Optimization of conditions for the colorimetric determination of citrulline, using diacetyl monoxime

Boyde, T. R. C.; Rahmatullah, Mohammed

doi:10.1016/0003-2697(80)90404-2

Cited by 313 publications

(177 citation statements)

References 30 publications

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“…The reaction products, citrulline or carbamoylputrescine, were determined according to the method of Boyde & Rahmatullah (1980). A 250 ml aliquot of supernatant was mixed with 3 ml of a freshly prepared chromogenic solution containing 16.7 % sulphuric acid, 0.05 % butanedione monoxime and 0.01 % thiosemicarbazide.…”

Section: Methodsmentioning

confidence: 99%

Agmatine deiminase pathway genes in Lactobacillus brevis are linked to the tyrosine decarboxylation operon in a putative acid resistance locus

et al. 2007

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Agmatine deiminase pathway genes in Lactobacillus brevis are linked to the tyrosine decarboxylation operon in a putative acid resistance locus In lactic acid bacteria (LAB), amino acids and their derivatives may be converted into aminecontaining compounds designated biogenic amines, in pathways providing metabolic energy and/ or acid resistance to the bacteria. In a previous study, a pathway converting tyrosine to tyramine was detected in Lactobacillus brevis and a fragment of a gene possibly involved in the production of another biogenic amine, putrescine, from agmatine, was detected in the same locus. The present study was carried out to determine if Lb. brevis actually harbours two biogenic amineproducing pathways in the same locus and to investigate the occurrence of the two gene clusters in other bacteria. Sequencing of the DNA locus in Lb. brevis revealed a cluster of six genes that are related to previously reported genes of agmatine deiminase pathways but with marked differences such as two genes encoding putative agmatine deiminases rather than one. Heterologous expression of encoded enzymes confirmed the presence of at least one active agmatine deiminase and one amino acid transporter that efficiently exchanged agmatine and putrescine. It was concluded that the Lb. brevis gene cluster encodes a functional and highly specific agmatine deiminase pathway. Screening of a collection of 197 LAB disclosed the same genes in 36 strains from six different species, and almost all the positive bacteria also contained the tyrosine catabolic pathway genes in the same locus. These results support the hypothesis that the agmatine deiminase and tyrosine catabolic pathways belong to a genomic region that provides acid resistance and that is exchanged horizontally as a whole between LAB. INTRODUCTIONLactic acid bacteria (LAB) can produce metabolic energy or increase their acid resistance by using catabolic pathways that convert amino acids or their derivatives into aminecontaining compounds designated biogenic amines (Molenaar et al., 1993; Fernández & Zú ñiga, 2006). In bacteria, biogenic amines may serve physiological functions such as cell proliferation control, but they are more generally released into the medium as the end products of the catabolic pathways (Tabor & Tabor, 1985;Griswold et al., 2006). In fact, excretion of the amine is an essential step in the pathways (see below). LAB carrying amino acid catabolic pathways are an important source of biogenic amines in foods (Ten Brink et al., 1990;Stratton et al., 1991;Silla Santos, 1996). There is interest in avoiding the proliferation of these bacteria because ingestion of foods containing large quantities of biogenic amines may cause human health issues such as headache, palpitations, flushing or vomiting (Silla Santos, 1996). Recently the genes of diverse pathways producing biogenic amines were identified in LAB (for a review, see Fernández & Zú ñiga, 2006). Interestingly, the pathways are strain specific rather than species specific, suggesting that horizonta...

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Section: Methodsmentioning

confidence: 99%

Agmatine deiminase pathway genes in Lactobacillus brevis are linked to the tyrosine decarboxylation operon in a putative acid resistance locus

et al. 2007

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“…The L-citrulline assay was used to measure iNOS activity by a colorimetric assay of deproteinized samples, as previously described by Boyde et al 29 This method is based on the reaction between citrulline and diacetyl monoxime in the presence of sulfuric and phosphoric acids, which leads to a colored complex measured at 530 nm. L-citrulline was used as the standard and taken through the full assay procedure.…”

Section: Flow Cytometric Measurementsmentioning

confidence: 99%

Antitumor effects of a natural anthracycline analog (Aloin) involve altered activity of antioxidant enzymes in HeLaS3 cells

