Chromatography of 99 amino acids and other ninhydrin-reactive compounds in the Pickering lithium gradient system

Grunau, John A.; Swiader, John M.

doi:10.1016/0021-9673(92)80326-p

Cited by 62 publications

(19 citation statements)

References 8 publications

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“…The reaction was stopped after 15 or 30 min of incubation at 30°C by the addition of sulfosalicylic acid (final concentration, 3%). Samples of the supernatant obtained after centrifugation were analyzed by ion-exchange chromatography, followed by ninhydrin post-column derivatization (22). A PCX5200 postcolumn derivatizer (Pickering Laboratories, Mountain View, CA) was connected to a Waters 626 HPLC system (Waters, Milford, MA).…”

Section: Methodsmentioning

confidence: 99%

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Conversion of Methionine to Cysteine in Bacillus subtilis and Its Regulation

et al. 2007

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Bacillus subtilis can use methionine as the sole sulfur source, indicating an efficient conversion of methionine to cysteine. To characterize this pathway, the enzymatic activities of CysK, YrhA and YrhB purified in Escherichia coli were tested. Both CysK and YrhA have an O-acetylserine-thiol-lyase activity, but YrhA was 75-fold less active than CysK. An atypical cystathionine ␤-synthase activity using O-acetylserine and homocysteine as substrates was observed for YrhA but not for CysK. The YrhB protein had both cystathionine lyase and homocysteine ␥-lyase activities in vitro. Due to their activity, we propose that YrhA and YrhB should be renamed MccA and MccB for methionine-to-cysteine conversion. Mutants inactivated for cysK or yrhB grew similarly to the wild-type strain in the presence of methionine. In contrast, the growth of an ⌬yrhA mutant or a luxS mutant, inactivated for the S-ribosyl-homocysteinase step of the S-adenosylmethionine recycling pathway, was strongly reduced with methionine, whereas a ⌬yrhA ⌬cysK or cysE mutant did not grow at all under the same conditions. The yrhB and yrhA genes form an operon together with yrrT, mtnN, and yrhC. The expression of the yrrT operon was repressed in the presence of sulfate or cysteine. Both purified CysK and CymR, the global repressor of cysteine metabolism, were required to observe the formation of a protein-DNA complex with the yrrT promoter region in gel-shift experiments. The addition of O-acetyl-serine prevented the formation of this protein-DNA complex.Methionine plays a central role in a variety of cellular functions. This amino acid is the universal initiator of protein synthesis, and its derivative, S-adenosylmethionine (AdoMet), is involved in several cellular processes, including methylation and polyamine biosynthesis. Methionine synthesis and degradation are, therefore, tightly regulated.Bacillus subtilis, like several microorganisms (21, 28, 55), can use sulfate for the synthesis of organic sulfur metabolites, mostly cysteine, methionine, and AdoMet. As in enterobacteria (28), the sulfate assimilation pathway of B. subtilis involves uptake and activation of inorganic sulfate, followed by stepwise reduction to sulfide (Fig.

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Section: Methodsmentioning

confidence: 99%

“…A PCX5200 postcolumn derivatizer (Pickering Laboratories, Mountain View, CA) was connected to a Waters 626 HPLC system (Waters, Milford, MA). Amino acids were separated on a lithium ion-exchange column (3 mm [inner diameter] by 150 mm; Pickering) and detected at 570 and 440 nm on a Waters 2487 dual absorbance detector (22).…”

Section: Methodsmentioning

confidence: 99%

Conversion of Methionine to Cysteine in Bacillus subtilis and Its Regulation

et al. 2007

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“…Ionexchange chromatography followed by post column derivatization with ninhydrin was used (Spack man et al, 1958;Grunau and Swiader, 1992). Ninhydrin reacts with primary amino acids and hydrindantin to form Ruhemann`s Purple which is detectable at 570 nm.…”

Section: Post-column Derivatization and Chromatographic Conditionsmentioning

confidence: 99%

Determination of free amino acid content in the Slovenian dry-cured ham »Kraški pršut« and product characterization

Janeš¹,

Bolta²,

Škrlep³

et al. 2012

Acta Agriculturae Slovenica

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Determination of free amino acid content in the Slovenian drycured ham »Kraški pršut« and product characterizationThe aim of this study was to characterize the chemical profile of dry-cured ham selected and processed in accordance with the Slovenian consortium rules for »Kraški pršut« and to investigate the differences between the Biceps femoris (Bf) and the Semimembranosus (Sm) muscle. The free amino acid content, moisture, salt, protein, total nitrogen, non-protein nitrogen, proteolysis index, intramuscular fat and dry matter were analysed in 135 samples of dry-cured hams including both muscles. A modified method for sample preparation was developed for determination of free amino acids in dry-cured ham. The method was validated for linearity, limit of detection, limit of quantification, precision and uncertainty. According to the validation parameters the method is appropriate for the determination of free amino acid content in dry-cured ham. higher content for several free amino acids and total free amino acids were determined in the Bf muscle compared to the Sm muscle.Key words: meat products / dry-cured ham / Kraški pršut / chemical composition / free amino acids / methods / postcolumn derivatization / muscles / Biceps femoris / Semimembranosus Vsebnost prostih amino kislin v kraškem pršutu in opis izdelkanamen te študije je bil okarakterizirati kemijsko sestavo pršuta, ki je bil izbran in obdelan v skladu s slovenskimi pravili konzorcija za »Kraški pršut« in določiti razlike med Biceps femoris (Bf) in Semimembranosus (Sm) mišicama. Določili smo vsebnost prostih aminokislin, vlage, soli, beljakovin, skupnega dušika, ne-proteinskega dušika, proteolizni indeks, intramuskularne maščobe in suhe snovi v 135 vzorcih pršuta v obeh mišicah. modificirali smo pripravo vzorcev za določanje prostih aminokislin v pršut. metodo smo validirali in določili območje linearnosti, mejo detekcije, mejo kvantifikacije, ponovljivost in obnovljivost. Glede na dobljene rezultate je metoda primerna za določanje prostih aminokislin v pršutu. višje vsebnosti nekaterih prostih aminokislin in skupnih prostih aminokislin so bile v mišici Bf v primerjavi z mišico Sm.Ključne besede: mesni izdelki / pršut / Kraški pršut / kemijska sestava / proste aminokisline / metode / post-kolonska derivatizacija / mišice / Biceps femoris / Semimembranosus

