Biochemical Factors in the Maximal Growth of Tetrahymena

Dewey, Virginia C.

doi:10.2307/1538340

Cited by 16 publications

(10 citation statements)

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“…High levels of threonine can substitute for seine for release of inhibition but threonine is not a growth rate stimulator. 9. Comparisons are made between the data in the literature on vertebrates and tho'se reported here on Tetrahymena.…”

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confidence: 88%

Studies on the Biochemistry of Tetrahymena

Kidder

Dewey²

1947

Proc. Natl. Acad. Sci. U.S.A.

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Introduction.-Previous studiesl-7 have shown that the ciliated protozoan, Tetrahymena, has nutritional requirements very similar to animals in general. This similarity is especially striking in regard to amino acid requirements where the ten essential amino acids of the dog, man, etc., have.been shown to be essential also for Tetrahymena. Inasmuch as this protozoan possesses characteristics which make its culture in the absence of all other organisms relatively simple, it has been possible to investigate rather precisely the effects of various substances on its metabolism. Biochemical studies of this nature on other animals have been complicated and even vitiated by contaminating microorganisms of unknown synthetic abilities. This report deals with the growth responses of Tetrahymena to varying concentrations of the essential amino acids and to serine, and includes the results of tests involving availability of "natural and unnatural isomers. Experimental.-The organism used in this investigation was Tetrahymena geleii W grown in pure (bacteria-free) culture. The general techniques have been previously reported." 4, 8 Dose-response curves were constructed for each of the essential amino acids, and for the stimulatory amino acid, serine, by adding graded amounts to a base medium containing the other ten in the proportions found in gelatin (table 1, medium II) together with growth factors, dextrose and salts. The minimum concentrations which permitted optimum growth under these conditions were combined, but the nitrogen level was then inadequate so these amounts were increased fivefold (medium III, table 1). Dose-responses were then studied using these new proportions. This medium was also used in studies on the availability of unnatural isomers. The factor II preparations, which are necessary for the growth of this organism9' 10 were of two types. The one prepared from cerophyl, as

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confidence: 88%

Studies on the Biochemistry of Tetrahymena

Kidder

Dewey²

1947

Proc. Natl. Acad. Sci. U.S.A.

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“…The organism used in the present study was the ciliated protozoan Tetrahymena geleii W, which is the strain used in the previous studies on thiamine synthesis (Kidder and Dewey, 1942;1944). All work was done with pure (bacteria-free)…”

Section: Methodsmentioning

confidence: 99%

“…Very little growth occurred in the absence of the alfalfa extract and the addition of thiamine, riboflavin, pyridoxine, pantothenic acid, nicotinic acid, pimelic acid, z-inosital, uracil, or />-aminobenzoic acid either singly or in combination had no significant effect. Inasmuch as the heat-and alkali-treated alfalfa extract w r as certainly free of thiamine it was concluded that Tetrahymena could synthesize the thiamine required for its metabolic needs when supplied with Factor S. It was suggested that Factor S possibly acted as a catalyst necessary for the synthesis of the thiamine molecule.It was recognized that the alfalfa extract used contained Factors I and II (Dewey, 1941 ;1944) and we now know that the casein base contained Factor III (Kidder and Dewey, 1945a). This work was criticized by Hall and Cosgrove (1944) on the basis that the "vitamin-free" casein used for the base medium was not free of thiamine.…”

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confidence: 99%

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Studies on the Biochemistry of Tetrahymena. Iv. Amino Acids and Their Relation to the Biosynthesis of Thiamine

Kidder¹,

Dewey²

1945

The Biological Bulletin

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It was reported earlier (Kidder and Dewey, 1942) that two species of Tetrahymena were able to carry out the synthesis of thiamine, if provided with a substance found mainly in the leaves of plants. This substance was called Factor S and was found in highest concentration in alfalfa leaf meal but could not be demonstrated from materials of animal origin. Factor S was characterized by its solubility in water and alcohol (up to 75 per cent), insolubility in ether and acetone, stability to prolonged heat in the presence of either alkali or acid, and its stability to ultraviolet radiation. It was shown to be dialyzable through cellophane and not to be precipitated by the salts of heavy metals. It was shown that Tetrahymena gave optimal growth in a medium consisting of 'Vitamin-free" casein, salts and a heat-and alkali-treated water extract of alfalfa meal. Very little growth occurred in the absence of the alfalfa extract and the addition of thiamine, riboflavin, pyridoxine, pantothenic acid, nicotinic acid, pimelic acid, z-inosital, uracil, or />-aminobenzoic acid either singly or in combination had no significant effect. Inasmuch as the heat-and alkali-treated alfalfa extract w r as certainly free of thiamine it was concluded that Tetrahymena could synthesize the thiamine required for its metabolic needs when supplied with Factor S. It was suggested that Factor S possibly acted as a catalyst necessary for the synthesis of the thiamine molecule.It was recognized that the alfalfa extract used contained Factors I and II (Dewey, 1941 ;1944) and we now know that the casein base contained Factor III (Kidder and Dewey, 1945a). This work was criticized by Hall and Cosgrove (1944) on the basis that the "vitamin-free" casein used for the base medium was not free of thiamine. They reported growth of their strain of Tetrahymena in heat-and alkali-treated casein in the presence of thiamine and not in its absence. This criticism was shown to be invalid (Kidder and Dewey, 1944) when an extension of the earlier studies was carried out, using heat-and alkali-treated base media (casein, casein hydrolysate, gelatin, gelatin hydrolysate). It was then found that heat and alkali treatment of 1 Aided by grants from the Morgan Edwards Fellowship Fund,

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“…In a few years, other leaders, expanding the coverage of the whole area, emerged from laboratories around the world: in America, first Dewey, Elliott, Hetherington, Jahn, Lilly, Loefer, Pace, Phelps, Seaman, and Taylor, and then Blum, Browning, Buhse, Cameron, Conner, Ducoff, Dunham, Eichel, Frankel, Holz, Hutner, McCashland, McDonald, Miiller, Padilla, Scherbaum, Shrago, Slater, Stone, Sullivan, G. A. Thompson, Whitson, N. E. Williams, Wingo, Zimmerman, and others; in England, F. E. G. Cox, Curds, Kitching, Ryley, and Sleigh; in Czechoslovakia, Jirovec and colleagues; in Denmark, Zeuthen, Hamburger, Nilsson, Plesner, Rasmussen, and associates; in Japan, Watanabe and others. No historical survey should by-pass the importance of the early, thorough, and stimulating account of the biochemistry of Tetrahymena (essentially T. pyriformis) produced by the team of Kidder & Dewey (1951), a now classical work setting the stage for hundreds of subsequent studies-by the Amherst group and many others-of a nature more refined than ever before possible. The protozoologist Holz might well be mentioned here, also, as one of the principal leaders in tackling with ingenuity problems in the biochemistry and physiology of species of Tetrahymena other than T. pyriformis (see, for example, Holz, 1964, (1) The growth factor thioctic acid, known early in unidentified form (Dewey, 1944), was more precisely recognized and then chemically identified (at first under a variety of names, including protogen and a-lipoic acid) in several laboratories a few years later (see papers by Seaman, 1952;Slater, 1952;Stokstad et al, 1949). A lipoate of wide distribution in nature, it is required only by species of Tetrahymena and related ciliate genera.…”

Section: Early Work Landmarks and Leadershipmentioning

confidence: 99%