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Ravinder K Grewal - One of the best experts on this subject based on the ideXlab platform.
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kinetics studies with Fruit Bromelain ananas comosus in the presence of cysteine and divalent ions
Journal of Food Science and Technology-mysore, 2015Co-Authors: Tajwinder Kaur, Amandeep Kaur, Ravinder K GrewalAbstract:The kinetics of cysteine and divalent ion modulation viz. Ca2+, Cu2+, Hg2+ of Fruit Bromelain (EC 3.4.22.33) have been investigated in the present study. Kinetic studies revealed that at pH 4.5, cysteine induced V-type activation of Bromelain catalyzed gelatin hydrolysis. At pH 3.5, Ca2+ inhibited the enzyme noncompetitively, whereas, both K-and V-type activations of Bromelain were observed in the presence of 0.5 mM Ca2+ at pH 4.5 and 7.5. Bromelain was inhibited competitively at 0.6 mM Cu2+ ions at pH 3.5, which changed to an uncompetitive inhibition at pH 4.5 and 7.5. An un-competitive inhibition of Bromelain catalyzed gelatin hydrolysis was observed in the presence of 0.6 mM Hg2+ at pH 3.5 and 4.5. These findings suggest that divalent ions modulation of Fruit Bromelain is pH dependent.
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kinetics studies with Fruit Bromelain ananas comosus in the presence of cysteine and divalent ions
Journal of Food Science and Technology-mysore, 2015Co-Authors: Tajwinder Kaur, Amandeep Kaur, Ravinder K GrewalAbstract:The kinetics of cysteine and divalent ion modulation viz. Ca2+, Cu2+, Hg2+ of Fruit Bromelain (EC 3.4.22.33) have been investigated in the present study. Kinetic studies revealed that at pH 4.5, cysteine induced V-type activation of Bromelain catalyzed gelatin hydrolysis. At pH 3.5, Ca2+ inhibited the enzyme noncompetitively, whereas, both K-and V-type activations of Bromelain were observed in the presence of 0.5 mM Ca2+ at pH 4.5 and 7.5. Bromelain was inhibited competitively at 0.6 mM Cu2+ ions at pH 3.5, which changed to an uncompetitive inhibition at pH 4.5 and 7.5. An un-competitive inhibition of Bromelain catalyzed gelatin hydrolysis was observed in the presence of 0.6 mM Hg2+ at pH 3.5 and 4.5. These findings suggest that divalent ions modulation of Fruit Bromelain is pH dependent.
Umesh H Hebbar - One of the best experts on this subject based on the ideXlab platform.
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efficacy of reverse micellar extracted Fruit Bromelain in meat tenderization
Journal of Food Science and Technology-mysore, 2014Co-Authors: Ram Saran Chaurasiya, P Z Sakhare, N Bhaskar, Umesh H HebbarAbstract:Reverse micellar extraction (RME) was used for the separation and purification of Bromelain from pineapple core and efficacy of RME purified Bromelain (RMEB) in tenderization of beef meat was compared with that of commercial stem Bromelain (CSB). RME resulted in reasonably high Bromelain activity recovery (85.0 %) and purification fold (4.0). Reduction in meat toughness was higher in RMEB treated meat (52.1 %) compared to raw (control) and CSB treated (26.7 %). Significant increase in water holding capacity (WHC) was observed in RMEB treated meat (91.1 %) as against CSB treated (55.6 %) and control (56.6 %). No change in cooking loss was observed in RMEB treated meat, whereas the loss increased by nearly 14.0 % in case of CSB treated. While the meat color was retained, trichloroacetic acid (TCA) soluble protein content increased due to hydrolysis of protein in RMEB treated meat. Scanning electron microscopy (SEM) analysis revealed that RMEB treatment completely ruptures myofibril tissues, indicating a higher degree of tenderization.
Aizi Nor Mazila Ramli - One of the best experts on this subject based on the ideXlab platform.
