Jack Bean Meal Research Articles

Heteroatom self-doped hierarchical porous carbon from nitrogen-rich jack bean meal for high-performance supercapacitor and efficient oxygen reduction

Heteroatom self-doped hierarchical porous carbon from nitrogen-rich jack bean meal for high-performance supercapacitor and efficient oxygen reduction

The impact of enzyme addition on the metabolizable energy and protein digestibility of peeled Jack bean meal (Canavalia ensiformis L.)

The impact of enzyme addition on the metabolizable energy and protein digestibility of peeled Jack bean meal (<i>Canavalia ensiformis</i> L.)

Investigating purification and activity analysis of urease enzyme extracted from jack bean source: A green chemistry approach

Urease is an enzyme of historical importance in the field of biochemistry, generally microbial and plant urease is the primary sources of urease. The significant applications of urease enzyme are found to be foremost in food industry, medical equipment's and biosensors. In this work, urease has been extracted from Jack bean meal using ammonium sulphate and acetone precipitation. A significant amount of urease was precipitated and concentrated at 60% saturated solution of ammonium sulphate. The obtained precipitates were dissolved in 50 mM phosphate buffer (pH 8) after centrifugation, and subjected to sodium dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) to determine the molecular weight of urease. Results obtained from the SDS-PAGE were validated using Zymography. Anion exchange chromatography was used to separate the desired protein at different pH (7.0, 7.5 and 8.0). The eluted fractions were assessed for urease activity using phenol-nitroprusside dependent ammonia release assay. Under these assay conditions, one unit of urease activity was calibrated as the amount of enzyme liberating 1 μM of NH3 from urea per unit time. The eluted fraction and Zymography analysis show the purified urease observed at 90 kDa and activity of purified urease, respectively. The obtained results for specific activity (173.67Units mg) and % purification (99.71%) for urease has been compared with the available literature, which are found to be in close relation with existing results. The proposed method is a novel approach which has recorded highest % purification and specific activity. Furthermore, it can be suitable for extracting urease from jack bean source for various industrial applications.

Determination of the optimum dietary levels of cracked and cooked jackbean meal for finisher broilers

A 35-day feeding trial was conducted to determine the optimal dietary level(s) of cracked and cooked (CAC) jackbean meal for finisher broilers. Five diets were formulated such that they contained cracked and cooked jackbean meal ai 0%, 15%, 20%, 25% and 30% levels respectively. Each diet was fed to a group of 45 finisher broiler birds beginning at 5 weeks of age. Average daily feed intake of the groups were 99.00g, 95.00g, 89.00g, 89.00g and 90.00g for the control, 15%, 20%, 25% and 30% respectively. Body weight gains were 34.00g, 33.00g, 30.00g, 27.00g and 23.00g respectively and feed conversion ratio for the groups were 2.95, 2.88, 2.91, 3.25 and 3.83 respectively. There were no significant (P &gt;0.05) differences among the control (0%), 15% and 20% groups in feed intake, body weight gain and feed conversion ratio. The 25% und 30% groups however, showed significantly (P &lt; 0.05) depressed body weight gain and poor feed conversion ratio. Organ weights of the birds relative to body weight did not indicate any significant differences among the five experimental groups. The results of this trial suggest that the optimal dietary inclusion of cracked and cooked jackbean meal for finisher broiler is 20%

Temperature‐dependent inactivation and catalysis rates of plant‐based ureases for engineered biomineralization

AbstractEngineered (bio)mineralization uses the enzyme urease to catalyze the hydrolysis of urea to promote carbonate mineral precipitation. The current study investigates the influence of temperature on ureolysis rate and degree of inactivation of plant‐sourced ureases over a range of environmentally relevant temperatures. Batch experiments at 30°C demonstrated that jack bean meal (JBM) has a 1.7 to 56 times higher activity (844 μmol urea hydrolyzed g−1 JBM min−1) than the other tested plant‐sourced ureases (soybean, pigeon pea and cottonseed). Hence, ureolysis and enzyme inactivation rates were evaluated for JBM at temperatures between 20°C and 80°C. A combined first‐order urea hydrolysis and first‐order enzyme inactivation model described the inactivation of urease over the investigated range of temperatures. The temperature‐dependent rate coefficients (kurea) increased with temperature and ranged from 0.0018 at 20°C to 0.0249 L g−1 JBM min−1 at 80°C; JBM urease became ≥50% inactivated in as little as 5.2 minutes at 80°C and in as long as 2238 minutes at 50°C. The combined urea hydrolysis kinetics and enzyme inactivation model provides a mathematical relationship useful for the design of biomineralization technologies and can be incorporated into reactive transport models.

