Isolation and partial characterization of a protease from <i>Cucurbita ficifolia</i>

Curotto, E.; González, Gustavo; O'Reilly, Sybil; Tapia, Guillermina

doi:10.1016/0014-5793(89)80162-0

Cited by 34 publications

(19 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…2), which agrees well with the value obtained by gel-filtration and indicates that the enzyme is monomeric. The molecular mass of the macluralisin polypeptide after deduction of carbohydrate (Curotto and Gonzalez 1989) Benincasa cerifera 50 (Kaneda and Tominago 1977) Cucumis melo 50 (Kaneda et al 1984) Trichosanthes cucumeroides A 50 (Lynn and Clevette-Radford 1988a) Helianthus annuus 25 (Rudenskaya et al 1987) components corresponded to 50 kDa, a value that agrees well with the data on cucumisin. Bacterial subtilisins, being substantially shorter, have molecular masses of about 29 kDa, while the sunflower leaf subtilisin has a molecular mass of 25 kDa.…”

Section: Resultssupporting

confidence: 80%

“…Nepenthes pitchers (Lobareva et al 1973) and a metalloproteinase from soybean leaves (McGeehan et al 1992), subtilisin-like enzymes from Helianthus (Rudenskaya et al 1987) and several cucurbitaceous plants (Kaneda and Tominago 1977;Kaneda et al 1984Kaneda et al , 1986Curotto and Gonzalez 1989;Uchikoba et al 1990) should be mentioned. The latter results are especially interesting as subtilisins used to be considered as a family of predominantly bacterial enzymes and only recently have representatives of this family been found in animals (Barr 1991).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Macluralisin ? a serine proteinase from fruits of Maclura pomifera (Raf.) Schneid.

Rudenskaya

Богданова

Revina

et al. 1995

Planta

View full text Add to dashboard Cite

A serine proteinase was isolated from fruits of Maclura pomifera (Raf.) Schneid. by affinity chromatography on bacitracin-containing sorbents and gel-filtration. The enzyme, named macluralisin, is a glycoprotein with a molecular mass of 65 kDa; its protein moiety corresponds to a molecular mass of 50 kDa. The substrate specificity of macluralisin towards synthetic peptides and insulin B-chain is similar to that of cucumisin, a subtilisin-like proteinase from melon fruit. The enzyme is completely inhibited by diisopropylfluorophosphate. Its amino-acid composition resembles that of a serine proteinase isolated from the Cucurbitaceae. The N-terminal sequence has 33% of its residues identical to those of the sequence of fungal subtilisin-like proteinase K. Hence, Maclura pomifera serine proteinase belongs to the subtilisin family, which seems to be broadly distributed in the plant kingdom.

show abstract

Section: Resultssupporting

confidence: 80%

Section: Introductionmentioning

confidence: 99%

Macluralisin ? a serine proteinase from fruits of Maclura pomifera (Raf.) Schneid.

Rudenskaya

Богданова

Revina

et al. 1995

Planta

View full text Add to dashboard Cite

show abstract

“…For this reason, they are extremely attractive to both the food and biopharmaceutical industries. 7,8 Antioxidant peptides are the most commonly occurring bioactive peptides in food. 3,4 Despite these effort, enzymatic hydrolysis conducted with the use of various enzymes of microbial, plant and animal origin, is still the main process for obtaining bioactive peptides from food products.…”

Section: Introductionmentioning

confidence: 99%

Antioxidant and antidiabetic activities of peptides isolated from a hydrolysate of an egg-yolk protein by-product prepared with a proteinase from Asian pumpkin (Cucurbita ficifolia)

