Extraction Of Bromelain Research Articles

In vitro antiplatelet activity tests conducted on bromelain extracted from the pineapple plant and purified by ion-exchange chromatography using a diethylaminoethyl cellulose xolumn

The present study is aimed at isolating and purifying the bromelain enzyme extracted from the cores of the fruit of the pineapple plant (Ananas comusus). The process of purification of the crude enzyme extract included, in sequence, fractionation through ammonium sulfate precipitation, dialysis, and ion-exchange chromatography. Fractionation of the crude enzyme extract with ammonium sulfate resulted in an increase of the specific activity. The fraction with the highest specific activity was that obtained by precipitation with 50–80 % ammonium sulfate. For this fraction, the specific activity was 8.683 U/mg and the degree of purity of the enzyme was 160 times greater than that of the crude enzyme extract. Further purification by column chromatography using diethylaminoethyl cellulose (DEAE-C) as ion-exchange medium produced six fractions (out of 92) characterized by peaks in proteolytic activity. In particular, the highest proteolytic activity (10.530 U/mg) was displayed by the fifth such fraction. The platelet aggregation inhibitory activity of the DEAE-C-purified bromelian extract was assayed through the Born’s method that uses platelet-rich plasma and asetosal as positive control. The aggregation percent of this bromelain extract was measured to be 49.70% and its inhibition percent 46.89 %.

Recent Developments of Reverse Micellar Techniques for Lysozyme, Bovine Serum Albumin, and Bromelain Extraction.

Biomolecules produced by living organisms can perform vast array of functions and play an important role in the cell. Important biomolecules such as lysozyme, bovine serum albumin (BSA), and bromelain are often studied by researchers due to their beneficial properties. The application of reverse micelles is an effective tool for protein separation from their sources due to the special system structure. Mechanisms of transferring biomolecules and factors that influence the extraction of biomolecules are reviewed in this paper. The enhancement of biomolecule extraction could be achieved depending on the properties of reverse micelles. This paper provides an overall review on lysozyme, BSA, and bromelain extraction by reverse micelle for various applications.

Sensory acceptance and shelf life of fresh cheese made with dry bromelain extract as a coagulating agent

The objective of this research was to evaluate the sensory acceptance and shelf life of a fresh cheese using dry bromelain extract as a coagulating agent. The dry bromelain extract was used in three concentrations (5%, 10% and 15% w / w) and the milk was used craw and pasteurized, for a total of six treatments under study. A sensory analysis was carried out and the useful life (sensorial, physicochemical and microbiological) of the treatment with greater acceptance was determined. The results showed that the cheeses obtained with greater sensory acceptance were those made with 5% w / w of bromelain dry extract regardless of the type of milk used (raw or pasteurized). The useful life of the selected product (T2) at the physicochemical level is 26 days, however sensory and microbiologically the product is of good quality until day 14. Therefore, the use of dry bromelain extract as a vegetable coagulant showed a great potential as a substitute for traditional rennet.

Antibacterial activity of hydrolysate protein from Etawa goat milk hydrolysed by crude extract bromelain

A study has been conducted on the potential of hydrolysate protein from Etawa goat milk protein which is hydrolysed by bromelain crude extract enzyme. The bromelain enzyme is extracted from the leaf of honey pineapple fruit. The research steps are: bromelain extraction, bromelain characterization, preparation of goat milk protein, hydrolysis of goat milk protein with bromelain and antibacterial activity test of protein hydrolysate fractions against Escherichia coli and Staphylococcus aureus bacteria by diffusion method. The bromelain crude extract enzyme has the following characteristics: optimum pH is 6, optimum temperature is 55 °C, activity is inhibited by EDTA metal chelating compound and Cu2+ and Zn2+ metal ions, whereas Ca2+ and Mg2+ metal ions increase activity. Inhibition of activity by EDTA shows that the bromelain enzyme is a metalloenzyme. Ca2+ and Mg2+ metal ions act as activators, while Cu2+ and Zn2+ metal ions are inhibitors. Protein preparation of goat milk produces two types of protein namely casein and whey. The result of hydrolysis of both protein types with characterized bromelain enzymes are hydrolysate proteins, which are then tested for their antibacterial activity. The results showed that the hydrolysate protein was able to inhibit the growth of Escherichia coli and Staphylococcus aureus bacteria.

