Fractionation and purification of the thiol proteinases from papaya latex

Author(s):  
Philippe M. Dekeyser ◽  
Stefaan De Smedt ◽  
Joseph Demeester ◽  
Albert Lauwers
1979 ◽  
Vol 177 (2) ◽  
pp. 541-548 ◽  
Author(s):  
Baldev S. Baines ◽  
Keith Brocklehurst

A method of preparation of papain (EC 3.4.22.2) from relatively soluble types of latex of Carica papaya, including spray-dried latex produced by a controlled and relatively mild process, was devised. Spray-dried latex dissolves easily in water up to 350mg/ml at 22°C, which corresponds to approx. 230mg of protein/ml. When the usual method of preparation of crystalline papain contaminated only by its oxidation products, developed by Kimmel & Smith [J. Biol. Chem. (1954) 207, 515–531], is applied to spray-dried latex, the result is a preparation of papain heavily contaminated by chymopapains A and B (EC 3.4.22.6), and to a lesser extent by papaya peptidase A. This applies also to other types of papaya-latex currently commercially available, which, though less soluble than spray-dried latex, are more soluble than the types of latex available when the method of Kimmel & Smith (1954) was developed. This contamination is avoided by adjusting the concentration of the initial latex extract to 65mg of protein/ml (or less) before salt fractionation. For spray-dried latex this corresponds to 100mg of latex/ml. Papain isolated from spray-dried latex was characterized by using 2,2′-dipyridyl disulphide and 4-chloro-7-nitrobenzofurazan as thiol-specific reactivity probes and α-N-benzoyl-l-arginine ethyl ester as substrate. Papain isolated from this source appears to have the same catalytic-centre characteristics as papain isolated previously from latex produced by harsher methods. The catalysis of the hydrolysis of α-N-benzoyl-l-arginine ethyl ester by the mixture of thiol proteinases extracted from spray-dried latex by application of the method of Kimmel & Smith (1954) appears to obey Michaelis–Menten kinetics. The presence of the other enzymes results in an increase in the value of Km and a decrease in the catalytic-centre activity (kcat.) relative to the values for the catalysis by papain.


1993 ◽  
Vol 58 (10) ◽  
pp. 2321-2336 ◽  
Author(s):  
Zhong-wei Gu ◽  
John D. Spikes ◽  
Pavla Kopečková ◽  
Jindřich Kopeček

In cancer photodynamic therapy (PDT), improved efficiency of photosensitizer delivery to tumors may be obtained by binding them to targetable water soluble polymeric carriers. However, attachment of photosensitizers to Macromolecular carriers may alter their spectral and photosensitizing properties. In this study, a new monosubstituted phthalocyanine derivative, N-glycyl zinc(II) 4,9,16,23-tetraaminophthalocyanine (G-TAPC-Zn) was synthesized by the reaction of zinc(II) 4,9,16,23-tetraaminophthalocyanine (TAPC-Zn) with N-tert-butoxycarbonyl-glycine N'-hydroxybenzotriazole ester followed by deprotection of the tert-butoxycarbonyl (BOC) group. G-TAPC-Zn contains an aliphatic amino group suitable for attachment to water soluble polymeric carriers. By aminolysis of a polymeric precursor, an N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer containing oligopeptide (GFLG) side-chains terminated in p-nitrophenyl ester groups, with G-TAPC-Zn a polymeric derivative of the latter (P-GFLGG-TAPC-Zn) was synthesized. Spectral data indicated that in aqueous solutions P-GFLGG-TAPC-Zn formed aggregates. The degree of aggregation decreased with increasing concentration of detergents or organic solvents in buffer solutions. Consequently, the release of the drug from carrier catalyzed by thiol proteinases, papain or cathepsin B, took place only in the presence of detergents or organic solvents, i.e., under conditions with a lower probability of aggregate formation. Binding of G-TAPC-Zn to HPMA copolymers decreased the quantum yield of singlet oxygen generation from 0.24 to 0.063 and significantly increased its resistance to photobleaching.


