Purification and Characterization of β-<scp>d</scp>-Galactosidase and α-<scp>d</scp>-Mannosidase from Papaya (<i>Carica papaya</i>) Seeds

Ohtani, Kazuhiro; Misaki, Akira

doi:10.1080/00021369.1983.10865977

Cited by 4 publications

(6 citation statements)

References 6 publications

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“…During the course of the initial studies it was found that babaco exhibited considerable R-manosidase activity. It has been shown earlier that the latex of green fruits of papaya, a close relative to babaco and well-known for its high content of proteases (1,2), also contain significant levels of some glycosidases, notably β-glucosisdase, β-galactosidase, and R-mannosidase (3,4). Papaya is, to our knowledge, the only relative for which an R-mannosidase has been described.…”

Section: Introductionmentioning

confidence: 83%

Purification and Characterization of an α-Mannosidase from the Tropical Fruit Babaco (Vasconcellea × Heilbornii Cv. Babaco)

Blom

Reyes

Carlsson

2008

J. Agric. Food Chem.

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An alpha-mannosidase (EC 3.2.1.24) present in the lyophilized latex of babaco ( Vasconcellea heilbornii ) has been purified to apparent homogeneity by native PAGE. The purification involves a three-step procedure with successive anion exchange with Q Sepharose HP, lectin affinity chromatography using ConA Sepharose 4B, and gel filtration using Superdex 200 prep grade. The molecular mass was determined to be in the range of 260-280 kDa by Superdex 200 prep grade gel filtration, and isoelectric focusing showed a pI range between 5.85 and 6.55, suggesting different glycosylated isoforms. The optimal temperature for the alpha-mannosidase was determined to lie between 50 and 60 degrees C, and the optimal pH was 4.5 at 50 degrees C. The K(m) value for p-nitrophenyl alpha-mannopyranoside (pNPM) was found to be 1.25 mM and the V(max), 2.4 microkat mg(-1) at 50 degrees C and 1.94 microkat mg(-1) at 40 degrees C. The pure alpha-mannosidase was specific for mannose and did not display activity for any other tested synthetic substrates.

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Section: Introductionmentioning

confidence: 83%

Purification and Characterization of an α-Mannosidase from the Tropical Fruit Babaco (Vasconcellea × Heilbornii Cv. Babaco)

Blom

Reyes

Carlsson

2008

J. Agric. Food Chem.

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“…Moreover, Yashoda et al (2007) attributed the softening of ripened mango to the activities of endomannosidase and α-mannosidase. Ohtani and Misaki (1983) showed that the seeds of Carica papaya L. exhibited a high level of α-D-galactosidase and α-D-mannosidase activities. Other studies indicated that cellulase activities increase during the maturation of avocados, peaches, strawberries, tomatoes, and papayas (Ignacio et al 2011;Awad and Young 1979;Hobson 1981;Paull and Chen 1983).…”

Section: Introductionmentioning

confidence: 99%

The role of hydrolases in the loss of firmness and of the changes in sugar content during the post-harvest maturation of Carica papaya L. var solo 8

et al. 2012

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Fruit ripening is associated with many hydrolase activities involved in the softening of the fruit during the maturation. This study investigates the relationship between the loss of firmness along with the changes of sugar content and the enzymatic activities in Carica papaya L.var solo 8 during post-harvest storage. Three maturation stages (green immature: the fruit is entirely green, green mature: the fruit shows 1/32 yellow skin and fully mature: the fruit shows 1/8 yellow skin) have been selected and stored at 15, 22 and 28°C. The reduction of fruit firmness, total sugar contents, refractive index (% Brix) and enzymatic activities were measured. Low enzymatic activities (0.035 μmol/min/mg) were recorded in fruit harvested at the green immature stage with no significant (p≥0.05) effect on the softening while fruit harvested at the green mature and fully mature stages showed enzymatic activities 7 times as high as those of the green immature stage. These high enzymatic activities were responsible for the loss of firmness of the fruit. Accordingly, papayas at the green mature and fully mature stages displayed higher maxima of sugar content (4.8 g/100 g at 28°C at day 12, and 10.2 g/100 g at 22°C at day 8, respectively) at higher temperatures. Meanwhile in green immature papayas, the maximum was only 4.3 g/100 g at 22°C and day 12 of storage. The results show that the loss of firmness of the papaya was highly related to the hydrolytic enzyme activities and the sweet taste to the presence of simple sugars such as galactose liberated from the polysaccharide complexes.