Properties of bromelain extract from different parts of the pineapple variety Morris

Abstract The purpose of this study is to characterize the bromelain extract from pineapples which belong to the Morris variety. The bromelain was extracted from two different batches of Morris pineapples; one with crown and the other without crown. The weight of each part such as flesh, peel, core, stem and crown were 45–52%, 28–29%, 13–14%, 5–6% and 7% (w/w), respectively. The physiochemical properties of each part of the pineapple were determined. Results showed that the pH, TSS (oBrix), volume of extract (ml), total protein (mg/ml) and bromelain activity (CDU/ml) of the pineapple were 3–4, 1–6 oBrix, 850–6000 ml, 21–98 mg/ml and 95–251 CDU/ml, respectively. Both species showed that the peel and flesh gave the highest bromelain activity and protein contents, whereas the extract from the stem revealed the lowest values. The storage stability study showed that bromelain activities of the extract decreased with time under refrigerated condition (5–8 °C).

Optimization of bromelain isolation from pineapple byproducts by polysaccharide complex formation

A simple method for bromelain extraction from industrial pineapple residues (stems and peels) was developed and optimized through factorial experimental design. The developed methodology, based on precipitation with carrageenan, represents an alternative to the use of organic solvents and inorganic salts (common industrial precipitation) and allows achieving extracts with high bromelain purity.High recovery yield – 80–90% - of active bromelain was obtained for both crude juices (stems and peels) making possible to obtain ca. 0.3 g of bromelain from 100 g of pineapple byproducts using a low polysaccharide concentration (0.2–0.3% w/v).

Specific Activity of Pineapples Bromelain Extracts at Different Purification Stages Obtained from Different Agro-Ecological Zones of Thika Region, Kenya

Specific Activity of Pineapples Bromelain Extracts at Different Purification Stages Obtained from Different Agro-Ecological Zones of Thika Region, Kenya

Enzymatic activity of bromelain isolated pineapple (Ananas comosus) hump and its antibacterial effect on Enterococcus faecalis

Background: Enterococcus faecalis is the frequent cause of oral infections, such as periodontitis, infected root canals, and peri-radicular abscesses. Pineapple ( Ananas comosus ) fruit contains bromelain, one of proteolytic enzymes associated with several health benefits. Bromelain has been shown to promote healthy digestion, stimulate the immune system, improve cardiovascular conditions, and accelerate wound healing. Bromelain compounds possess anti-inflammatory and anticancer properties and exhibit antibacterial activity. Objective: To analyze the enzymatic activity of bromelain extracted from pineapple hump and investigate the antibacterial effect of bromelain against E. faecalis. Methods: Pineapple hump was dried and extracted with maceration technique. Further purification was obtained by ammonium sulfate fractionation, dialysis and ion exchange chromatography. Minimum inhibitory concentration (MIC) tests using diffusion and dilution techniques tested the antibacterial activity of the bromelain extract on E. faecalis. A one-way analysis of variance (ANOVA) test analyzed the significance of the differences in the E. faecalis inhibition zones after treatment with a range of bromelain extract concentrations. Differences were considered statistically significant if p < 0.05. Results: The specific activity of bromelain in the crude extract was 62.89 U/mg. Furthermore, bromelain activity using ammonium sulfate fractionation was 50.99 U/mg, dialysis was 54.59 U/mg, and ion exchange chromatography was 152.38 U/mg. The bromelain extract showed effective inhibitory and bactericidal activity against E. faecalis. The results of the inhibition test using a bromelain extract purified by ion exchange chromatography demonstrated that a concentration as small as 12.5% was effective in inhibiting the growth of E. faecalis (p < 0.05). Conclusion: The highest enzymatic activity of bromelain was found after purification with ion exchange chromatography. Bromelain exerted an antibacterial effect against a potent endodontic pathogen, but further studies are needed to explore this effect.

