Abstract
Allantoinase (allantoin amidohydrolase, EC 3.5.2.5) catalyses the hydrolysis of allantoin to allantoic acid, a key reaction in the biosynthesis and degradation of ureides. This activity was determined in different tissues of French bean plants (Phaseolus vulgaris L.) which were grown under nitrogen-fixing conditions. Allantoinase activity was detected in all tissues analysed, but the highest levels of specific activity were found in developing fruits, from which allantoinase has been purified to electrophoretic homogeneity and further characterized. After diethylaminoethyl (DEAE)-Sephacel chromatography, two peaks showing allantoinase activity were obtained in the chromatographic profile and the corresponding proteins were independently purified. Total allantoinase activity was purified 200-fold, indicating the relevance of this enzymatic activity in French bean developing fruits, with allantoinase representing 0.5% of total soluble protein. Both proteins with allantoinase activity are monomeric with molecular masses of 45 and 42 kDa. The specific activities of the purified proteins were 560 and 295 units mg(-1), which correspond to turnover numbers of 25,200 and 12,100 min(-1), respectively. The two proteins have very similar biochemical properties showing Michaelis-Menten kinetics for allantoin with K(m) values of about 60 mM, with high optimal temperatures; are metalloenzymes; are inhibited by compounds reacting with sulphydryl groups; and are unaffected by reducing agents. All analysed tissues exhibited the two activities responsible for allantoin degradation, although one of them was the main form in leaves (the most photosynthetic tissue) and the other protein was the main form in roots (non-photosynthetic tissue). The allantoinase activity and distribution of both proteins have been analysed during fruit development. For both proteins, the allantoinase activity and distribution pattern were the same in plants growing either under nitrogen-fixing conditions or fertilized with nitrate.
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Claims 16
Claim · model-written summary
Allantoinase activity occurred in all analysed tissues and was highest in developing fruits.
- Taxon
- French bean
Phaseolus vulgarist0 - Organ / stage
- developing fruitse0
- Outcome
- Allantoinase activitye0
- Direction
- higheste0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Allantoinase activity was detected in all tissues analysed, but the highest levels of specific activity were found in developing fruits”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
Chromatography yielded two peaks of allantoinase activity, whose proteins were independently purified.
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- two peaks showing allantoinase activitye0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“After diethylaminoethyl (DEAE)-Sephacel chromatography, two peaks showing allantoinase activity were obtained in the chromatographic profile and the corresponding proteins were independently purified.”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
Total allantoinase activity was purified 200-fold.
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- Total allantoinase activitye0
- Value
- 200-folde0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Total allantoinase activity was purified 200-fold”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
Allantoinase represented 0.5% of total soluble protein in developing fruits.
- Taxon
- French beanPhaseolus vulgarise0
- Organ / stage
- developing fruitse0
- Gene
- allantoinasee0
- Outcome
- total soluble proteine0
- Value
- 0.5%e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“in French bean developing fruits, with allantoinase representing 0.5% of total soluble protein”
Claim · model-written summary
The first purified monomeric allantoinase protein had molecular mass 45 kDa.
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- molecular massese0
- Value
- 45e0
- Qualifier
- monomerice0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Both proteins with allantoinase activity are monomeric with molecular masses of 45 and 42 kDa.”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
The second purified monomeric allantoinase protein had molecular mass 42 kDa.
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- molecular massese0
- Value
- 42 kDae0
- Qualifier
- monomerice0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Both proteins with allantoinase activity are monomeric with molecular masses of 45 and 42 kDa.”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
The first purified protein had specific activity 560 units mg(-1).
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- specific activitiese0
- Value
- 560e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The specific activities of the purified proteins were 560 and 295 units mg(-1)”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
The second purified protein had specific activity 295 units mg(-1).
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- specific activitiese0
- Value
- 295 units mg(-1)e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The specific activities of the purified proteins were 560 and 295 units mg(-1)”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
The first protein had turnover number 25,200 min(-1).
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- turnover numberse0
- Value
- 25,200e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“turnover numbers of 25,200 and 12,100 min(-1), respectively”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
The second protein had turnover number 12,100 min(-1).
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- turnover numberse0
- Value
- 12,100 min(-1)e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“turnover numbers of 25,200 and 12,100 min(-1), respectively”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
Both proteins showed Michaelis-Menten kinetics for allantoin with Km about 60 mM and high optimal temperatures.
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- Michaelis-Menten kineticse0
- Value
- about 60 mMe0
- Qualifier
- with high optimal temperaturese0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The two proteins have very similar biochemical properties showing Michaelis-Menten kinetics for allantoin with K(m) values of about 60 mM, with high optimal temperatures”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
Both proteins were metalloenzymes.
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- metalloenzymese0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“are metalloenzymes; are inhibited by compounds reacting with sulphydryl groups; and are unaffected by reducing agents.”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
Compounds reacting with sulphydryl groups inhibited both proteins.
- Taxon
- French beanPhaseolus vulgarist0
- Treatment
- compounds reacting with sulphydryl groupse0
- Outcome
- allantoinase activitye1
- Direction
- inhibitede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“are inhibited by compounds reacting with sulphydryl groups”
t0“French bean (Phaseolus vulgaris)”
e1“Both proteins with allantoinase activity”
Claim · model-written summary
Reducing agents did not affect the proteins.
- Taxon
- French beanPhaseolus vulgarist0
- Treatment
- reducing agentse0
- Outcome
- allantoinase activitye1
- Direction
- unaffectede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“are unaffected by reducing agents”
t0“French bean (Phaseolus vulgaris)”
e1“Both proteins with allantoinase activity”
Claim · model-written summary
All analysed tissues contained both allantoin-degrading activities, with different predominant forms in leaves and roots.
- Taxon
- French beanPhaseolus vulgarist0
- Outcome
- two activities responsible for allantoin degradatione0
- Qualifier
- one of them was the main form in leaves (the most photosynthetic tissue) and the other protein was the main form in roots (non-photosynthetic tissue)e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“All analysed tissues exhibited the two activities responsible for allantoin degradation, although one of them was the main form in leaves (the most photosynthetic tissue) and the other protein was the main form in roots (non-photosynthetic tissue).”
t0“French bean (Phaseolus vulgaris)”
Claim · model-written summary
Both proteins had the same activity and distribution pattern under nitrogen fixation and nitrate fertilization.
- Taxon
- French beanPhaseolus vulgarist0
- Treatment
- fertilized with nitratee0
- Outcome
- allantoinase activity and distribution patterne0
- Direction
- the samee0
- Comparator
- nitrogen-fixing conditionse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“For both proteins, the allantoinase activity and distribution pattern were the same in plants growing either under nitrogen-fixing conditions or fertilized with nitrate.”
t0“French bean (Phaseolus vulgaris)”
The same questions in other papers
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2 papers in the same direction as this paper · 2 reporting no difference from an alternative
- positive
- positive
- negative
1 paper more in this group
See all 5 papers →nitrogen · physiology: enzyme activity · Phaseolus vulgaris (common bean)Positive and negative results
3 papers in the same direction as this paper · 3 reporting no difference from an alternative
- positive
- positive
- positive
3 papers more in this group
See all 7 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.