Biotin Requirement Research Articles

Identification of Bacillus subtilis NRRL B-3275 as a strain of Bacillus pumilus.

The physiological and biochemical properties of a species of Bacillus previously identified as B. subtilis NRRL B-3275 (B-3275) were compared with those of seven strains of B. pumilus and five strains of B. subtilis. The biotin requirement of B-3275, its inability to hydrolyze starch, and its failure to reduce nitrate indicate that the organism is more closely related to the B. pumilus strains than to those of B. subtilis. Hybridization of deoxyribonucleic acid (DNA) from B-3275 with that of the strains of B. pumilus showed a binding efficiency (compared with the homologous reaction) of 58 to 99%, depending on the strain. Hybridization with the DNA from any of the strains of B. subtilis did not exceed 24%. DNA from B-3275 was unable to transform two amino acid auxotrophic markers to prototrophy in a highly competent strain of B. subtilis 168. We conclude that B-3275 is a strain of B. pumilus which we designate as B. pumilus NRRL B-3275.

The requirement for biotin in mouse fibroblast L-cells cultured on serumless medium

Mouse L-cells, Clone 929, were adapted to growth on a chemically defined medium, Waymouth's Medium MAP 954 1 , in the absence of serum. When biotin was excluded in the preparation of this medium, the resulting medium proved to be incapable of sustaining continuous cell proliferation: A biotin requirement was therefore demonstrable in these cells within a three week period.

Requirement of Turkey Poults for Biotin and Effect of Deficiency of Incidence of Leg Weakness in Developing Turkeys

SOON after the discovery of biotin its nutritional significance in chick and poult nutrition was investigated by Patrick et al. (1942) who observed that a deficiency resulted in perosis and a dermatitis that appeared first on the foot pads. Biotin requirement of poults was estimated to be five micrograms per day during the first four weeks and it was suggested that practical poult diets but probably not chick diets may be deficient in this vitamin. Evidence was obtained for intestinal synthesis of biotin in laying hens (Couch et al., 1948). Robblee and Clandinin (1953) suggested that biotin may be marginal in practical poult rations. Antibiotics were shown to reduce the requirement of the turkey poult for biotin by Slinger and Pepper (1954) in studies which indicated that supplemental biotin to practical rations was probably not necessary.Interest in the biotin needs of turkeys has increased greatly because of the appearance…

Taxonomy of the aerobic psuedomonads: Pseudomonas diminuta and P. vesiculare.

SUMMARY: Thirteen strains previously assigned to the species Pseudomonas diminuta and P. vesiculare were subjected to detailed characterization. Ten of these strains could be identified as P. diminuta and two as P. vesiculare. These two species can be readily distinguished by differences in their nutritional spectra, their growth factor requirements and their pigmentation. The DNA of the two strains of P. vesiculare contained 65.8 moles% guanine+cytosine (GC), and the DNA of the ten strains of P. diminuta from 66.3 to 67.3 moles%. One of the 13 strains examined had DNA of significantly lower GC content (62.2 moles%) and also differed in several phenotypic respects from both species; it is probably a monotypic representative of a third species, as yet unnamed. All the strains examined share a series of distinctive properties, which justify their recognition as a special subgroup of aerobic pseudomonads. The defining characters of this subgroup include: monotrichous flagellation, with flagella of very short wavelength; a requirement for pantothenate, biotin and cyanocobalamin; a limited range of carbon sources; production of acid from primary alcohols by all strains that can utilize alcohols; inability to denitrify or to use nitrate as a nitrogen source; and accumulation of poly-β-hydroxybutyrate as an intracellular reserve.

Nutritional requirements of mouse adrenal cortex tumor cells in culture

The minimal essential nutritional requirements for stable, highly differentiated clonal and non-clonal cultures of mouse adrenal cortex tumor cells were found to agree qualitatively with results reported earlier by Eagle for other mammalian cell lines which varied widely as to species and tissue origin. A requirement for tryptophan, biotin or vitamin B 12 could not be positively established in the presence of low concentrations of dialyzed horse serum. It was not possible to demonstrate a loss in function without a parallel loss in total cellular protein under the experimental conditions cited.

Effects of biotin on glutamate production and fatty acid composition in Bacillus cereus 14B22.