et al. 2015

View full text Add to dashboard Cite

Bioactive peptides derived from food have been increasingly popular due to their therapeutic properties. Of particular importance are peptides with a multidirectional activity that can be used in the treatment and prevention of diet-related diseases. This paper attempts to utilize a by-product of phospholipid extraction from egg yolk as a source of peptides with a broad spectrum of biological activity. In addition, in this research we used a non-commercial enzyme obtained from Asian pumpkin, which has not been sufficiently researched in terms of its ability to release biopeptides from food proteins. In the present study the biological properties of peptides, derived from egg-yolk protein by-products (YP) remaining after phospholipid extraction, and their four synthetic analogs were investigated with regard to their antioxidant (radical scavenging capacity, Fe 2+ chelating effect, reducing power (FRAP)) and antidiabetic (a-glucosidase and DPP-IV inhibitory activities) properties. One of them, with the sequence LAPSLPGKPKPD, exhibited the highest antioxidant activity (free radical scavenging activity (6.03 mM Trolox eq per mg protein); FRAP (296.07 mg Fe 2+ per mg protein)). This peptide also revealed the strongest DPP-IV (361.5 mmol L À1 ) and a-glucosidase (1065.6 mmol L À1 ) inhibitory activities, a novel multifunctional effect for peptides from an egg-yolk hydrolysate.

show abstract

“…Like several other plant subtilisin-like endopeptidases, SEP-1 was most active at high temperatures and was relatively stable up to 50°C. Nevertheless, the optimum pH for its activity was lower than those of most similar enzymes, whose optimal values are around pH 9.0 (Kaneda and Tominaga 1977;Vera and Cornejero 1988;Curroto et al 1989;Rudenskaya et al 1998) or higher (Uchikoba et al 1990). Like that of SEP-1, the molecular weights of the cucumisin-like enzymes from Cucurbitaceae and from other plant families, are generally around 70 kDa (Vera and Cornejero 1988;Yamagata et al 1994;Ribeiro et al 1995;Rudenskaya et al 1995Rudenskaya et al , 1998Sabala et al 1997).…”

Section: Discussionmentioning

confidence: 95%

SEP-1 - a subtilisin-like serine endopeptidase from germinated seeds of Hordeum vulgare L. cv. Morex

Fontanini

Jones

2002

Planta

View full text Add to dashboard Cite

Proteolysis is crucial for all living cells. It regulates protein processing, intracellular protein levels and removes abnormal or damaged proteins from the cell, working as a cellular housekeeper. By means of proteolysis, cells can control the short-lived regulatory proteins that affect processes such as signal transduction and reception, transcription, division and cellular growth. Proteolysis also furnishes amino acids for the de novo synthesis of proteins. In germinating seeds, its main role is to degrade storage proteins into small peptides and amino acids that can be used by the embryo during autotrophic growth. We have isolated and purified a serine endopeptidase, one of the many proteolytic enzymes that occur in germinated barley seeds (green malt), using chromatofocusing and DEAE-, CM-, and size-exclusion chromatographies. The enzyme, named SEP-1, has a molecular weight of 70 kDa, as estimated by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography. SEP-1 was detected and measured by its ability to digest gelatin in gels and to hydrolyze the synthetic substrate N-succinyl Ala-Ala-Pro-Leu p-nitroanilide. The hydrolysis of the synthetic substrate was optimal at pH 6.5 and 50 degrees C with a K(m) of 2.6 mM. The enzyme was inhibited by phenylmethylsulfonyl fluoride and p-amidinophenyl methanesulfonyl fluoride but not by any other class-specific inhibitor, suggesting it was a serine endopeptidase. Its amino acid sequence was similar to those of other plant subtilisin-like serine peptidases (EC 3.4.21), especially to the cucumisin-like group. SEP-1 was present in resting seeds, and its activity increased during germination in all of the malted barley tissues except for the endosperm, where it never occurred, suggesting that the enzyme is not likely involved in storage-protein degradation.

show abstract

Isolation and partial characterization of a protease from Cucurbita ficifolia

Abstract: A protease from the pulp of Cucurbitaficifolia was purified. Its molecular mass was estimated to be about 60 kDa. Its maximum activity is in the alkaline region against azocollagen as substrate. The enzyme is inhibited by phenylmethylsulphonyl fluoride but not by EDTA and iodoacetic acid.

Cited by 34 publications

References 8 publications

Macluralisin ? a serine proteinase from fruits of Maclura pomifera (Raf.) Schneid.

Macluralisin ? a serine proteinase from fruits of Maclura pomifera (Raf.) Schneid.

Antioxidant and antidiabetic activities of peptides isolated from a hydrolysate of an egg-yolk protein by-product prepared with a proteinase from Asian pumpkin (Cucurbita ficifolia)

SEP-1 - a subtilisin-like serine endopeptidase from germinated seeds of Hordeum vulgare L. cv. Morex

Contact Info

Product

Resources

About