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“…The optimum pH of the enzyme was around 5.0 with Man 5 GlcNAc 1 -PA as a substrate, suggesting that it is a kind of acidic glycosidases. The Ginkgomannosidase activity for Man 5 GlcNAc 1 -PA was significantly enhanced by Co 2þ but not by Zn 2þ or Ca 2þ , although it has been reported that Zn 2þ and Ca 2þ ions often activate some plant acidic -mannosidase [10][11][12][13][14] or some processing -mannosidase. 24,25) Furthermore, the enzyme activity was not inhibited by the addition of EDTA.…”

Section: Discussionmentioning

confidence: 94%

“…Concerning the plant -mannosidase, many acidicmannosidases have been purified and characterized from seeds and fruits. [9][10][11][12][13][14] From the viewpoint of the physiological function of -mannosidase in plant cells, these plant enzymes, which consist of two heterogenous subunits, have been considered to be involved in the degradation of N-glycosylated proteins in the vacuole or protein body, but not in the processing of N-glycans in the ER or Golgi apparatus. These plant -mannosidases require zinc or calcium ions for their activity and have optimum pH in the acidic region.…”

mentioning

confidence: 99%

Purification and Characterization of a Co(II)-Sensitive α-Mannosidase fromGinkgo bilobaSeeds

Woo

Miyazaki

Hara

et al. 2004

Bioscience, Biotechnology, and Biochemistry

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An alpha-mannosidase was purified from developing Ginkgo biloba seeds to apparently homogeneity. The molecular weight of the purified alpha-mannosidase was estimated to be 120 kDa by SDS-PAGE in the presence of 2-mercaptoethanol, and 340 kDa by gel filtration, indicating that Ginkgo alpha-mannosidase may function in oligomeric structures in the plant cell. The N-terminal amino acid sequence of the purified enzyme was Ala-Phe-Met-Lys-Tyr-X-Thr-Thr-Gly-Gly-Pro-Val-Ala-Gly-Lys-Ile-Asn-Val-His-Leu-. The alpha-mannosidase activity for Man(5)GlcNAc(1) was enhanced by the addition of Co(2+), but the addition of Zn(2+), Ca(2+), or EDTA did not show any significant effect. In the presence of cobalt ions, the hydrolysis rate for pyridylaminated Man(6)GlcNAc(1) was significantly faster than that for pyridylaminated Man(6)GlcNAc(2), suggesting the possibility that this enzyme is involved in the degradation of free N-glycans occurring in developing plant cells (Kimura, Y., and Matsuo, S., J. Biochem., 127, 1013-1019 (2000)). To our knowledge, this is the first report showing that plant cells contain an alpha-mannosidase, which is activated by Co(2+) and prefers the oligomannose type free N-glycans bearing only one GlcNAc residue as substrate.

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Purification and Characterization of β-d-Galactosidase and α-d-Mannosidase from Papaya (Carica papaya) Seeds

Cited by 4 publications

References 6 publications

Purification and Characterization of an α-Mannosidase from the Tropical Fruit Babaco (Vasconcellea × Heilbornii Cv. Babaco)

Purification and Characterization of an α-Mannosidase from the Tropical Fruit Babaco (Vasconcellea × Heilbornii Cv. Babaco)

The role of hydrolases in the loss of firmness and of the changes in sugar content during the post-harvest maturation of Carica papaya L. var solo 8

Purification and Characterization of a Co(II)-Sensitive α-Mannosidase fromGinkgo bilobaSeeds

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