Bacillus cereus 14B22 exhibits a biochemical regulation by biotin. In glucose–salts or citrate–salts media, limiting amounts of biotin restrict cellular synthesis and induce synthesis of high yields of extracellular L-glutamic acid. With nonlimiting levels of biotin, abundant cellular synthesis occurs without the concomitant accumulation of glutamate. The biotin status of cells was not reflected in experiments with cell-free extracts. Citrate was converted to glutamate at comparable rates by extracts of biotin-sufficient and biotin-deficient cells. The biotin requirement for growth can be replaced with Tween 80 or with a mixture of Tween 40 and oleic acid. The yields of glutamate given by cultures of these "fatty acid-grown cells" were intermediate between those obtained with biotin-deficient and biotin-sufficient cells. Biotin deficiency also altered the fatty acid composition of the cell membrane. The proportion of straight-chain to branched-chain fatty acids is significantly smaller in low-biotin cells than in high-biotin cells. These changes did not affect cellular permeability to glutamate. Variations in the morphology of B. cereus 14B22 as influenced by biotin and the fatty acid composition of the medium are described.

Studies on a biotin requiring strain of Verticillium dahliae

Verticillium dahliae isolated from wilting Hedysarum coronarium was shown to have a marked requirement for bio tin. When it was inoculated on to/into normal Dox's agar/liquid medium it produced only a very thin, almost negligible, mat of abnormal hyphae, very sparse sporulation and hyaline microsclerotia, whereas on potato-dextrose agar (the potato tuber extract probably containing bio tin) growth was identical with that of a typical isolate of V. dahliae—producing much aerial and prostrate mycelium, heavy sporulation and many black microsclerotia. Approximately 10 μg/l of biotin had to be added to Dox's medium to stimulate maximum growth of this isolate, but it was unable to-translocate exogenously supplied biotin. The leakage of biotin by other fungi into culture media induced good growth. Evidence is presented to show preferential accumulation/synthesis of biotin in conidia. Many substances of reported biotin-sparing activity proved to be ineffective in replacing the biotin requirement of this isolate.

Verticillium Wilt of Hedysarum coronarium (‘Sulla’)

SUMMARYA hitherto unrecorded disease of Hedysarum coronarium L. caused by Verticillium dahliae Kleb. is described. The isolate had a marked requirement for biotin, a characteristic it retained even when reisolated from different types of inoculated host plants. Although pathogenicity was readily established in preliminary pot experiments, difficulties were encountered in later years in inducing infection. The most successful method of inoculation was by dipping the roots into a spore suspension of the isolate in water. Variations in a range of soil conditions appeared to have no effect upon the incidence of disease. The isolate induced wilt in garden pea, sweet pea, and sainfoin, caused dark‐brown discoloration of the vascular tissue of apparently healthy broad bean, runner bean, lupin and Brussels sprouts, but had no apparent effect upon lucerne, potato, antirrhinum, chrysanthemum and tomato.

The study of a strain of Candida utilis in the course of continuous cultivation on ethanol with special respect to biotin requirement.

It was found using a long-term continuous cultivation in a synthetic ethanol medium that a strain ofCandida utilis can permanently form a sufficient amount of growth factors. This fact is confirmed both by high yields of biomass and steady level of biotin in the yeast mass. Composition of proteins is the same as in yeasts grown on other carbon sources.

L-Glutamic Acid Fermentation

It is well known that biotin has a marked effect on L-glutamic acid fermentation. The authors have intended to find strains which are independent of the amounts of biotin in the culture medium. As a result, oleic acid-requiring mutants were obtained from a strain of Brevibacterium thiogenitalis which is an auxotroph for biotin. The growth of the mutant was remarkably stimulated by Tween 20, 40, 60, Ca ions and a small amount of corn steep liquor. And also, the mutant was found to have lost its requirement for biotin and showed growth response only to oleic acid or unsaturated fatty acids.

L-Glutamic Acid Fermentation

It is well known that biotin has a marked effect on l-glutamic acid fermentation.The authors have intended to find strains which are independent of the amounts of biotin in the culture medium. As a result, oleic acid-requiring mutants were obtained from a strain of Brevibacterium thiogenitalis which is an auxotroph for biotin. The growth of the mutant was remarkably stimulated by Tween 20, 40, 60, Ca ions and a small amount of corn steep liquor. And also, the mutant was found to have lost its requirement for biotin and showed growth response only to oleic acid or unsaturated fatty acids.The effect of biotin, oleic acid and other unsaturated fatty acids on the production of l-glutamic acid was investigated by using an oleic acid-requiring mutant of Brevibacterium thiogenitalis No. 653. The results described in the present paper showed that the oleic acid-requiring mutant D-248 produced a large amount of l-glutamic acid in the excess biotin-contaming media, and that oleic acid seemed to be completely replac...

A specific requirement for biotin in the synthesis of ornithine carbamoyltransferase by yeast.