Production of Plutella xylostella bioinsecticide for brassicaceae plant based on bromelain enzyme extracted from pineapple peel and isothiocyanate extracted from broccoli stem and radish peel

Brassicaceae plant is a commodity of edible plant that is very important for Indonesia, with cabbage worm (Plutella xylostella) as its main pest. Pest control with biopesticide is needed to preserve environment and increase crops production. Several biological substance is toxic to cabbage worm, including bromelain enzyme and isothiocyanates. Extraction of bromelain is experimented by varying mixing duration (15, 30, 45, 60 mins) and solvent type (distilled water and phosphate buffer). Extraction of isothiocyanate is experimented by varying the solvent ratio (1:1, 1:2, 1:3), using dichloromethane as solvent. For the broccoli stem sample, phosphate buffer is added to maintain pH. Enzyme activity test is used with product tyrosin concentration as standard. Isothiocyanate is determined with GC-MS analysis. The most effective extraction method for bromelain is with distilled water as solvent with 15 min mixing time. The most effective extraction method for isothiocyanate is with 1:1 solvent ratio at 37°C temperature without adding phosphate buffer. Efficacy of each sample to cabbage worm is also experimented by feeding it broccoli leaf spread with sample and observing the mortality rate. Bromelain and isothiocyanate proved to be toxic to cabbage worm and can be used as alternative bioinsecticide with the highest mortality of cabbage worm is by isothiocyanate from radish peel, reaching 100%.

A simple method for purification of bromelain in a thermosensitive triblock copolymer-based protection system and recycling of phase components

ABSTRACTIn the work, we chose stem bromelain as a model to investigate the storage and purification of bromelain from pineapple peel. Extraction of bromelain from pineapple peel using a two-stage aqueous two-phase extraction system composed of a thermoseparating copolymer EOPOEO and K2HPO4. Bromelain predominantly partitioned to the EOPOEO-rich phase and then re-extract to the top dilute phase. The recovery of enzyme activity (68.6%) and purification factor (6.53) were determined under optimum conditions. The EOPOEO-rich phase and salt were recycled, and the recovery of enzyme activity could reach up to 60%. This method has been proved to obtain highly purified and stable bromelain.

An integrated approach for pineapple waste valorisation. Bioethanol production and bromelain extraction from pineapple residues

Waste management is critical for the food industry for which there is increasing interest in food waste valorisation processes. Pineapple waste, an abundant agro-industrial residue, is studied as a low-cost material for the generation of different value-added products. The work succeeds in obtaining bioethanol and proteolytic enzymes from these residues and, accordingly, integrated approaches for pineapple waste valorisation combining the production of bioethanol and bromelain in a unique process are suggested. There are several potential uses for the products obtained, while bioethanol is a well-known alternative to petroleum-based transport fuels, bromelain is mainly used in the food and pharmaceutical industries, and it has also applications in the cosmetics, textile, leather and detergents ones. Proposals are based on the optimization of bioethanol production through different fermentation and saccharification processes: direct fermentation of the liquor, consecutive saccharification and fermentation of the solid waste and simultaneous saccharification and fermentation of the solid waste. Simultaneous saccharification and fermentation increased ethanol production (5.4 ± 0.1% v/v) as compared to direct fermentation (4.7 ± 0.3%) and saccharification and fermentation of the solid waste (4.9 ± 0.4% v/v) processes. On the other hand, bromelain separation was accomplished using membrane separation techniques (microfiltration and ultrafiltration), and further stabilization of the concentrated stream by freeze-drying. An increased protein concentration after downstream processes was confirmed by the Lowry analytical method (11.5 ± 1.2 to 21.0 ± 1.3 mg/mL in the retentate), and the proteolytic activity of the lyophilized powder was estimated in 340–805 Gelatine Digestion Units. The resulting permeate successfully underwent fermentation for bioethanol production.

Platform design for extraction and isolation of Bromelain: Complex formation and precipitation with carrageenan

Abstract The main objective of this work was to investigate for the first time the molecular mechanism of complex formation between bromelain (a positively charged enzyme) and carrageenan (a natural strong polyelectrolyte, negatively charged) using spectroscopy techniques and thermodynamic approaches. The Bromelain-Carrageenan complex showed a maximal non-solubility at pH around 5.1. The solubility was dependent on pH and ionic strength of the medium. To re-dissolve the formed complex, the pH was changed and 500 mM of NaCl was added to the initial solution, proving the columbic mechanism for the formation of non-soluble complex. The formation of the carrageenan-bromelain complex increased in 8 °C the enzyme thermal stability, while its biological activity was not modified. The amount of total enzyme recovered in solution after precipitation with around 0.08% w/v of carrageenan was 85–90%.

