Quantitative Chromatographic Estimation of α-Amino-Acids

Naftalin, L.

doi:10.1038/161763a0

Cited by 67 publications

(16 citation statements)

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“…Disappearance of L-glutamic acid and L-histidine from the medium was followed by the paper chromatographic procedure of Naftalin (41) and by means of the automatic analytical Beckman Spinco amino acid analyzer (model 120C) (58,59). An aliquot of the culture filtrate (5 ml, adjusted to pH 2.5 to 3.0) was passed through a Dowex 50W column (1 by 6 cm, H+ form, 200 to 400 mesh), and the amino acids were eluted with 2 N NH40H (34).…”

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confidence: 99%

Development of a Chemically Defined Medium for the Synthesis of Actinomycin D by Streptomyces parvulus

Williams

Katz

1977

Antimicrob Agents Chemother

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A chemically defined medium, consisting of D-fructose, L-glutamic acid, L-histidine, K2HPO4, MgSO4 * 7H2O, ZnSO4 * 7H2O, CaCl2 * 2H2O, FeSO4 * 7H20, CoC12 -6H2O, and deionized water, was developed for synthesis of high yields (500 to 600 ,ug/ml) of actinomycin D by Streptomyces parvulus. Under these nutritional conditions, growth and actinomycin formation did not follow a typical trophophase-idiophase pattern. The amino acids appeared to have a sparing action on the utilization ofD-fructose which was slowly and incompletely metabolized during mycelium development and antibiotic production. A significant repression of actinomycin synthesis by S. parvulus was observed when Dglucose (0.01 to 0.25%) was added to the culture medium. The repression was not due to a decline in the pH of the medium during glucose catabolism.Because of our continued interest in the mechanism of directed biosynthesis (30), we have turned our attention to the metabolic activities of Streptomyces parvulus which synthesizes virtually the single antibiotic component, actinomycin D (40). Although the medium described previously (16, 32) proved satisfactory for the production of actinomycin mixtures by Streptomyces antibioticus, the synthesis of actinomycin D by S. parvulus, under the same conditions, was rather poor and extremely variable. Studies were, therefore, initiated to establish the nutritional factors that would support growth and consistently high yields of the antibiotic by this organism. This report describes a series of experiments that have culminated in the development of a chemically defmed medium in which production of high titers of actinomycin D was achieved.MATERIALS AND METHODS Organism and conditions of cultivation. S. parvulus (ATCC 12434) was maintained on slants of glucose-yeast extract-malt extract agar medium (26). Incubation was carried out for 3 to 5 days at 30°C until gray spores had developed; slants were then kept at 40C for further use.For preparation of vegetative inoculum, spores were scraped gently from the surface of a slant(s) with 2.5 ml of NZ-Amine medium (31), and 2-ml quantities of the spore suspension were inoculated into a 250-ml Erlenmeyer flask containing 100-ml amounts of NZ-Amine medium. After incubation on a gyratory shaking incubator (New Brunswick Scientific Co., Inc., New Brunswick, N.J.) for 48 h at 3000, the mycelium was harvested by centrifugation (30 min at 9;000 rev/min), washed twice with and then suspended in 100 ml of saline as described previously (31, 34). Aliquots (3 ml) ofthe suspension served as inoculum for each 250-ml Erlenmeyer flask containing 100 ml of chemically defined medium.Basal medium for nutritional studies. The medium (D-galactose, 10 g; 1-glucose, 1.0 g; L-glutamic acid * HCI, 2.5 g; MgSO4 * 7H20,25 mg; ZnSO4 *7H20, 25 mg; CaCl2 *2H20, 25 mg; FeSO4 7H20, 25 mg; deionized water, 1,000 ml; pH 7.1) routinely utilized for actinomycin production with S. antibioticus and Streptomyces chrysomallus (16, 33, 34, 60) was employed initially for the studies with S. parvulus. Ca...

