FACTORS AFFECTING THE ACCUMULATION OF BIOTIN BY
            <i>LACTOBACILLUS ARABINOSUS</i>

Lichstein, Herman C.; Waller, James R.

doi:10.1128/jb.81.1.65-69.1961

Cited by 20 publications

(10 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Their experiments were performed by using microbiological assay for the determination of biotin uptake. As earlier work had indicated (16,17), the bacterial cells were found to accumulate biotin by active transport in the presence of exogenous glucose. In the absence of the carbohydrate, vitamin uptake resembled a carrier-mediated diffusion process.…”

supporting

confidence: 54%

Characterization of the Biotin Transport System in Saccharomyces cerevisiae

Rogers

Lichstein

1969

J Bacteriol

Self Cite

View full text Add to dashboard Cite

The characteristics of the biotin transport mechanism of Saccharomyces cerevisiae were investigated in nonproliferating cells. Microbiological and radioisotope assays were employed to measure biotin uptake. The vitamin existed intracellularly in both free and bound forms. Free biotin was extracted by boiling water. Chromatography of the free extract showed it to consist entirely of d-biotin. Cellular bound biotin was released by treating cells with 6 N H2SO4. The rate of biotin uptake was linear with time for 10 min, reaching a maximum at about 20 min followed by a gradual loss of accumulated free vitamin from the cells. Biotin was not degraded or converted to vitamers during uptake. Transport was temperature-and pH-dependent, optimum conditions for uptake being 30 C and pH 4.0. Glucose markedly stimulated biotin transport. In its presence, large intracellular free-biotin concentration gradients were established. Iodoacetate inhibited the glucose stimulation of biotin uptake. The rate of vitamin transport increased in a linear fashion with increasing cell mass. The transport system was saturated with increasing concentrations of the vitamin. The apparent Km for uptake was 3.23 X 10-7 M. Uptake of radioactive biotin was inhibited by unlabeled biotin and a number of analogues including homobiotin, desthiobiotin, oxybiotin, norbiotin, and biotin sulfone. Proline, hydroxyproline, and 7,8-diaminopelargonic acid did not inhibit uptake. Unlabeled biotin and desthiobiotin exchanged with accumulated intracellular "4C-biotin, whereas hydroxyproline did not. 1 Presented in part at the 68th Annual Meeting of the American Society for Microbiology, Detroit, Mich., 5-10 May 1968. Taken from a dissertation submitted by the senior author in partial fulfillment of the requirements for the Ph.D. degree in Microbiology. measuring exchange reactions and analogue inhibition patterns. In the experiments reported, vitamin uptake was measured in nonproliferating yeast cells. MATERIALS AND METHODS Chemicals. Crystalline d-biotin was purchased from the Sigma Chemical Co., St. Louis, Mo. Carbonyl-labeled 14C-biotin (57.5 mc/mmole) was obtained from Amersham-Searle Co., Des Plaines, Ill. The purity of the labeled biotin was checked by bioand radioautography. dl-Desthiobiotin was purchased from Nutritional Biochemicals Corp., Cleveland, Ohio. The biotin analogues d-homobiotin, dl-oxybiotin, and d-norbiotin were gifts from Hoffmann

show abstract

supporting

confidence: 54%

Characterization of the Biotin Transport System in Saccharomyces cerevisiae

Rogers

Lichstein

1969

J Bacteriol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Free biotin is defined as that amount of biotin, assayable with L. plantarum strain 17-5, which is released from bacterial cells upon short exposure to boiling water. Earlier studies on bound biotin formation (Lichstein and Ferguson, 1958;Lichstein and Waller, 1961) failed to reveal any cellular form of the vitamin other than bound biotin, since the boiling procedure used to stop the binding reaction released all free biotin from the cells. Data presented in Tables 1 and 2 show that free biotin was easily liberated from the bacterial cells.…”

Section: Resultsmentioning

confidence: 99%

“…Some properties of free and bound biotin. The extraction of free biotin from various bacterial cells and tissues has been described (Lichstein and Waller, 1961;Thompson, Eakin, and Williams, 1941). Free biotin is defined as that amount of biotin, assayable with L. plantarum strain 17-5, which is released from bacterial cells upon short exposure to boiling water.…”

Section: Resultsmentioning

confidence: 99%

“…Sanders and Leach (1964) characterized the uptake of lipoic acid by cells of Streptococcus faecalis(10 Cl) as an energy-requiring, temperature-dependent transport system which was easily saturated with lipoic acid and appeared to exist, at least in part, in the lipoprotein fraction of the cell membrane. The accumulation of bound biotin by cells of L. plantarum has been described (Lichstein and Ferguson, 1958;Lichstein and Waller, 1961).…”

mentioning

confidence: 99%

“…Microbiological assay procedure. The assay procedure was that of Lichstein and Waller (1961), except that sterilization was accomplished by autoclaving for 7 min at 121 C and the tubes were cooled immediately to reduce carmelization. Standard curves were prepared for each assay.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Biotin Transport and Accumulation by Cells of Lactobacillus plantarum I. General Properties of the System

Waller

Lichstein

1965

J Bacteriol

Self Cite

View full text Add to dashboard Cite

, AND HERMAN C. LICHSTEIN. Biotin transport and accumulation by cells of Lactobacillus plantarum. I. General properties of the system. J. Bacteriol. 90:843-852. 1965.-Resting cells of Lactobacillus plantarum were saturated with bound biotin by incubation in phosphate buffer with biotin and glucose for 2 hr. This bound biotin was stable to wide changes in temperature, pH, and reaction time. Free biotin could be eluted from the cells by suspending them in cold water or saline. Immersing the cells in boiling water for 30 sec released all free biotin. Recoveries of added biotin exceeded 92%. Free biotin uptake by bound biotin-saturated cells occurred by two mechanisms. One process was independent from temperature (Qlo, 1.25), pH, cellular metabolism, and inhibition by iodoacetate. The other mechanism was dependent upon temperature (Qlo, 2.58; optimum, 37 C), pH (optimum, 7.5), and active cellular metabolism, and was inhibited by iodoacetate. Activation energies of 3,700 and 13,800 cal per mole, respectively, were observed for glucose-independent and-dependent free biotin uptake. Both processes exhibited approximately the same degree of inhibition by homobiotin. Higher concentrations of homobiotin were required to inhibit growth than to inhibit free biotin uptake. Intracellular-extracellular ratios as high as 600 were established in the absence of glucose, whereas ratios of nearly 4,000 occurred in the presence of glucose. MATERIALS AND METHODS Organism and growth conditions. L. plantarum strain 17-5, which was employed in all studies, was maintained in stab culture on APT medium (Case Laboratories, Chicago, Ill.) by serial transfer every 2 to 4 weeks, incubated at 30 C overnight, and then refrigerated. Cells for experimental use were grown in the medium of Wright and Skeggs (1944) containing 5 X 10-4,ug of biotin per ml; the medium was modified by the substitution of cysteine for cystine, and the addition of folic acid (0.5 mg per liter) to improve growth. NaCl was omitted because the Difco Vitamin Free Casamoin Acids used as a source of amino acids contains 38% NaCl. Glucose was sterilized separately by auto-843

show abstract