The biochemical characterization, stabilization studies and the antiproliferative effect of bromelain against B16F10 murine melanoma cells

The current study aims to extract bromelain from different parts (stem, crown, peels, pulp and leaves) of Ananas comosus var. comosus AGB 772; to determine of optimum pH and temperature; to test bromelain stability in disodium EDTA and sodium benzoate, and to investigate its pharmacological activity on B16F10 murine melanoma cells in vitro. The highest enzymatic activity was found in bromelain extracted from the pulp and peel. The optimum bromelain pH among all studied pineapple parts was 6.0. The optimum temperature was above 50 °C in all bromelain extracts. The fluorescence analysis confirmed the stability of bromelain in the presence of EDTA and sodium benzoate. Bromelain was pharmacologically active against B16F10 melanoma cells and it was possible verifying approximately 100% inhibition of tumor cell proliferation in vitro. Since bromelain activity was found in different parts of pineapple plants, pineapple residues from the food industry may be used for bromelain extraction.

Recovery of bromelain from pineapple stem residues using aqueous micellar two-phase systems with ionic liquids as co-surfactants

Pineapple processing industries produce a large amount of residues; for example, the pineapple stem, which is rich in bromelain. Bromelain is a group of cysteine proteinases that have major importance in the pharmaceutical field due to, among others, their anti-inflammatory, antithrombotic, and fibrinolytic activities. Since Brazil is one of the largest producers of pineapples in the world, the development of an ecofriendly and cost-effective process to extract and purify bromelain from food/fruit residues would be significant. In this study, aqueous micellar two-phase systems composed of ionic liquids as co-surfactants were evaluated in the extraction of bromelain from the pineapple stem crude extract. The main results showed that bromelain partitions preferentially toward the micelle-poor phase, with enzyme recoveries above 90% for the studied systems. Moreover, a stabilizing effect of the ionic liquids toward the enzyme was observed.

AKTIVITAS ANTIBAKTERI DAN ANTIOKSIDAN HIDROLISAT HASIL HIDROLISIS PROTEIN SUSU KAMBING DENGAN EKSTRAK KASAR BROMELIN

Goat milk is highly nutritious foodstuffs that beneficial for improving health. The milk contains bioactive peptides which produced by hydrolysis process. The aim of this study was to evaluate antibacterial and antioxidant activities of hydrolisate produced from hydrolysis of goat milk protein by crude bromelain extract. Hydrolysis of goat milk protein was conducted using crude bromelain (0.1 U/mL) at pH 6, 50°C for 60 min. Hydrolysate was fractionated by using membrane molecular weight cut off 10 kDa. hydrolysate before and after fractionation were assayed for antibacterial and antioxidant activities. Toxicity of the Hydrolysate was determined by hemolysis assay. The result showed that the hydrolysate before and after fractionation inhibited growth of E. coli, S. Typhimurium and L. monocytogenes. Hydrolysate after fractionation has higher antibacterial activity indicated that fractionation was able to improve antibacterial activities of the hydrolysate fraction. The hydrolysate showed scavenging activity to ABTS and DPPH radicals. Fraction 10 kDa not only showed absence of hemolysis but also they were able to reduce autolysis of red blood cells. The result showed that hydrolysate from goat milk hydrolyzed by bromelain were able to be antibacterial and antioxidant.

Reverse micellar extraction of bromelain from pineapple peel – Effect of surfactant structure

Pineapple peel is generally disposed or used as compost. This study was focused on extracting bromelain from pineapple peel by using reverse micelles. It was found that gemini surfactant C12-8-C12·2Br (octamethylene-α,ω-bis(dimethyldodecylammonium bromide)) showed distinctive advantage over its monomeric counterpart DTAB (dodecyl trimethyl ammonium bromide); under optimized condition, the bromelain extracted with C12-8-C12·2Br reverse micelle had an activity recovery of 163% and a purification fold of 3.3, while when using DTAB reverse micelle, the activity recovery was 95% and the purification fold was 1.7. Therefore, the spacer of gemini surfactant should play a positive role in bromelain extraction and may suggest the potential of gemini surfactant in protein separation since it has been so far rarely used in relative experiments or technologies.

