A chemically defined medium for Treponema strain PR-7 isolated from the intestine of a pig with swine dysentery

1972 ◽  
Vol 18 (7) ◽  
pp. 1073-1078 ◽  
Author(s):  
Robert M. Smibert ◽  
Raymond L. Claterbaugh Jr

The nutrition of treponeme strain PR-7 isolated from the intestinal tract of a pig with swine dysentery was studied. The organism fermented arabinose, xylose, maltose, cellobiose, glucose, galactose, mannose, lactose, pectin, and starch. The end products of fermentation of glucose were major amounts of succinic acid, moderate amounts of acetic and formic acid, and a trace of lactic acid and ethanol.Strain PR-7 required glutamate, aspartate, proline, leucine, methionine, arginine, valine, alanine, serine, lysine, glycine, threonine, isoleucine, phenylalanine, tyrosine, histidine, tryptophan, glutamine, asparagine, and spermine. The organism also required nicotinamide, folic acid, pyridoxal, thiamine, riboflavin, pantothenate, choline, α-lipoic acid, and biotin. The fatty acids isobutyrate, n-valerate, acetate, and pyruvate were needed as well as ammonium sulfate. A fermentable energy source such as glucose was necessary for growth, as well as carbon dioxide. Heme was also required and the organism was stimulated by vitamin B12, ascorbic acid, ornithine, and para-aminobenzoic acid. Heme could be replaced by catalase, myoglobin, and peroxidase.Ten other treponemes that were isolated from the intestines of normal pigs as well as from pigs with swine dysentery and had phenotypic characteristics similar to strain PR-7 grew in the defined medium.

2007 ◽  
Vol 73 (8) ◽  
pp. 2673-2681 ◽  
Author(s):  
Arno Wegkamp ◽  
Wietske van Oorschot ◽  
Willem M. de Vos ◽  
Eddy J. Smid

ABSTRACT The pab genes for para-aminobenzoic acid (pABA) biosynthesis in Lactococcus lactis were identified and characterized. In L. lactis NZ9000, only two of the three genes needed for pABA production were initially found. No gene coding for 4-amino-4-deoxychorismate lyase (pabC) was initially annotated, but detailed analysis revealed that pabC was fused with the 3′ end of the gene coding for chorismate synthetase component II (pabB). Therefore, we hypothesize that all three enzyme activities needed for pABA production are present in L. lactis, allowing for the production of pABA. Indeed, the overexpression of the pABA gene cluster in L. lactis resulted in elevated pABA pools, demonstrating that the genes are involved in the biosynthesis of pABA. Moreover, a pABA knockout (KO) strain lacking pabA and pabB C was constructed and shown to be unable to produce folate when cultivated in the absence of pABA. This KO strain was unable to grow in chemically defined medium lacking glycine, serine, nucleobases/nucleosides, and pABA. The addition of the purine guanine, adenine, xanthine, or inosine restored growth but not the production of folate. This suggests that, in the presence of purines, folate is not essential for the growth of L. lactis. It also shows that folate is not strictly required for the pyrimidine biosynthesis pathway. L. lactis strain NZ7024, overexpressing both the folate and pABA gene clusters, was found to produce 2.7 mg of folate/liter per optical density unit at 600 nm when the strain was grown on chemically defined medium without pABA. This is in sharp contrast to L. lactis strains overexpressing only one of the two gene clusters. Therefore, we conclude that elevated folate levels can be obtained only by the overexpression of folate combined with the overexpression of the pABA biosynthesis gene cluster, suggesting the need for a balanced carbon flux through the folate and pABA biosynthesis pathway in the wild-type strain.


1998 ◽  
Vol 64 (5) ◽  
pp. 1673-1679 ◽  
Author(s):  
Hua Wang ◽  
Weizhu Yu ◽  
Tim Coolbear ◽  
Dan O’Sullivan ◽  
Larry L. McKay

ABSTRACT A mutant of fast milk-coagulating (Fmc+)Lactococcus lactis subsp. lactis C2, designatedL. lactis KB4, was identified. Although possessing the known components essential for utilizing casein as a nitrogen source, which include functional proteinase (PrtP) activity and oligopeptide, di- and tripeptide, and amino acid transport systems, KB4 exhibited a slow milk coagulation (Fmc−) phenotype. When the amino acid requirements of L. lactis C2 were compared with those of KB4 by use of a chemically defined medium, it was found that KB4 was unable to grow in the absence of aspartic acid. This aspartic acid requirement could also be met by aspartate-containing peptides. The addition of aspartic acid to milk restored the Fmc+phenotype of KB4. KB4 was found to be defective in pyruvate carboxylase and thus was deficient in the ability to form oxaloacetate and hence aspartic acid from pyruvate and carbon dioxide. The results suggest that when lactococci are propagated in milk, aspartate derived from casein is unable to meet fully the nutritional demands of the lactococci, and they become dependent upon aspartate biosynthesis.


