replaced ammonium sulfate. Biotin,p-aminobenzoic acid, and vitamin B₁₂ were essential to culture the ATCC 19189 strain. Unlike the ATCC 19189 strain, however, B₁₂ was only stimulatory for the growth of the 4PCCNB2 strain. The 4PCCNB2 strain grew well in the basal medium without an individual acid such as acetic acid or valeric acid. In contrast, either acetic or valeric acid was absolutely required for the growth of the ATCC 19189 strain.     

Nutritional requirements and biochemical activities of pineapple pink disease bacterial strains from Hawaii

Bacteria which cause pink disease of pineapple, identified on the basis of their nutritional and biochemical activities, were found to belong to three genera. These bacteria include the following species: Gluconobacter oxydans, Acetobacter aceti, and Erwinia herbicola. Several pink disease strains required one to three vitamins for growth. Both G. oxydans strains 303D and 180 required biotin, nicotinic acid, and pantothenic acid for growth; E. herbicola 189 required only nicotinic acid; however, A. aceti 295 was able to grow without any added supplements in glucose mineral salts medium. Optimal vitamin concentrations for maximal growth and optimal pH for the maximal number of generations per hour was established for a few pink disease strains.Journal Series Paper No. 2373 of the Hawaii Agricultural Experiment Station, Honolulu.     

Characterization of rat cecum cellulolytic bacteria.

Cellulose-degrading bacteria previously isolated from the ceca of rats have been characterized and identified. The most commonly isolated type was rods identified as Bacteroides succinogenes. These bacteria fermented only cellulose (e.g., pebble-milled Whatman no. 1 filter paper), cellobiose, and in 43 of 47 strains, glucose, with succinic and acetic acids as the major products. The only organic growth factors found to be required by selected strains were p-aminobenzoic acid, cyanocobalamine, thiamine, and a straight-chain and a branched-chain volatile fatty acid. These vitamin requirements differ from those of rumen strains of B. succinogenes, indicating the rat strains may form a distinct subgroup within the species. The mole percent guanine plus cytosine was 45%, a value lower than those (48 to 51%) found for three rumen strains of B. succinogenes included in this study. Cellulolytic cocci were isolated less frequently than the rods and were identified as Rumminococcus flavefaciens. Most strains fermented only cellulose and cellobiose, and their major fermentation products were also succinic and acetic acids. Their required growth factors were not identified but were supplied by rumen fluid.     

Batch production of propionic acid by immobilizedPropionibacterium sp

Whole cells ofPropionibacterium freudenreichii subsp.shermanii (two strains) were immobilized in a living state in 2 and 4% alginate gel and 2, 4 and 6% carrageenan gel. Production of propionic acid, acetic acid, and vitamin B₁₂ were examined. The best results were obtained in the fermentation with strains immobilized in 4% alginate gel when applied for the third time.     

Synthesis of 5-Alkyl-5-methyl-2-hydroxy-2-cyclopentenones

A hydrogen bacterium strain, N34, and its oxygen-resistant segregant strain, Y38, were subjected to a taxonomical study. Since both strains were capable of N₂-fixation, N₂-fixing facultative hydrogen autotrophs listed in “Bergey’s Manual of Systematic Bacteriology” were used for comparison. Both strains produced a water-insoluble carotenoid pigment, zeaxanthin dirhamnoside, indicating that both should be classified into the genus Xanthobacter. Then, the differential characteristics of the two species of the genus Xanthobacter, X. autotrophicus and X. flavus, were investigated as to both strains. The vitamin requirement, the sensitivity to oxygen under autotrophic conditions, the inducibility of hydrogenase, the substrate range of carbohydrates and N₂-fixing growth characteristics of both strains were almost completely opposite to those of X. flavus. Moreover, both strains coincided exactly with X. autotrophicus in morphological and other physiological characteristics. From these results both strains were identified as Xanthobacter autotrophicus.     

OBSERVATIONS SUR LES BESOINS EN VITAMINES DES DESMIDIÉES (CHLOROPHYCÉES-ZYGNEMATALES)1

The minimal nutrient requirements of 61 strains of desmids belonging to 18 different genera were investigated within 16 axenic cultures and 45 contaminated cultures. Thirty-nine strains were autotrophic: they included all 30 strains belonging to the genera Cosmarium, Staurastrum, and Micrasterias. Twenty-two strains were auxotrophic; they included all 14 strains belonging to the genus Closterium. Of the 22 auxotrophic strains, 13 required only vitamin B₁₂; the specific requirements of the other 9 strains were not determined.     

Isolation of Cobalt-Resistant Strains of Propionic Acid Bacteria,Potent Producers of Vitamin B12

A method of adaptation to cobalt nitrate at high concentrations allowed us to isolate 46 strains of propionic acid bacteria Propionibacterium acidipropionici, resistant to excessive amounts of Co²⁺ in the medium. Studies of these strains revealed cultures that were most potent in synthesizing vitamin B₁₂. The yield of vitamin B₁₂ was increased 3 times, compared to parent strains.     