The effects of bromelain on angiogenesis, nitric oxide, and matrix metalloproteinase-3 and -9 in rats exposed to electrical burn injury

Abstract This study was aimed to investigate the effects of bromelain on angiogenesis, nitric oxide, and matrix metalloproteinase-3 and -9 in rats exposed to 1200 mV electrical burn injury. Thirty-five, male Wistar albino rats was divided into five groups (n = 7 each), including control (untreated) group; electrical burn injury (EI) group; electrical burn injury + 21.25 mg/kg BW bromelain group (EIB1); electrical burn injury + 42.50 mg/kg BW bromelain group (EIB2); and electrical burn injury + 85.00 mg/kg BW bromelain group (EIB3). Rat models of electrical burns done by providing electricity in rats that had been anesthetized, a voltage of 1200 mV and strong currents 15 mA for 10 s. The VEFG, NO, and MMP-9 levels were significantly greater in the EI group compared to the untreated group. Out of the 21.25 mg/kg BW, 42.50 mg/kg BW, and 85 mg/kg BW doses of Bromelain extract, only the lowest doses prevented EI-induced increase in NO and MMP-9 level (P

Application of an aqueous two‐phase micellar system to extract bromelain from pineapple (Ananas comosus) peel waste and analysis of bromelain stability in cosmetic formulations

Bromelain is a set of proteolytic enzymes found in pineapple (Ananas comosus) tissues such as stem, fruit and leaves. Because of its proteolytic activity, bromelain has potential applications in the cosmetic, pharmaceutical, and food industries. The present study focused on the recovery of bromelain from pineapple peel by liquid-liquid extraction in aqueous two-phase micellar systems (ATPMS), using Triton X-114 (TX-114) and McIlvaine buffer, in the absence and presence of electrolytes CaCl2 and KI; the cloud points of the generated extraction systems were studied by plotting binodal curves. Based on the cloud points, three temperatures were selected for extraction: 30, 33, and 36°C for systems in the absence of salts; 40, 43, and 46°C in the presence of KI; 24, 27, and 30°C in the presence of CaCl2 . Total protein and enzymatic activities were analyzed to monitor bromelain. Employing the ATPMS chosen for extraction (0.5 M KI with 3% TX-114, at pH 6.0, at 40°C), the bromelain extract stability was assessed after incorporation into three cosmetic bases: an anhydrous gel, a cream, and a cream-gel formulation. The cream-gel formulation presented as the most appropriate base to convey bromelain, and its optimal storage conditions were found to be 4.0 ± 0.5°C. The selected ATPMS enabled the extraction of a biomolecule with high added value from waste lined-up in a cosmetic formulation, allowing for exploration of further cosmetic potential.

Process optimization for reverse micellar extraction of stem bromelain with a focus on back extraction

In the current study, reverse micellar extraction (RME) for the purification of stem bromelain was successfully achieved using the sodium bis(2-ethylhexyl) sulfosuccinate (AOT)/isooctane system. A maximum forward extraction efficiency of 58.0% was obtained at 100 mM AOT concentration, aqueous phase pH of 8.0 and 0.2 M NaCl. Back extraction studies on altering stripping phase pH and KCl concentration, addition of counter-ion and iso-propyl alcohol (IPA) and mechanical agitation with glass beads indicated that IPA addition and agitation with glass beads have significant effects on extraction efficiency. The protein extraction was higher (51.9%) in case of the IPA (10% v/v) added system during back extraction as compared to a cetyltrimethylammonium bromide (100 mM) added system (9.42%). The central composite design technique was used to optimize the back extraction conditions further. Concentration of IPA, amount of glass beads, mixing time, and agitation speed (in rpm) were the variables selected. IPA concentration of 8.5% (v/v), glass bead concentration of 0.6 (w/v), and mixing time of 45 min at 400 rpm resulted in higher back extraction efficiency of 45.6% and activity recovery of 88.8% with purification of 3.04-fold. The study indicated that mechanical agitation using glass beads could be used for destabilizing the reverse micelles and release of bromelain back into the fresh aqueous phase.

Polymer‐based alternative method to extract bromelain from pineapple peel waste

Bromelain is a mixture of proteolytic enzymes present in all tissues of the pineapple (Ananas comosus Merr.), and it is known for its clinical therapeutic applications, food processing, and as a dietary supplement. The use of pineapple waste for bromelain extraction is interesting from both an environmental and a commercial point of view, because the protease has relevant clinical potential. We aimed to study the optimization of bromelain extraction from pineapple waste, using the aqueous two-phase system formed by polyethylene glycol (PEG) and poly(acrylic acid). In this work, bromelain partitioned preferentially to the top/PEG-rich phase and, in the best condition, achieved a yield of 335.27% with a purification factor of 25.78. The statistical analysis showed that all variables analyzed were significant to the process.