Some studies on the effects of xanthble oxiase inhibitor a_opurIo 14-hydroxypyrazolo(34-)pyri.idel on alato met of soybean plants (Glychae max cv. T _ ) are reported.Soyben 'as asepticaly germinated for 96 hours on agar containing I _ilhlmlar allopurinol, contained only slight amounts of aUantoin, allantoic acid, and urea as compared with controls. Analysis of puries and pyrimidines of the allopurinol-treated seedlings sbowed marked accumulation of xanthine both in the cotyedons and seedlin axes. No hypoxanthine accumulation was found. Xanthine accumulation due to allopurinol treatment was relatively low after the cotyedons had fallen. For nodulated plants, allopurinol caused a sigificant drop in allantoin (+allantoic acid) in the stems and nodules, accompanied by a strking accumulatfon of xanthine in the nodules. The xanthine concentration in the nodules far exceeded that in the germinated slings. Allopurl at a concentration of 50 micromolar strongly inbibited xasthine oxidase prepared from soybean nodules.The results suggested that the main pathway of alantoin formation in soybean plants was through purine decomposition, via xanthine-uric acid. It was specially noted that a very active purine-decomposing system existed in soybean nodules.Allantoin is well known as one of the principal end products of nitrogen metabolism in animals and is produced by a biosynthetic sequence involving the formation of inosinic acid and its subsequent stepwise breakdown. Generally a variety of microorganisms, e.g. bacteria (2, 7), yeasts (30), fungi (3), and algae (1) have the ability to decompose oxypurines and consequently allantoin.Some higher plants such as maple, comfrey, and leguminous plants also produce and accumulate large amounts of allantoin, up to half the amount of their total N (24). In contrast to the animal systems which excrete allantoin as a waste material, these plants accumulate it within cells, transport it to various organs, and further reutilize its nitrogen (24,29). Several authors (5,10,17), by experiments using 'C-labeled purines or glycine, have demonstrated that allantoin or allantoic acid is formed through oxidative purine decomposition by a well defined reaction sequence in animals and microorganisms. Mothes (24) and Reinbothe (29) have, however, discussed in their reviews another plausible mode of allantoin synthesis, i.e. direct formation from allantoic acid via condensation of urea and glyoxylic acid. This simple reverse reaction may be supported by the finding that when ['4Clurea was fed to banana leaves allantoic acid was heavily labeled (12), and by a report of Brunel (9) suggesting an enzymic synthesis of allantoic acid from urea and glyoxylic acid in higher fungi. While it is likely that in higher plants most allantoin and allantoic acid are formed by the same pathway as that demonstrated in microorganisms and in animal tissues, the possibility that some might be made by the condensation of urea with glyoxylic acid has not been eliminated.Allopurinol [4-hydroxypyrazolo(3,4-d)pyrimidi...
For lindane degradation, a cell suspension of Clostridium rectum strain S-17 demands the addition of substrates such as leucine, alanine, pyruvate, a leucine-proline mixture, and molecular hydrogen. In the presence of leucine-proline mixture, lindane decomposed in parallel with isovaleric acid formation, and both lindane degradation and isovaleric acid formation were inhibited by monoiodoacetic acid, suggesting a close relation between lindane degradation and the Stickland reaction. Lindane was degraded by cell-free extracts of C. rectum in the presence of dithiothreitol (DTT). Radiogaschromatograms of n-hexane soluble metabolites from [14C] lindane showed the presence of monochlorobenzene and gamma-3,4,5,6-tetrachlorocyclohexene. Leucine, NADH, and NADPH were somewhat less active than DTT for lindane degradation in cell-free extracts. Reductive dechlorination seemed the major route of lindane degradation in cell-free extracts as well as in the intact cells of C. rectum.
Ultrasonic inactivation of Escherichia coli XL1-Blue has been investigated by high-intensity ultrasonic waves from horn type sonicator (27.5 kHz) utilizing the "squeeze-film effect". The amplitude of the vibration face contacting the sample solution was used as an indication of the ultrasonic power intensity. The inactivation of the E. coli cells by ultrasonic irradiation shows pseudo first-order behavior. The inactivation rate constant gradually increased with increasing amplitude of the vibration face and showed rapid increase above 3 microm (p-p). In contrast, the H2O2 formation was not observed below 3 microm (p-p), indicating that the ultrasonic shock wave might be more important than indirect effect of OH radicals formed by ultrasonic cavitation in this system. The optimal thickness of the squeeze film was determined as 2 mm for the E. coli inactivation. More than 99% of E. coli cells was inactivated within 180-s sonication at the amplitude of 3 microm (p-p) and 2 mm of the thickness of the squeeze film.
A BHC decomposer isolated from a paddy field soil of Shiga prefecture was identified to be Clostridium rectum. The most dominant degradation intermediate from T-BHC was identified to be ƒÁ-3,4,5,6-tetrachlorocyclohexene (ƒÁ-TCCH) by gas chromatography-mass spectrometry. Degradation of r-BHC took place rapidly in the early growth phase of C. rectum, but was remarkably reduced in the resting state, e.g., spore-formed cell or washed cell sus
To examine the influence of nodulation on the production of nitrogenous compounds, soybean plants (Glycine m a x var. Tamanishiki) were grown with or w i t h o u t N-fertilizer in the field, and the changes in amino-N and allantoin-N content in the different organs were determined throughout the growth period. I n the stem allantoin-N markedly increased after the flowering period and then decreased during seed formation. Allantoin accumulated in the pod (up to 70 per cent of total alcohol soluble-N) during pod formation, while in the seed the main N-compounds were amino acids, the allantoin coneentra~ tion being v e r y low. In well-nodulated soybeans grown w i t h o u t N-fertilizer, allantoin content in every plant organ was always high compared to beans grown with N-fertilizer, but amino-N content was comparatively low.Another experiment, in which soybean plants were allowed to form nodules by growing on a N-free medium, and in which a fixed-N supply was then controlled by the addition of various levels of ammonium, was made in a greenhouse. \ ¥ h e n nodules were formed, the subsequent addition of high concentrations of a m m o n i u m caused the accumulation of allantoin as well as the accumulation of amides and arginine. A possible role for allantoin and some aspects of its production in soybeans are discussed.
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