Activities of Various Peptidases in Cotyledons of Germinating Peanut (<i>Arachis hypogaea</i>)

Mikola, Juhani

doi:10.1111/j.1399-3054.1976.tb04424.x

Cited by 19 publications

(5 citation statements)

References 9 publications

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“…Alkaline aminopeptidase activity has also been observed in other plants stich as peanuts (Mikola 1976) and Scots pine (Samia and Mikola 1975). However, this enzyme has been ptirified only from barley (Sopanen and Mikola 1975).…”

Section: Discussionmentioning

confidence: 86%

Purification and characterization of leucine aminopeptidase from kidney bean cotyledons

Mikkonen¹

1992

Physiol Plant

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Mikkonen, A. 1992. Purification and characterization of leucine aminopeptidase from kidney bean cotyledons. -Physiol. Plant. 84: 393-398.A leucine aminopeptidase (EC 3.4.11.1) was purified from cotyledons of resting kidney beans {Phaseolus vulgaris L. cv. Processor) by acidic extraction, ammonium sulfate fractionation and chromatography on DEAE-Sephacel, Sephacryl S-300, Mono Q HPLC and Superose HPLC columns. The yield of the 317-fold purified enzyme was 9%. On gel filtrations on Sephacryl S-300 and Superose HPLC the elution volumes of the enzyme corresponded to an M^ of 360 000. The enzyme gave one band on native gel electrophoresis and an eiectrophoretic titration in an immobilized pH gradient gave a single curve with a pi of 4.8. Two bands were observed in an SDS-gel electrophoresis with M, values of 58 000 and 60 000 both with and without reduction by 2-mercaptoethanol, indicating that subunits of the enzyme are not linked by disutphide bridges. The purified enzyme most rapidly liberated Leu and Ala ofthe N-termini of di-and oligopeptides, optimally at pH 9.0 ± 0.5. The etizvme was stable in the presence of glycerol, dithiothreitoi and Mg^^, while the latter also had an activating effect. Bestatin inhibited the enzyme competitively with Leu-Gly-Gly with a Kpvalue of 1.5 nM. These observations indicate that the purified aminopeptidase from the cotyledons of resting kidney beans corresponds to the cytosolic leucine aminopeptidase of mammalian tissues (EC 3.4.11.1). The high enzyme activity observed suggests that this aminopeptidase has an important roie in the production of free amino acids during germination.

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Section: Discussionmentioning

confidence: 86%

Purification and characterization of leucine aminopeptidase from kidney bean cotyledons

Mikkonen¹

1992

Physiol Plant

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“…This lack of sensitivity to DFP differentiates the present enzyme from the three carboxypeptidases of barley, all of which are completely inactivated by this reagent (13 3,11), and at least one gymnosperm (Scots pine; 19). Now, this activity has been purified about 3500-fold from germinated barley.…”

Section: Catalytc Propertiesmentioning

confidence: 79%

Purification and Partial Characterization of a Dipeptidase from Barley

Sopanen

1976

Plant Physiol.

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A peptidase hydrolyzing the dipeptide Ala-Gly optimally at pH 8 to 9 was pwified about 3500-fold from gerninated grains of baley (ffordeum vulgare L.). According to disc electrophoresis in the presence of sodium dodecyl sulfate, the preparation was about 90% pure.The enzyme preparation hydrolyzed all of the 15 neutnl dipeptides tested, but showed no activity on Lys-Gly or Asp-Ala. Three tripeptides and four synthetic aminopeptidase substrates were hydrolyzed at less than 0.1% of the rate for Ala-Gly; inhibition and inactivation tests indicated that these reactions were due to some contaminating aminopeptidase(s). The results suggest that the purified enzyme is a specific dipeptidase. Km values were determined for six dipeptides at pH 8.8; they varied from 0.3 to 15.8 mm. The enzyme was strongly inhibited both by metal chelators and by sulfhydryl reagents. The elution volumes of the enzyme in gel chromatography on Sephadex G-150 and Ultrogel AcA 22 corresponded to molecular weight 130,000 and 175,000, respectively. The migration rate in sodium dodecyl sulfate disc electropboresis indicated a subunit molecular weight of 50,000.The barley enzyme is remarkably similar to severl mmmalan and microbial dipeptidases.In 1929, Linderstrom-Lang and Sato (9) demonstrated that germinated barley contains high activities of two peptidases, one hydrolyzing Leu-Gly-Gly and Leu-Gly at pH 8.6, the other acting on Ala-Gly at pH 7.8. In a previous paper, we reported the purification and characterization of the former enzyme (22). This barley peptidase appeared remarkably similar to the mammalian and bacterial leucine aminopeptidases both in chemical and in enzymic properties.This paper contains a corresponding report for the Ala-Glysplitting barley peptidase. The results indicate that the enzyme is a specific dipeptidase, which is clearly similar to several mammalian and microbial dipeptidases.MATERIALS AND METHODS Plant Material. Pirkka "high enzyme" malt was obtained from Lahden Polttimo Oy, Lahti, Finland. Pirkka is a Finnish 6-row cultivar of barley (Hordeum vulgare L.).Reagents. The substrates were obtained from the following sources:

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“…It was reported in Arachis hypogea L. cv. Virginia 56-R seeds (Mikola, 1976) that CP has not an important role on protein mobilization.…”

Section: Discussionmentioning

confidence: 94%

Protein Mobilization and Proteolytic Enzyme Activities during Seed Germination of Broad Bean (Vicia faba L.)

Kırmızı

Guleryuz

2006

Zeitschrift Für Naturforschung C

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The protein mobilization from attached and detached seeds of Vicia faba L. cv. Eresen 87 (Fabaceae) was investigated. While the total soluble protein content decreased, the free amino acid content increased during the 7 days germination period. Among the three proteolytic enzymes, only endopeptidase activity was found to be affected by the removal of the embryonic axis. Leucine aminopeptidase activity was high at the beginning, then it decreased; carboxypeptidase activity reached the highest value at day 5. In order to examine the effects of plant growth regulators on detached cotyledons incubated with plant growth regulators [10 Ð4 m benzyladenine (BA), gibberellic acid (GA 3 ), indole acetic acid (IAA) and 10 Ð5 m abscisic acid (ABA)], only benzyladenine was found promotive on protein mobilization. Our results suggest that the removal of the embryonic axis in seeds of Vicia faba L. cv. Eresen 87 decreases protein mobilization and endopeptidase activity.

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Activities of Various Peptidases in Cotyledons of Germinating Peanut (Arachis hypogaea)

Cited by 19 publications

References 9 publications

Purification and characterization of leucine aminopeptidase from kidney bean cotyledons

Purification and characterization of leucine aminopeptidase from kidney bean cotyledons

Purification and Partial Characterization of a Dipeptidase from Barley

Protein Mobilization and Proteolytic Enzyme Activities during Seed Germination of Broad Bean (Vicia faba L.)

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