Nićiforović¹,

Adžić²,

Spasić³

et al. 2007

Cancer Biology & Therapy

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The antiproliferative and cytotoxic potential of the natural anthracycline aloin from Aloe vera was tested on human uterine carcinoma HeLaS3 cells. Aloin showed a pronounced antiproliferative effect at physiological concentration (IC 50 = 97 mM), caused cell cycle arrest in the S phase, and markedly increased HeLaS3 cell apoptosis (to 24%). In the concentration range of 20-100 mM, its action was accompanied by remarkable changes in the activity of almost all antioxidant enzymes: MnSOD activity was increased many fold, while CuZnSOD and iNOS activities were inhibited. Moreover, inhibition of CuZnSOD was shown to occur by direct aloin interaction with the enzyme. As catalase activity was not changed, it is suggested that such conditions were responsible for antiproliferative and cytotoxic effects owing to accumulation of H 2 O 2 . Aloin alone was a more potent proapoptotic agent than a 2 Gy fractional dose of ionizing radiation or a combination of the two. Compared to other currently used therapeutics, aloin, due to its less undesirable side effects and antimetastatic potential, may prove to be the agent of choice on which clinical protocols for the treatment of human cervical carcinoma should rely in future.

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“…The assay contained 1 mM L-arginine, 40 mM MES, pH 6.0, and 0.07 mg protein, in a final volume of 1.0 ml. After 1 h, the reaction was stopped by the addition of 0.075 ml 100 % (w/v) TCA, and the amount of citrulline formed was determined using diacetyl monoxime, as described by Boyde & Rahmatullah (1980). Catabolic ornithine carbamoyl transferase was determined by measuring 14 CO 2 release from L-[ 14 C-carbamoyl]citrulline.…”

Section: Methodsmentioning

confidence: 99%

Divergent polyamine metabolism in the Apicomplexa

et al. 2007

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The lead enzymes of polyamine biosynthesis, i.e. ornithine decarboxylase (ODC) and arginine decarboxylase (ADC), were not detected in Toxoplasma gondii [the limit of detection for ODC and ADC was 5 pmol min "1 (mg protein) "1 ], indicating that T. gondii lacks a forward-directed polyamine biosynthetic pathway, and is therefore a polyamine auxotroph. The biochemical results were supported by results obtained from data-mining the T. gondii genome. However, it was possible to demonstrate the presence of a highly active backconversion pathway that formed spermidine from spermine, and putrescine from spermidine, via the combined action of spermidine/ spermine N 1 -acetyltransferase (SSAT) or spermidine N 1 -acetyltransferase (SAT) and polyamine oxidase (PAO). With spermine as the substrate, T. gondii SSAT had a specific activity of 1.84 nmol min "1 (mg protein) "1 , and an apparent K m for spermine of 180 mM; with spermidine as the substrate, the SAT had a specific activity of 3.95 nmol min "1 (mg protein) "1 , and a K m for spermidine of 240 mM. T. gondii PAO had a specific activity of 10.6 nmol min "1 (mg protein) "1 , and a K m for acetylspermine of 36 mM. Furthermore, the results demonstrated that T. gondii SSAT was 50 % inhibited by 30 mM di(ethyl)norspermine. The parasite actively transported arginine and ornithine, which were converted via the arginine dihydrolase pathway to citrulline and carbamoyl phosphate, resulting in the formation of ATP via carbamate kinase. The lack of polyamine biosynthesis by T. gondii is contrasted with polyamine metabolism by other apicomplexans.

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Optimization of conditions for the colorimetric determination of citrulline, using diacetyl monoxime

Cited by 313 publications

References 30 publications

Agmatine deiminase pathway genes in Lactobacillus brevis are linked to the tyrosine decarboxylation operon in a putative acid resistance locus

Agmatine deiminase pathway genes in Lactobacillus brevis are linked to the tyrosine decarboxylation operon in a putative acid resistance locus

Antitumor effects of a natural anthracycline analog (Aloin) involve altered activity of antioxidant enzymes in HeLaS3 cells

Divergent polyamine metabolism in the Apicomplexa

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