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gene expression analysis of Fruit Bromelain in ripening of ananas comosus cultivar md 2
Materials Science Forum, 2020Co-Authors: Wei Cheng Pang, Aizi Nor Mazila Ramli, Azzmer Azzar Abdul HamidAbstract:Fruit Bromelain is a proteolytic enzyme harbouring cysteine catalytic residue found abundantly in pineapple Fruit. The expression of cysteine proteases is usually regulated during Fruit ripening. In the present study, we aimed to study the expression and proteolytic activity level of Fruit Bromelain during the ripening stage of A. comosus cultivar MD 2. The gene expression of Fruit Bromelain was investigated via relative gene expression analysis using qPCR while the proteolytic activity of Fruit Bromelain was analysed via enzymatic assay using casein as a substrate. The qPCR analysis revealed that the expression of Fruit Bromelain was down-regulated 10-fold in ripe pineapple Fruits. Besides that, the unripe pineapple Fruits [1.9101 ± 0.0831 U/mL] had a higher proteolytic activity than the ripe MD 2 pineapple Fruits [1.1333 ± 0.0896 U/mL]. This result showed that the function of Fruit Bromelain may be related to the protection of young pineapple Fruits during the Fruit development stage.
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Gene Expression Analysis of Fruit Bromelain in Ripening of Ananas comosus Cultivar MD 2
'Trans Tech Publications Ltd.', 2020Co-Authors: Wei Cheng Pang, Aizi Nor Mazila Ramli, Azzmer Azzar Abdul HamidAbstract:Fruit Bromelain is a proteolytic enzyme harbouring cysteine catalytic residue found abundantly in pineapple Fruit. The expression of cysteine proteases is usually regulated during Fruit ripening. In the present study, we aimed to study the expression and proteolytic activity level of Fruit Bromelain during the ripening stage of A. comosus cultivar MD 2. The gene expression of Fruit Bromelain was investigated via relative gene expression analysis using qPCR while the proteolytic activity of Fruit Bromelain was analysed via enzymatic assay using casein as a substrate. The qPCR analysis revealed that the expression of Fruit Bromelain was down-regulated 10-fold in ripe pineapple Fruits. Besides that, the unripe pineapple Fruits [1.9101 ± 0.0831 U/mL] had a higher proteolytic activity than the ripe MD 2 pineapple Fruits [1.1333 ± 0.0896 U/mL]. This result showed that the function of Fruit Bromelain may be related to the protection of young pineapple Fruits during the Fruit development stag
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Comparative modelling studies of Fruit Bromelain using molecular dynamics simulation
'Springer Science and Business Media LLC', 2020Co-Authors: Pang, Wei Cheng, Aizi Nor Mazila Ramli, Azzmer Azzar Abdul HamidAbstract:Fruit Bromelain is a cysteine protease accumulated in pineapple Fruits. This proteolytic enzyme has received high demand for industrial and therapeutic applications. In this study, Fruit Bromelain sequences QIM61759, QIM61760 and QIM61761 were retrieved from the National Center for Biotechnology Information (NCBI) Genbank Database. The tertiary structure of Fruit Bromelain QIM61759, QIM61760 and QIM61761 was generated by using MODELLER. The result revealed that the local stereochemical quality of the generated models was improved by using multiple templates during modelling process. Moreover, by comparing with the available papain model, structural analysis provides an insight on how pro-peptide functions as a scaffold in Fruit Bromelain folding and contributing to inactivation of mature protein. The structural analysis also disclosed the similarities and differences between these models. Lastly, thermal stability of Fruit Bromelain was studied. Molecular dynamics simulation of Fruit Bromelain structures at several selected temperatures demonstrated how Fruit Bromelain responds to elevation of temperature
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Comparative structural analysis of Fruit and stem Bromelain from Ananas comosus
Food Chemistry, 2018Co-Authors: Aizi Nor Mazila Ramli, Nor Hasmaliana Abdul Manas, Hazrulrizawati Abd Hamid, Azzmer Azzar Abdul Hamid, Rosli Md. IlliasAbstract:Cysteine proteases in pineapple (Ananas comosus) plants are phytotherapeutical agents that demonstrate anti-edematous, anti-inflammatory, anti-thrombotic and fibrinolytic activities. Bromelain has been identified as an active component and as a major protease of A. comosus. Bromelain has gained wide acceptance and compliance as a phytotherapeutical drug. The proteolytic fraction of pineapple stem is termed stem Bromelain, while the one presents in the Fruit is known as Fruit Bromelain. The amino acid sequence and domain analysis of the Fruit and stem Bromelains demonstrated several differences and similarities of these cysteine protease family members. In addition, analysis of the modelled Fruit (BAA21848) and stem (CAA08861) Bromelains revealed the presence of unique properties of the predicted structures. Sequence analysis and structural prediction of stem and Fruit Bromelains of A. comosus along with the comparison of both structures provides a new insight on their distinct properties for industrial application.