2009 ◽  
Vol 40 (1) ◽  
pp. 122-124 ◽  
Author(s):  
Silas P. Rodrigues ◽  
Josemar S. Andrade ◽  
José A. Ventura ◽  
Patricia M. B. Fernandes
Keyword(s):  

2015 ◽  
Vol 2 (1) ◽  
pp. 41
Author(s):  
Umi Kulsum ◽  
Ratnawati Hendari ◽  
Siti Chumaeroh

Introduction: Research on the use of papaya and aloevera sap on wound healing each been done, but effects the combination of the two extracts on wound healing in diabetes mellitus condition has not been done.Purpose: This study aims to determine the effect of a gel combination of papaya latex and aloevera flesh extracts on the traumatics ulcer healing process in diabetic rats.Methods: True experimental laboratory research with the post-test-only control group design that performed on 28 male wistar rats which were divided intofour groups: DM and non-DM rat that treatment with povidone iodine and gel combination of papaya latex and aloevera flesh extract. The treatment was done for 7 days with smearing that was given 2x/day. Traumatics ulcer healing seen from the number of fibroblasts that then were analyzed by One WayAnova and post hoc LSD test.Results: The results showed the number of fibroblasts in DM and non-DM rat with povidone iodine were 44.6 and 42.73; in the DM and non-DM rat with a gel combination of papaya latex and aloe vera flesh extracts were 61.10 and 77.03. One Way Anova test resulted p-value = 0.000, it was mean that there were at least a significant difference on the number of fibroblasts in the two groups. Differences of the number fibroblasts was shown in all groups, except in the group DM and non-DM rats with povidone iodine (p = 0.764).Conclusion: Conclusion of the study was the provision of a gel combination of papaya latex and aloevera flesh extracts effect on the traumatics ulcer healing process in diabetic rats.


2020 ◽  
Vol 2 (1) ◽  
pp. 7
Author(s):  
Svetlana Olshannikova ◽  
Victoria Koroleva ◽  
Marina Holyavka ◽  
Alexander Pashkov ◽  
Valeriy Artyukhov

Plant enzymes such as ficin (EC 3.4.22.3), papain (EC 3.4.22.2) and bromelain (EC 3.4.22.4) are obtained from tropical plants. These biocatalysts belong to thiol proteases, in the active center of which cysteine is contained. Ficin, papain and bromelain have a wide substrate specificity, which provides a demand for their use in various industries. Enzymes in the free state are less commonly used; immobilized biocatalysts are the preferred form. The aim of this work was to determine the optimal concentration of a crosslinking agent in the covalent immobilization of ficin, papain and bromelain on a chitosan matrix. Ficin, papain and bromelain (Sigma) were chosen as objects of study. An acid-soluble chitosan (350 kDa, Bioprogress CJSC) was used as an immobilization carrier. The concentration range of glutaraldehyde (crosslinking agent) ranged from 1 to 25%. Suitable concentrations of glutaraldehyde for covalent immobilization were identified by the optimal ratio of protein content (mg per g of carrier), total activity (in units per ml of solution) and specific activity (in units per mg of protein). It was shown that for covalent immobilization of ficin and bromelain on a chitosan matrix, it is most promising to use 10% glutaraldehyde. For immobilization of papain on chitosan by covalent means, the concentration of glutaraldehyde equal to 20% is optimal.


Amino Acids ◽  
2009 ◽  
Vol 38 (1) ◽  
pp. 155-165 ◽  
Author(s):  
H. J. Cornell ◽  
W. Doherty ◽  
T. Stelmasiak
Keyword(s):  

2008 ◽  
Vol 49 (31) ◽  
pp. 4670-4673 ◽  
Author(s):  
Pierre Haquette ◽  
Barisa Talbi ◽  
Sigolène Canaguier ◽  
Samuel Dagorne ◽  
Céline Fosse ◽  
...  
Keyword(s):  
X Ray ◽  

2015 ◽  
Vol 90 (3) ◽  
pp. 284-293 ◽  
Author(s):  
F. Mansur ◽  
W. Luoga ◽  
D.J. Buttle ◽  
I.R. Duce ◽  
A. Lowe ◽  
...  

AbstractHymenolepis diminuta is a natural parasite of the common brown rat Rattus norvegicus, and provides a convenient model system for the assessment of the anthelmintic activity of novel drugs against cestodes. The experiments described in this paper indicate that treatment of rats infected with H. diminuta with a supernatant extract of papaya latex, containing a mixture of four cysteine proteinases, was moderately efficacious, resulting in a significant, but relatively small, reduction in worm burden and biomass. However, faecal egg output was not affected by treatment. In our experiments these effects were only partially dose-dependent, although specific inhibition by E-64 confirmed the role of cysteine proteinases as the active principles in papaya latex affecting worm growth but not statistically reducing worm burden. Data collected for a further 7 days after treatment indicated that the effects of papaya latex supernatant on worm loss and on worm growth were not enhanced. Our findings provide a starting point for further refinement in formulation and delivery, or assessment of alternative natural plant-derived cysteine proteinases in efforts to develop these naturally occurring enzymes into broad-spectrum anthelmintics, with efficacy against cestodes as well as nematodes.


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