The Physiology of Astrephomene gubernaculifera1

SYNOPSIS. A comparative physiologic study of 4- and 7-chromosome strains of Astrephomene gubernaculifera was done. The vitamins p-aminobenzoic acid, nicotinamide, biotin, thiamin HCl and vitamin B₁₂ were tested for their ability to support growth. Only vitamin B₁₂ was required for active growth altho the presence of thiamin HCl was necessary to produce structurally typical colonies. Astrephomene will not grow in the absence of an exogenous source of carbon. Of the 36 carbon sources tested, only pyruvate, butyrate, succinate and acetate permitted active growth. Sodium acetate was the best. Strains grew within the initial pH range of 5.0–7.5. At pH values above 8.0 growth declined rapidly. When media were buffered with TES (N-tris [hydroxymethyl]methyl-2-aminoethane-sulfonic acid) at an initial pH of 6.8 growth was enhanced. The organism grew at 15–40 C. Growth in the dark was slightly less than that in the light.     

Formation of Desthiobiotin from Oleic Acid by Brevibacterium sp.

Microbial formation of biotin-vitamers from oleic acid was investigated. Many strains of bacteria which were able to utilize oleic acid as a sole carbon source were isolated from soils and other natural materials. Among these bacteria, some strains formed a biotin-vitamer from oleic acid in the culture broth during the cultivation. The vitamer was purified from the culture broth of strain No. 23, and identified as desthiobiotin by chromatographical and biological methods.

From the results of investigation on the taxonomical characteristics, the bacterial strain No. 23 was assumed to be Brevibacterium sp.     

The Production of α-Ketoglutaric Acid from Salicylic Acid by Pseudomonas sp.†

Metabolites from salicylic acid by microorganisms were investigated. About eighty strains of bacteria which were able to utilize salicylic acid as a sole source of carbon were isolated from soil and other natural sources.

Among these bacteria, several strains produced a large amount of keto acids in the culture fluid during the cultivation. The acid was isolated from the culture fluid of strain K 102 in crystalline form. The crystal was identified as α-ketoglutaric acid by physicochemical methods. From the taxonomical studies, the isolated bacterial strains K 102 and K 362 were assumed to be Pseudomonas sp.     

Production of vitamin B12 by immobilized cells of a propionic acid bacterium

Summary The ability of immobilized cells of propionic acid bacteria to form vitamin B₁₂ has been investigated. Propionibacterium arl AKU 1251 having a considerable activity to produce the vitamin was selected as a test organism among six strains of propionic acid bacteria tested. The whole cells were entrapped with urethane prepolymers, photo-crosslinkable resin prepolymers or several other materials such as -carrageenan, agar or sodium alginate, and their vitamin B₁₂ productivity was compared. Based on the criteria of the convenience of preparation and the stability of the cell-entrapping gels, a hydrophilic urethane prepolymer, PU-9, was employed as gel material. Satisfactory vitamin B₁₂ production was obtained when 5–10 g of wet cells precultured to the late exponential growth phase were entrapped with 1 g of the prepolymer. Addition of a suitable amount of cobaltous ion and of 5,6-dimethyl benzimidazole to the culture medium was effective for the production of the vitamin by the immobilized cells. The repeated use of the immobilized cells was successfully achieved when a suitable amount of cells were entrapped and allowed the proliferation of cells inside gel matrices.     

Nutrition of Volvulina Playfair*

SYNOPSIS. Vitamin B₁₂, biotin and thiamine requirements of 10 strains of Volvulina steinii and 1 strain of V. pringsheimii were studied. Vitamin B₁₂ is required for growth of both species, thiamine stimulates growth slightly, and biotin has no discernible effect on growth. The minimum concentration of vitamin B₁₂ giving a growth response in V. steinii, strain SC-2, was 10^?8 g/ml, and maximum growth response was obtained with 1.1 × 10^?7 g/ml. An organic carbon source is required for growth of V. steinii but not of V. pringsheimii. Growth of V. steinii, strain SC-2, occurred in 20 of 21 carbon sources tested. Optimal growth with each carbon source was largely dependent upon pH. Except for pyruvate, acetate, and ethanol, carbon source utilization was light-dependent, and growth in ethanol was reduced in the dark. Isocitric lyase activity was detected in V. steinii grown on acetate medium.     