Azzmer Azzar Abdul Hamid - One of the best experts on this subject based on the ideXlab platform.
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gene expression analysis of Fruit Bromelain in ripening of ananas comosus cultivar md 2
Materials Science Forum, 2020Co-Authors: Wei Cheng Pang, Aizi Nor Mazila Ramli, Azzmer Azzar Abdul HamidAbstract:Fruit Bromelain is a proteolytic enzyme harbouring cysteine catalytic residue found abundantly in pineapple Fruit. The expression of cysteine proteases is usually regulated during Fruit ripening. In the present study, we aimed to study the expression and proteolytic activity level of Fruit Bromelain during the ripening stage of A. comosus cultivar MD 2. The gene expression of Fruit Bromelain was investigated via relative gene expression analysis using qPCR while the proteolytic activity of Fruit Bromelain was analysed via enzymatic assay using casein as a substrate. The qPCR analysis revealed that the expression of Fruit Bromelain was down-regulated 10-fold in ripe pineapple Fruits. Besides that, the unripe pineapple Fruits [1.9101 ± 0.0831 U/mL] had a higher proteolytic activity than the ripe MD 2 pineapple Fruits [1.1333 ± 0.0896 U/mL]. This result showed that the function of Fruit Bromelain may be related to the protection of young pineapple Fruits during the Fruit development stage.
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Gene Expression Analysis of Fruit Bromelain in Ripening of Ananas comosus Cultivar MD 2
'Trans Tech Publications Ltd.', 2020Co-Authors: Wei Cheng Pang, Aizi Nor Mazila Ramli, Azzmer Azzar Abdul HamidAbstract:Fruit Bromelain is a proteolytic enzyme harbouring cysteine catalytic residue found abundantly in pineapple Fruit. The expression of cysteine proteases is usually regulated during Fruit ripening. In the present study, we aimed to study the expression and proteolytic activity level of Fruit Bromelain during the ripening stage of A. comosus cultivar MD 2. The gene expression of Fruit Bromelain was investigated via relative gene expression analysis using qPCR while the proteolytic activity of Fruit Bromelain was analysed via enzymatic assay using casein as a substrate. The qPCR analysis revealed that the expression of Fruit Bromelain was down-regulated 10-fold in ripe pineapple Fruits. Besides that, the unripe pineapple Fruits [1.9101 ± 0.0831 U/mL] had a higher proteolytic activity than the ripe MD 2 pineapple Fruits [1.1333 ± 0.0896 U/mL]. This result showed that the function of Fruit Bromelain may be related to the protection of young pineapple Fruits during the Fruit development stag
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Comparative modelling studies of Fruit Bromelain using molecular dynamics simulation
'Springer Science and Business Media LLC', 2020Co-Authors: Pang, Wei Cheng, Aizi Nor Mazila Ramli, Azzmer Azzar Abdul HamidAbstract:Fruit Bromelain is a cysteine protease accumulated in pineapple Fruits. This proteolytic enzyme has received high demand for industrial and therapeutic applications. In this study, Fruit Bromelain sequences QIM61759, QIM61760 and QIM61761 were retrieved from the National Center for Biotechnology Information (NCBI) Genbank Database. The tertiary structure of Fruit Bromelain QIM61759, QIM61760 and QIM61761 was generated by using MODELLER. The result revealed that the local stereochemical quality of the generated models was improved by using multiple templates during modelling process. Moreover, by comparing with the available papain model, structural analysis provides an insight on how pro-peptide functions as a scaffold in Fruit Bromelain folding and contributing to inactivation of mature protein. The structural analysis also disclosed the similarities and differences between these models. Lastly, thermal stability of Fruit Bromelain was studied. Molecular dynamics simulation of Fruit Bromelain structures at several selected temperatures demonstrated how Fruit Bromelain responds to elevation of temperature
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Comparative structural analysis of Fruit and stem Bromelain from Ananas comosus
Food Chemistry, 2018Co-Authors: Aizi Nor Mazila Ramli, Nor Hasmaliana Abdul Manas, Hazrulrizawati Abd Hamid, Azzmer Azzar Abdul Hamid, Rosli Md. IlliasAbstract:Cysteine proteases in pineapple (Ananas comosus) plants are phytotherapeutical agents that demonstrate anti-edematous, anti-inflammatory, anti-thrombotic and fibrinolytic activities. Bromelain has been identified as an active component and as a major protease of A. comosus. Bromelain has gained wide acceptance and compliance as a phytotherapeutical drug. The proteolytic fraction of pineapple stem is termed stem Bromelain, while the one presents in the Fruit is known as Fruit Bromelain. The amino acid sequence and domain analysis of the Fruit and stem Bromelains demonstrated several differences and similarities of these cysteine protease family members. In addition, analysis of the modelled Fruit (BAA21848) and stem (CAA08861) Bromelains revealed the presence of unique properties of the predicted structures. Sequence analysis and structural prediction of stem and Fruit Bromelains of A. comosus along with the comparison of both structures provides a new insight on their distinct properties for industrial application.