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Development of a Chemically Defined Medium for the Synthesis of Actinomycin D by Streptomyces parvulus

Williams

Katz

1977

Antimicrob Agents Chemother

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“…The concentration of actinomycin was determined spectrophotometrically (23). Determination of amino acids was accomplished with the Beckman Spinco automatic amino acid analyzer (model 120C) (38,39) or by a modification of the paper chromatographic procedure described by Naftalin (23,29). Optical configuration of amino acids was established with dialyzed prepa-226 YAJIMA, MASON, AND KATZ rations of L-or 1-amino acid oxidase (23,25,30).…”

Section: Methodsmentioning

confidence: 99%

Biogenetic Origin of the d -Isoleucine and N -Methyl- l -Alloisoleucine Residues in the Actinomycins

Yajima

Mason

Katz

1976

Antimicrob Agents Chemother

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Studies with 14 C-labeled isoleucine stereisomers have established that l -alloisoleucine, d -alloisoleucine, and d -isoleucine may function as precursors for the biogenesis of d -isoleucine and N -methyl- l -alloisoleucine residues in actinomycin. l -[ 14 C]isoleucine appears to be employed chiefly for d -alloisoleucine (and N -methylisoleucine [?] formation); however, its role in the biosynthesis of d -isoleucine and N -methylalloisoleucine remains unclear. The potential pathway of biosynthesis of d -isoleucine and N -methyl- l -isoleucine is discussed.

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“…ou por cromatograma paralelo; c) revelando o cromatograma v cortando as manchas obtidas; eluindo e determinando a intensidade da cor eluida; d) nedindo por densitometria a intensidade da cor da mancha sobre o papel (NAFTALIN, 1948; BULL, HAHN e BAPTIST, 1949; ROCKLAND, 1 Trabalho realizado com auxílio da Fundação de Amparo à Pesquisa do Estado de S. Paulo. Recebido para publicação em 11/9/67.…”

Section: Introductionunclassified

Determinação quantitativa de putrescina por cromatografia em papel de filtro

Crocomo¹,

Rossi²

1967

An. Esc. Super. Agric. Luiz de Queiroz

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RESUMOO presente trabalho descreve o desenvolvimento de um método de determinação quantitativa da amina putrescina por cro¬ matografia em papel de filtro seguida de espectrofotometria. 80-85% de putrescina adicionada a extratos de plantas pode ser recuperada.O espectro de absorção do produto de reação entre putrescina e ninhidrina tem um pico máximo a 575 mu.O desenvolvimento da côr do produto de reação atinge um máximo em 20 minutos, a 65ºC e se mantém estável durante os 10 minutos seguintes. O produto colorido eluido do cromatograma é estável durante 3 horas. INTRODUÇÃOOs métodos de determinação cromatogrãfica quantitativa de compostos químicos que dão reação positiva com tricetohiirindeno hidratado (ninhidrina), como os aminoácidos» amidas e aminas» baseiam-se seja: a) na medida da ãrea da mancha sobre o papel de filtro; b) "eluindo" a ãrea ocupada peló composto no papel» localizada por fluorescencia ao U.V. ou por cromatograma paralelo; c) revelando o cromatograma v cortando as manchas obtidas; eluindo e determinando a intensidade da cor eluida; d) nedindo por densitometria a intensidade da cor da mancha sobre o papel (NAFTALIN, 1948; BULL, HAHN e BAPTIST, 1949; ROCKLAND, 1 Trabalho realizado com auxílio da Fundação de Amparo à Pesquisa do Estado de S. Paulo. Recebido para publicação em 11/9/67.

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Quantitative Chromatographic Estimation of α-Amino-Acids

Cited by 67 publications

References 3 publications

Development of a Chemically Defined Medium for the Synthesis of Actinomycin D by Streptomyces parvulus

Development of a Chemically Defined Medium for the Synthesis of Actinomycin D by Streptomyces parvulus

Biogenetic Origin of the d -Isoleucine and N -Methyl- l -Alloisoleucine Residues in the Actinomycins

Determinação quantitativa de putrescina por cromatografia em papel de filtro

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