1973 ◽  
Vol 19 (10) ◽  
pp. 1319-1323 ◽  
Author(s):  
Brahma S. Sharma ◽  
Riaz-ul Haque

A synthetic medium, containing L-proline, glycine, L-arginine, L-cystine, L-glutamine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, nicotinamide, thiamine, glucose, sodium citrate, Na2HPO4, KH2PO4, (NH4)2SO4, MgSO4∙7H2O, FeSO4∙7H2O, and CaCl2∙2H2O, was developed. It supported growth and beta hemolysin production by the 681C strain of Staphylococcus aureus when the culture was incubated under carbon dioxide. L-Threonine, L-tyrosine, i-inositol, folic acid, riboflavin, pyridoxal HCl, choline Cl, and D-Ca pantothenate were added for obtaining good growth and toxin formation under air. L-Proline, L-glutamine, and L-cystine were the absolute requirements for the production of beta hemolysin under carbon dioxide. Carbon dioxide stimulated the production of beta hemolysin, but invariably resulted in lower cell yield.


2006 ◽  
Vol 72 (1) ◽  
pp. 319-326 ◽  
Author(s):  
Frederik Vaningelgem ◽  
Veerle Ghijsels ◽  
Effie Tsakalidou ◽  
Luc De Vuyst

ABSTRACT Citrate metabolism by Enterococcus faecium FAIR-E 198, an isolate from Greek Feta cheese, was studied in modified MRS (mMRS) medium under different pH conditions and glucose and citrate concentrations. In the absence of glucose, this strain was able to metabolize citrate in a pH range from constant pH 5.0 to 7.0. At a constant pH 8.0, no citrate was metabolized, although growth took place. The main end products of citrate metabolism were acetate, formate, acetoin, and carbon dioxide, whereas ethanol and diacetyl were present in smaller amounts. In the presence of glucose, citrate was cometabolized, but it did not contribute to growth. Also, more acetate and less acetoin were formed compared to growth in mMRS medium and in the absence of glucose. Most of the citrate was consumed during the stationary phase, indicating that energy generated by citrate metabolism was used for maintenance. Experiments with cell-free fermented mMRS medium indicated that E. faecium FAIR-E 198 was able to metabolize another energy source present in the medium.


1958 ◽  
Vol 4 (5) ◽  
pp. 525-528 ◽  
Author(s):  
H. E. Swim ◽  
R. F. Parker

The results of these studies demonstrate that carbon dioxide is required for the growth and maintenance of strains of fibroblasts derived from human tissues, strains FS4-705 and U12-705, from mouse tissue, strain L-705, and from rabbit tissues, strains RM3-56, RS1-56, and RT-56 in a chemically defined medium containing phosphite buffer in place of bicarbonate and supplemented with dialyzed serum and dialyzed embryo extract. Under these conditions, the cells fail to proliferate at a significant rate and begin to degenerate within 5 to 10 days when the flasks are not stoppered. Sufficient carbon dioxide is produced by the cells to promote growth as indicated by the fact that maximal proliferation is obtained in the same phosphite media when stoppered flasks are employed. With the exception of RS1-56, all the remaining strains tested can be propagated serially in open flasks containing phosphite medium prepared with whole serum and embryo extract. The rate of growth under these conditions, however, is only one-half to one-third that obtained in stoppered flasks containing phosphite medium or the conventional bicarbonate medium.


1970 ◽  
Vol 16 (9) ◽  
pp. 809-815 ◽  
Author(s):  
Horace J. Daniels

The effect of known precursors and cofactors on vitamin B12 production has been studied in a basal, chemically defined medium composed of sucrose, glutamic acid, and mineral salts. Only a carbon source, betaine, and Co2+ were found to be essential for vitamin B12 production. Known precursors such as methionine, δ-aminolevulinic acid, succinic acid, and 1-amino-2-propanol had no effect. Glycine was inhibitory to growth. Oxalic acid, lactic acid, 5,6-dimethylbenzimidazole, and Mo2+ stimulated vitamin B12 biosynthesis. Of special interest is oxalic acid, which at a level of 0.04% w/v increased vitamin production about 20%, and with washed cells could replace glutamic acid, which had been demonstrated to be essential for both growth and vitamin B12 production. When use of oxalate was followed, it was found that this acid disappeared during the period of rapid growth and vitamin production and then reappeared during the final stages of the fermentation cycle.


1965 ◽  
Vol 11 (6) ◽  
pp. 1009-1019 ◽  
Author(s):  
Lillian V. Holdeman ◽  
Louis Ds. Smith

Clostridium botulinum type F was grown in a chemically defined medium containing 17 amino acids, 11 vitamins, glucose, and inorganic salts. The nutritional requirements were determined using single-omission test media. Arginine, tryptophan, tyrosine, valine, biotin, thiamin, and possibly methionine were essential nutrients. Growth was stimulated by glycine, isoleucine, phenylalanine, and para-aminobenzoic acid. Toxin was present in supernatant fluids from all synthetic medium cultures in which there was marked growth. In general, toxicity of synthetic medium cultures was about 1/10 that of complex medium cultures.Toxin and precursor appear to be formed intracellularly, for both were released by rupturing young cells with sonic vibration. Protoxin could be activated by treatment with trypsin.


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