Systematic Separation and Identification of 13 Carbamate Pesticides in Their Mixture

About 2700 strains of various kinds of microorganisms isolated from soil were tested for phospholipase D activity. All of the 200 selected strains were actinomycetes which were divided into two groups on the basis of the isomeric form of diaminopimelic acid (A₂pm) present in the cell wall. Among 18 strains containing meso-A₂pm, two strains were selected on the basis of taxonomical characteristics and productivity of phospholipase D. These two strains were examined for microbiological characteristics and found to belong to the genera Actinomadura and Nocardiopsis.     

Global Analysis of the Genes Involved in the Thermotolerance Mechanism of Thermotolerant Acetobacter tropicalis SKU1100

Acetobacter tropicalis SKU1100 is a thermotolerant acetic acid bacterium that grows even at 42 °C, a much higher temperature than the limit for the growth of mesophilic strains. To elucidate the mechanism underlying the thermotolerance of this strain, we attempted to identify the genes essential for growth at high temperature by transposon (Tn10) mutagenesis followed by gene or genome analysis. Among the 4,000 Tn10-inserted mutants obtained, 32 exhibited a growth phenotype comparable to that of the parent strain at 30 °C but not at higher temperatures. We identified the insertion site of Tn10 on the chromosomes of all the mutant strains by TAIL (Thermal Asymmetric Interlaced)-PCR, and found 24 genes responsible for thermotolerance. The results also revealed a partial overlap between the genes required for thermotolerance and those required for acetic acid resistance. In addition, the origin and role of these thermotolerant genes are discussed.     

Effect of water soluble vitamins and their analogues on growth of candida albicans. III. Para-aminobenzoic acid,nicotinic acid,inositol and their analogues

Summary By using high concentrations of vitamins in biotin basal synthetic mineral medium,Candida albicans was shown to possess a partial dependency forp-aminobenzoic acid, nicotinic acid and inositol. Sodium gantrisin [N¹-3,4-dimethyl-5-isoxazolyl sulfanilamide)] was growth inhibitory in the absence ofp-aminobenzoic acid and its effect was reversed byp-aminobenzoic acid. Similarly, pyridine-3-sulfonic acid was growth inhibitory to the organism in the absence of nicotinic acid and its effect was reversed by nicotinic acid. Additions of biotins, thiamine, vitamin B₁₂, nicotinic acid,p-amino-benzoic acid and inositol to basal synthetic medium showed clear-cut growth stimulation for each vitamin. Parallel omission type experiments on the other hand, demonstrated a masking effect of one vitamin on another. Ascorbic acid, riboflavin, calcium pantothenate choline, folic acid and thioctic acid were without effect. A new procedure for the determination of microbial vitamin requirements was outlined.     

Growth promotion of <Emphasis Type="Italic">Xanthium italicum</Emphasis> by application of rhizobacterial isolates of <Emphasis Type="Italic">Bacillus aryabhattai</Emphasis> in microcosm soil

This study was conducted using rhizobacteria, which are able to exert beneficial effects upon plant growth in the infertile soil collected from barren lakeside areas. Four strains of plant growth promoting bacteria were isolated from the rhizosphere of a common wild plant, Erigeron canadensis. Isolated strains LS9, LS11, LS12, and LS15 were identified as Bacillus aryabhattai by 16S rDNA sequence analysis. B. aryabhattai LS9, LS11, LS12, and LS15 could solubilize 577.9, 676.8, 623.6, and 581.3 mg/L of 0.5% insoluble calcium phosphate within 2 days of incubation. Production of indole acetic acid, a typical growth promoting phytohormone auxin, by strain LS15 was 471.3 mg/L in 2 days with the addition of auxin precursor L-tryptophan. All the strains also produced other phytohormones such as indole butyric acid, gibberellins, and abscisic acid, and strain LS15 showed the highest production rate of gibberellin (GA₃), 119.0 μg/mg protein. Isolated bacteria were used in a microcosm test for growth of wild plant Xanthium italicum, which can be utilized as a pioneer plant in barren lands. Seed germination was facilitated, and the lengths of roots, and shoots and the dry weights of germinated seedlings after 16 days were higher than those of the uninoculated control plants. Root lengths of seedlings of X. italicum increased by 121.1% in LS11-treated samples after 16 days. This plant growth-promoting capability of B. aryabhattai strains may be utilized as an environmentally friendly means of revegetating barren lands, especially sensitive areas such as lakeside lands.     