1. Growth of a biotin-requiring strain of Saccharomyces cerevisiae in a medium containing a suboptimum concentration of biotin for growth caused a decreased synthesis of ornithine carbamoyltransferase as compared with yeast grown in a medium containing an optimum concentration of biotin. Inclusion of the biotin homologues norbiotin or homobiotin, but not bishomobiotin, in the biotin-deficient medium caused an appreciable increase in ornithine carbamoyltransferase synthesis without affecting growth or synthesis of total RNA and protein. The addition of norbiotin to biotin-deficient medium had no effect on the respiratory activity of the yeast or on the synthesis of aspartate carbamoyltransferase, acid phosphatase, beta-fructofuranosidase or malate dehydrogenase. 2. Synthesis of acetylornithine deacetylase and acetylornithine acetyltransferase was slightly diminished by the imposition of biotin deficiency, but the effect was not as great as on ornithine carbamoyltransferase synthesis. Incorporation of norbiotin in the biotin-deficient medium had no marked effect on the synthesis of any other arginine-pathway enzyme except ornithine carbamoyltransferase. 3. l-Ornithine induced synthesis of ornithine carbamoyltransferase in yeast grown in biotin-deficient medium, but in yeast grown in this medium supplemented with norbiotin it repressed synthesis of the enzyme. l-Arginine had no detectable effect on ornithine carbamoyltransferase synthesis by the yeast grown in biotin-deficient medium with or without norbiotin. l-Aspartate repressed synthesis of ornithine carbamoyltransferase in biotin-deficient yeast and completely nullified the stimulatory effect of norbiotin on synthesis of the enzyme in this yeast. 4. There was no increase in ornithine carbamoyltransferase synthesis in biotin-deficient yeast incubated in phosphate buffer, pH4.5, containing glucose and biotin or norbiotin. In biotin-deficient yeast suspended in complete medium containing an optimum concentration of biotin, there was an increase in ornithine carbamoyltransferase synthesis only after the onset of growth.

The comparative nutritional requirements of two sibling species Drosophila simulans and D. melanogaster

The quantitative nutritional requirements for larval growth, metamorphosis, and eclosion were determined in axenic culture for Drosophila simulans and D. melanogaster. The concentrations of essential nutrients allowing optimal larval development times were similar in the two species for the major nutrients and for pantothenic acid, folic acid, and choline, but differed for nicotinic acid, aneurin, pyridoxine, and especially riboflavin. No nutritional requirement for biotin in D. simulans could be demonstrated, and desthiobiotin had no antimetabolic effect at zero biotin levels.

Stimulation of Nitrobacter agilis by Biotin

Krulwich, Terry A. (Goucher College, Baltimore, Md.), and Helen B. Funk. Stimulation of Nitrobacter agilis by biotin. J. Bacteriol. 90:729-733. 1965.-Addition of biotin to nitrite-mineral medium greatly stimulated the autotrophic growth of four strains of Nitrobacter agilis. Comparisons of cultures of the organisms grown in parallel at 30 C in nitrite medium and in the medium supplemented with 150 mmug of biotin per ml showed that the vitamin promoted: (i) 2- to 4-fold greater rates of utilization of nitrite, and (ii) 100- to 1,000-fold greater populations of cells per milliliter. Avidin specifically inhibited the biotin stimulation of nitrite utilization at an avidin-biotin ratio of 133:1. Incubation of the four strains of N. agilis at 37 C imposed a requirement for biotin that could be met by daily addition of 150 mmug of the vitamin per ml of medium. The stimulatory effects of the vitamin at 30 C suggest that in N. agilis the synthesis of biotin is rate-limiting for growth.

VITAMIN REQUIREMENTS OF SEVERAL CELLULOLYTIC RUMEN BACTERIA.

Scott, H. W. (Ohio Agricultural Experiment Station, Wooster), and B. A. Dehority. Vitamin requirements of several cellulolytic rumen bacteria. J. Bacteriol. 89:1169-1175. 1965.-Four strains of cellulolytic bacteria recently isolated from in vitro rumen fermentations were used in this study. Nine water-soluble vitamins were tested in single-deletion and single-addition plus biotin experiments, each with and without charcoal-extracted casein hydrolysate. Bacteroides succinogenes A3C and B21a required only biotin under the above experimental conditions. Ruminococcus flavefaciens B34b showed an absolute requirement for biotin and was stimulated by p-aminobenzoic acid (PABA) in the single-deletion experiments. In the single-addition plus biotin experiments, PABA and, to a lesser extent, vitamin B(12) appeared to be required for maximal growth. The presence or absence of casein hydrolysate did not affect the vitamin requirements for the aforementioned three strains. In the single-deletion experiments, R. flavefaciens Cla showed an absolute requirement for biotin and, when casein hydrolysate was omitted, for B(12). When casein hydrolysate was present, no requirement for B(12) could be observed. In the single-addition experiments where the basal medium contained biotin and casein hydrolysate or B(12), PABA was required for maximal growth; however, the single deletion of PABA caused only slight retardation of growth. Investigation of the B(12) or casein hydrolysate requirement of Cla revealed that a mixture of purified amino acids simulating casein hydrolysate satisfied this requirement. Subsequent work indicated that this requirement could be satisfied by the amino acid methionine.