Debasish Bhattacharyya - One of the best experts on this subject based on the ideXlab platform.
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Fruit Bromelain derived peptides destabilize growth of amyloidal fibrils
bioRxiv, 2020Co-Authors: Sromona Das, Sangita Dutta, Ramesh Kumar Paidi, Subhas C Biswas, Umesh Chandra Halder, Debasish BhattacharyyaAbstract:{beta}-Amyloid deposition as fibrillar plaques in brain is the primary cause of Alzheimers disease. We report potency of cysteine protease Fruit Bromelain from pineapple in destabilising A{beta} fibrils. Bromelain peptide pool (Mw<500 Da) obtained mimicking human alimentary tract digestion inhibited fibrillation from monomeric and oligomeric states of A and irreversibly dissociated preformed fibrils into small oligomers of varied sizes. Time kinetics was followed by Thioflavin-T assay and microscopic imaging. Synthetic Bromelain peptides corresponding to A{beta} sticky region found using ClustalW analysis revealed specificity of peptides in destabilisation of amyloidal structures. Spectra of different molecular states of A{beta} obtained from application of 8-anilino-1-naphthalenesulfonic acid, circular dichroism and Fourier-Transformed Infrared spectroscopy collectively indicated interaction dependent structural change. Probable mechanism for fibril dissociation was thus predicted. Peptides relieved A{beta} cytotoxicity on pheochromcytoma cells and dissociated plaques in AD-type rats prepared by bilateral intracerebroventricular administration of A{beta} in rat brain cortex. Pineapple being a phytoceutical, its efficiency to disaggregate amyloid bodies warrant further investigation. GRAPHICAL ABSTRACT O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
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destabilization of human insulin fibrils by peptides of Fruit Bromelain derived from ananas comosus pineapple
Journal of Cellular Biochemistry, 2017Co-Authors: Sromona Das, Debasish BhattacharyyaAbstract:Deposition of insulin aggregates in human body leads to dysfunctioning of several organs. Effectiveness of Fruit Bromelain from pineapple in prevention of insulin aggregate was investigated. Proteolyses of Bromelain was done as par human digestive system and the pool of small peptides was separated from larger peptides and proteins. Under conditions of growth of insulin aggregates from its monomers, this pool of peptides restricted the reaction upto formation of oligomers of limited size. These peptides also destabilized preformed insulin aggregates to oligomers. These processes were followed fluorimetrically using Thioflavin T and 1-ANS, size-exclusion HPLC, dynamic light scattering, atomic force microscopy, and transmission electron microscopy. Sequences of insulin (A and B chains) and Bromelain were aligned using Clustal W software to predict most probable sites of interactions. Synthetic tripeptides corresponding to the hydrophobic interactive sites of Bromelain showed disaggregation of insulin suggesting specificity of interactions. The peptides GG and AAA serving as negative controls showed no potency in destabilization of aggregates. Disaggregation potency of the peptides was also observed when insulin was deposited on HepG2 liver cells where no formation of toxic oligomers occurred. Amyloidogenic des-octapeptide (B23-B30 of insulin) incapable of cell signaling showed cytotoxicity similar to insulin. This toxicity could be neutralized by Bromelain derived peptides. FT-IR and far-UV circular dichroism analysis indicated that disaggregated insulin had structure distinctly different from that of its hexameric (native) or monomeric states. Based on the stoichiometry of interaction and irreversibility of disaggregation, the mechanism/s of the peptides and insulin interactions has been proposed. J. Cell. Biochem. 118: 4881-4896, 2017. © 2017 Wiley Periodicals, Inc.