Volatile Fatty Acid Requirements of Cellulolytic Rumen Bacteria

A gas chromatographic method was developed which could separate the isomers isovaleric and 2-methylbutyric acid. Subsequent analyses revealed that most commercially available samples of these acids were cross-contaminated; however, one sample of each acid was found to be pure by this criterion. The growth response of seven strains of cellulolytic rumen bacteria (three strains of Bacteroides succinogenes, three strains of Ruminococcus flavefaciens, and one strain of R. albus) to additions of isobutyric, isovaleric, 2-methylbutyric, valeric, and combinations of valeric and a branched-chain acid was determined. Strains of B. succinogenes required a combination of valeric plus either isobutyric or 2-methylbutyric acid. Isovaleric acid was completely inactive. Either isobutyric or 2-methylbutyric acid was required for the growth of R. albus 7. Strain C-94 of R. flavefaciens grew slowly in the presence of any one of the three branched-chain acids, but a combination of isobutyric and 2-methylbutyric acids appeared to satisfy this organism's growth requirements. None of the individual acids or mixtures of straight- and branched-chain acids allowed growth of R. flavefaciens strain C1a which would approach the response obtained from the total mixture of acids. Further work indicated that all three branched-chain acids were required for optimal growth by this strain, although isovaleric acid only influenced the rate of maximal growth. Either 2-methylbutyric or isovaleric acid allowed growth of nearly the same magnitude as that of the positive control for R. flavefaciens B34b. The presence of acetic acid had little influence on the rate or extent of growth of any of the strains except R. albus 7, for which the extent of growth was markedly increased. Determination of the quantitative fatty acid requirements for the three B. succinogenes strains indicated that 0.1 μmole of valeric per ml and 0.05 μmole of 2-methylbutyric per ml permitted maximal growth. However, with isobutyric acid as the branched-chain component, strains A3c and B21a required 0.1 μmole/ml in contrast to S-85 which exhibited optimal growth at the 0.05 μmole/ml level. By use of mixtures of isobutyric and 2-methylbutyric acids, good growth of C-94 was obtained at concentrations of 0.1 and 0.01 μmole/ml, respectively. About 0.3 μmole/ml of each acid was required for satisfactory growth of C1a.     

Nutritional interdependence betweenThermoanaerobacter thermohydrosulfuricus andClostridium thermocellum

Thermoanaerobacter thermohydrosulfuricus strain YM3 and Clostridium thermocellum strain YM4, obtained originally as a stable coculture, required yeast extract to grow separately. Cell-free broths of T. thermohydrosulfuricus strain YM3 and C. thermocellum strain YM4 monocultures replaced yeast extract in supporting the growth of strains YM4 and YM3, respectively. T. thermohydrosulfuricus strain YM3 produced vitamin B₆, B₁₂ analog(s), p-aminobenzoic acid and folic acid, which were required by C. thermocellum strain YM4. Likewise, strain YM4 produced niacin-active compound(s), thiamine, and methionine required by strain YM3. Received: 17 March 1995 / Accepted: 27 March 1995     

Transformation of Citric Acid to Acetic Acid,Acetoin and Diacetyl by Wine Making Lactic Acid Bacteria

A decrease in citric acid and increases in acetic acid, acetoin and diacetyl were found in the test red wine after inoculation of intact cells of Leuconostoc mesenteroides subsp. lactosum ATCC 27307. a malo-lactic bacterium, grown on the malate plus citrate-medium. Citric acid in the buffer solution was transformed to acetic acid, acetoin and diacetyl in the pH range of 2 to 6 after inoculation with intact cells of this bacterial species. It was concluded that citric acid in wine making involving malolactic fermentation, at first, was converted by citrate lyase to acetic and oxaloacetic acids, and the latter was successively transformed by decarboxylation to pyruvic acid which was subsequently converted to acetoin, diacetyl and acetic acid.

Both the activities of citrate lyase and acetoin formation from pyruvic acid in the dialyzed cell-free extract were optimal at pH 6.0. Divalent cations such as Mn²⁺, Mg²⁺, Co²⁺ and Zn²⁺ activated the citrate lyase. The citrate lyase was completely inhibited by EDTA, Hg²⁺ and Ag²⁺ . The acetoin formation from pyruvic acid was significantly stimulated by thiamine pyrophosphate and CoCl₂, and inhibited by oxaloacetic acid. Specific activities of the citrate lyase and acetoin formation were considerably variable among the six strains of malo-lactic bacteria examined. Some activities of irreversible reduction of diacetyl to acetoin were found in the cell-free extracts of four of the malo-lactic bacteria strains and the optimal pH was 6.0 for this activity of Leu. mesenteroides.     

Isolation and Identification of Acetic Acid Bacteria for Submerged Acetic Acid Fermentation at High Temperature

Industrial vinegar production by submerged acetic acid fermentation has been carried out using Acetobacter strains at about 30°C. To obtain strains suitable for acetic acid fermentation at higher temperature, about 1,100 strains of acetic acid bacteria were isolated from vinegar mash, soils in vinegar factories and fruits, and their activities to oxidize ethanol at high temperature were examined. One of these strains, No. 1023, identified as Acetobacter aceti, retained full activity to produce acetic acid in continuous submerged culture at 35°C and produced 45% of activity at 38°C, while the usual strain of A. aceti completely lost its activity at 35°C. Thus the use of this strain may reduce the cooling costs of industrial vinegar production.