A Requirement for Biotin in Aspergillus niger when Grown on a Rhamnose Medium at High Temperature

A Requirement for Biotin in <i>Aspergillus niger</i> when Grown on a Rhamnose Medium at High Temperature

The taxonomic status of Lineola longa

Studies on the morphological and physiological characteristics of two strains of Lineola longa showed that these aerobic bacteria form endospores, are Gram-positive and have peritrichous flagella. On the basis of these observations it was concluded that L. longa is, in fact, a Bacillus species. This conclusion was strengthened by the fact that both strains exhibited nutritional characteristics often encountered among the aerobic sporeforming rods, such as requirements for biotin and thiamine in the growth medium. The base ratio of the DNA purified from L. longa was found to be within the range reported for a number of species of Bacillus. Additional investigation of the nutritional behavior of L. longa indicated that neither strain studied used glucose as an oxidizable substrate or as a carbon source. However, fatty acids and amino acids were readily utilized for growth. L. longa, on the basis of its morphological and physiological properties, could not be identified with any of the established species of Bacillus. It is suggested, therefore, that L. longa be recognized as a new species within the genus Bacillus and that it be renamed Bacillus macroides.

B-Vitamin Requirements of the Pink Bollworm1

The pink bollworm, Pectinophora gossypiella (Saunders), was aseptically reared to determine dietary B-vitamin requirements. Lack of any one of the following vitamins in a casein medium resulted in 100% larval mortality: Calcium pantothenate, folic acid, nicotinamide, pyridoxine, riboflavin, and thiamine. No larvae matured in the absence of biotin in amino acid medium. In a medium of wheat germ without added nicotinamide or calcium pantothenate, complete larval mortality occurred. Larval development with or without vitamin B12 in 3 different media was similar. Quantitative vitamin requirements on casein medium for calcium pantothenate, nicotinamide, riboflavin, thiamine, pyridoxine, and folic acid were 0.40, 0.20, 0.050, <0.050, 0.025, and 0.025mg/ 100g of medium, respectively. Quantitative biotin requirement on amino acid medium was 1 µg /100g of medium.

CAROTENOID PIGMENTS IN A MUTANT STRAIN OF VERTICILLIUM ALBO-ATRUM REINKE AND BERTHOLD.

THE only pigment known to be produced by wild-type strains of V. albo-atrum is of a dark melanoid type1. In an investigation involving the production of biochemical mutants in this fungus (using a strain originally isolated from wilted lucerne, 1962), an orange pigmented colony, which had a partial requirement for biotin, developed on plates of complete medium spread with conidia irradiated with ultra-violet at a dosage allowing for approximately 5 per cent survival. The irradiation was carried out using a sterile distilled water suspension of conidia in an open Petri dish at a distance of 20 cm from a 2537 A ‘Hanovia’ bactericidal tube model 11 (with ozone reducing filter). The close resemblance of this colony (designated M.5) in morphological details to wild-type led us to conclude that it is a mutant rather than a chance contaminant. The results of a preliminary extraction of M.5 suggested that several different carotenoids were present, and from further work, reported here, we have been able to identify some of these pigments.

ROLE OF MARINE FUNGI IN THE BIOCHEMISTRY OF THE OCEANS: III. GROWTH FACTOR REQUIREMENTS OF THE ASCOMYCETE HALOSPHAERIA MEDIOSETIGERA

Certain growth factor requirements of the cellulolytic marine ascomycete, Halosphaeria mediosetigera, in glucose – inorganic salts media, have been determined. Yeast extract, containing required nutrilites, proved superior to other empirical mixtures in the support of mycelial growth. Aqueous extracts of several varieties of wood flour could not replace yeast extract. Gulf Stream seawater was unable to provide the necessary growth factors. An absolute requirement for 5 to 10 μg thiamine per liter has been demonstrated for maximal mycelial development. Further data showed a requirement for biotin which was both partial and transient.