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enzymatic antimicrobial and toxicity studies of the aqueous extract of ananas comosus pineapple crown leaf
Journal of Ethnopharmacology, 2013Co-Authors: Sangita Dutta, Debasish BhattacharyyaAbstract:Abstract Ethnopharmacological relevance Various parts of the plant pineapple (Ananas comosus) are used in traditional medicine worldwide for treatment of a number of diseases and disorders. In folk medicine, pineapple leaf extract was used as an antimicrobial, vermicide, purgative, emmenagoogue, abortifacient, anti-oedema and anti-inflammatory agent. Compared to the Fruit and stem extracts of pineapple, information about its leaf extract is limited. The potential of pineapple crown leaf extract as an ethno-medicine has been evaluated in terms of its enzymatic activities related to wound healing, antimicrobial property and toxicity. Materials and methods Major protein components of the extract were revealed by 2-D gel electrophoresis followed by MS/MS analysis. Zymography, DQ-gelatin assay were performed to demonstrate proteolytic, fibrinolytic, gelatinase and collagenase activities. DNase and RNase activities were revealed from agarose gel electrophoresis. Antimicrobial activity was evaluated spectrophotometrically from growth inhibition. Sprague-Dawley rat model was used to measure acute and sub-acute toxicity of the extract by analyzing blood markers. Result The extract contains several proteins that were clustered under native condition. Proteomic studies indicated presence of Fruit Bromelain as major protein constituent of the extract. It showed nonspecific protease activity, gelatinolytic, collagenase, fibrinolytic, acid and alkaline phosphatase, peroxidase, DNase and RNase activities along with considerable anti-microbial property. The leaf extract did not induce any toxicity in rats after oral administration of acute and sub-acute doses. Conclusion Pineapple leaf extract is nontoxic, contains enzymes related to damage tissue repairing, wound healing and possibly prevents secondary infections from microbial organisms.
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preservation of natural stability of Fruit Bromelain from ananas comosus pineapple
Journal of Food Biochemistry, 2009Co-Authors: Rima Bhattacharya, Debasish BhattacharyyaAbstract:Pineapple extract, “Bromelain,” is a rich source of proteases having therapeutic benefits. The processed Fruit does not show proteolytic activity probably due to harsh conditions of sterilization. Storage of fresh Fruit at−4C without preservatives retained 75 ± 5% proteolytic activity after 180 days (with or without eight cycles of freeze–thawing in between) and was devoid of microbial contamination. Peroxidase, acid phosphatase and amylase activities were also retained by 75 ± 5%. Immunomodulatory activity of the fresh and the stored Fruit was comparable. For industrial scale, γ irradiation of 1–2 kGray is proposed to ensure sterility that does not affect enzyme activities. Under these conditions, no signs of freeze burning are observed. The ultraviolet-visible spectra of the fresh and stored Fruit extracts are also indistinguishable. The process is commercially viable and amply improves the quality of processed pineapple in terms of active “Bromelain.” PRACTICAL APPLICATIONS Bromelain, the Fruit extract of pineapple, is a rich source of beneficial enzymes primarily because of its medicinal properties. Because the Fruit grows in abundance in a short time, it requires preservation to prevent wastage. This work reports the absence of enzymatic activities in preserved pineapples commercially available in the global market because of harsh conditions of sterilization. Alternately, exposing to low dose of irradiation followed by preservation under frozen condition at −4C retains enzymatic and immunomodulatory activities of the Fruit. Being free from adverse effects of freeze burning, it offers a cost-effective way of proper preservation of pineapples retaining its medicinal properties. Promotion of pineapple in dietary consumption can prove to be beneficial to the society and businesses at large.