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“…Samples were weighed, freeze dried and reweighed, and then extracted using 0.1 M hydrochloric acid. Amino acids were separated and quantified by the Poultry Science Laboratory at the University of Arkansas using the Pickering lithium gradient system with postcolumn ninhydrin derivatization and norLeu at 0.5 nmol mL 21 as an internal standard (Granau and Swiader, 1992). Amino acids that were analyzed included: Ser, Asn, Glu, Val, Ile, Leu, Phe, Asp, pseudoSer, Thr, Gln, Gly, Ala, citrulline, Cys, Met, Tyr, b-Ala, g-aminobutyric acid, Trp, ethanolamine, Lys, His, and Arg.…”

Section: Experiments 2: Ara and Leaf Ureide Response To Water Deficit mentioning

confidence: 99%

Inhibition of N2 Fixation in Soybean Is Associated with Elevated Ureides and Amino Acids

2005

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Decreased N 2 fixation in soybean (Glycine max) L. Merr. during water deficits has been associated with increases in ureides and free amino acids in plant tissues, indicating a potential feedback inhibition by these compounds in response to drought. We evaluated concentrations of ureides and amino acids in leaf and nodule tissue and the concurrent change in N 2 fixation in response to exogenous ureides and soil-water treatments for the cultivars Jackson and KS4895. Exogenous ureides applied to the soil and water-deficit treatments inhibited N 2 fixation by 85% to 90%. Mn fertilization increased the apparent catabolism of ureides in leaves and hastened the recovery of N 2 fixation following exogenous ureide application for both cultivars. Ureides and total free amino acids in leaves and nodules increased during water deficits and coincided with a decline in N 2 fixation for both cultivars. N 2 fixation recovered to 74% to 90% of control levels 2 d after rewatering drought-stressed plants, but leaf ureides and total nodule amino acids remained elevated in KS4895. Asparagine accounted for 82% of the increase in nodule amino acids relative to well-watered plants at 2 d after rewatering. These results indicate that leaf ureides and nodule asparagine do not feedback inhibit N 2 fixation. Compounds whose increase and decrease in concentration mirrored the decline and recovery of N 2 fixation included nodule ureides, nodule aspartate, and several amino acids in leaves, indicating that these are potential candidate molecules for feedback inhibition of N 2 fixation.Water-deficit stress is often a primary constraint of soybean (Glycine max) yields (Purcell and Specht, 2004). N 2 fixation in soybean is reportedly more sensitive to moderate water deficits than are many other plant processes including transpiration (Sall and Sinclair, 1991;Serraj and Sinclair, 1996) and leaf gas exchange (Durand et al., 1987;Djekoun and Planchon, 1991). It has been proposed that increased tolerance of N 2 fixation to water deficits would increase overall water-deficit tolerance in soybean.Despite much research effort in the area, the mechanisms influencing N 2 fixation response to limited soil water are not well understood. Accumulating evidence indicates that the decline in N 2 fixation during water deficits and genotypic differences in sensitivity to drought may be associated with levels of nitrogenous compounds, such as amino acids or ureides, in leaves or nodules of N 2 -fixing plants (Silsbury et al., 1986;Parsons et al., 1993;Oti-Boateng and Silsbury, 1993;Serraj et al., 1999a;Purcell et al., 2000;. How specific compounds regulate nodule activity is unclear, although several hypotheses have been proposed (Serraj et al., 1999b).The ureides, allantoin and allantoate, are the final products of N 2 fixation that are exported from soybean nodules to the shoot (McClure and Israel, 1979), where they are catabolized. While N 2 fixation declines in response to water deficits, shoot ureides often increase (de Silva et al., 1996; due to a decrease...

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Chromatography of 99 amino acids and other ninhydrin-reactive compounds in the Pickering lithium gradient system

Cited by 62 publications

References 8 publications

Conversion of Methionine to Cysteine in Bacillus subtilis and Its Regulation

Conversion of Methionine to Cysteine in Bacillus subtilis and Its Regulation

Determination of free amino acid content in the Slovenian dry-cured ham »Kraški pršut« and product characterization

Inhibition of N2 Fixation in Soybean Is Associated with Elevated Ureides and Amino Acids

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