Crude Urease Extract for Biocementation

A crude extract from jack beans (Canavalia gladiata) is demonstrated to be an effective source of urease enzyme for biocementation via enzyme-induced carbonate precipitation (EICP). Test tube tests of crude and purified extracts from jack beans, jack bean meal, soybeans, and watermelon seeds show that the crude jack bean extract results in the highest unit yield, defined as urease content per initial mass of source material, among these four plant sources. The efficacy of EICP using crude jack bean extract for biocementation was compared with the efficacy of three commercially available enzymes by biocementation of a granular soil. Unconfined compression tests on the granular soil specimens subject to biocementation via EICP demonstrated that the crude extract and the less purified commercially available enzyme were actually more effective than commercially available highly purified urease enzymes at enhancing soil strength, an effect attributed to the presence of complementary proteins in the less purified enzyme sources. The simple technique used to produce the crude extract from jack beans significantly lowers the cost of EICP, eliminating a major barrier to practical applications, including infrastructure construction and environmental protection.

Guidance for Investigating Calcite Precipitation by Urea Hydrolysis for Geomaterials

Abstract Microbially induced calcite precipitation (MICP) is a sustainable method of stabilizing (i.e., cementing) loose sandy deposits or creating an impervious barrier within the soil mass or both. MICP can occur through various biochemical pathways, and ‘Urea Hydrolysis’ (UH) is considered to be the most efficient of the MICP methods for biochemically inducing calcite precipitation. To date, the geotechnical engineering community that has investigated MICP has tended to focus on the hydromechanical behavior of the end product, i.e., MICP cemented sands; however, many biochemical factors that affect reaction-rate kinetics and MICP outcomes have been understudied or neglected. This study investigates the kinetics of UH and compares different sources of the urease enzyme—those microbially cultivated in the laboratory (i.e., Sporosarcina pasteurii) and those extracted from plants (i.e., Jack bean meal)—to investigate the influence of urea concentration, buffer capacity, and the cell harvesting method on UH. Through this study, an attempt has been made to arrive at an optimal concentration of urea, under the influence of the previously mentioned parameters and the buffering action of the soil, on urea hydrolysis. These results have implications for optimizing MICP and, in particular, for upscaling these methods to in situ applications.

Utilization of Jack Bean (Canavalia ensiformis) Meal as a Substitute for Soybean Meal in Diet for Broiler Reared for 35 Days

This study was aimed to evaluate the utilization of jack bean meal as a substitution of soybean meal in the diets and the effect on broiler performance. A total number of two hundred Lohmann MB 202 Platinum broiler chickens were kept in five weeks rearing period, consisted of three weeks of starter phase (0-3 weeks) and two weeks of finisher phase (3-5 weeks). This study used a completely randomized design with four treatments and five replicates. The dietary treatments were: JB0-0 (control ration: basal diet without jack bean meal substitution in starter and in finisher phase), JB50-0 (basal diet with jack bean meal substitution in starter phase only), JB50-50 (basal diet with jack bean meal substitution in starter and in finisher phase), JB0-50 (basal diet with jack bean meal substitution in the finisher phase only). The data obtained were statistically analyzed using analysis of variance and followed with Duncan's multiple-range test for the data with significant difference. The variables measured were: growth performance, carcase yield, visceral organ weight and immune organ. Results showed that the dietary treatment had no effect on performance, carcass yield and commercial cut, visceral organ or immune organs of 35 days old broiler chickens, except for the thymus (P&lt;0.05). It can be concluded that as a source of protein, soybean meal can be replaced by jack bean meal up to 50%. Jack bean meal can be given either in starter phase only, in finisher phase only, or in both starter and in finisher phase.

Nutritional value of raw Canavalia ensiformis and its utilization as partial replacement for soybean meal in the diet of Clarias gariepinus (Burchell, 1822) fingerlings.

The nutritional value of raw Jack bean meal (Canavalia ensiformis) as a partial substitute for soybeans meal was investigated in this study. Preliminary investigation on nutrient composition revealed that lysine, histidine, and phenylalanine were significantly higher in C. ensiformis seed meal compared to soybean meal. However, crude protein and other essential amino acids were significantly lower. Feeding trial was then conducted to investigate the effect of replacing about 40% soybeans meal (at 58.8% inclusion) with C. ensiformis in the diet of Clarias gariepinus. The result obtained after 56 days revealed that fingerlings could tolerate up to 20% replacement without significant effect on growth and nutrient utilization. Beyond this, growth was significantly reduced. Survivals of the fish also follow a similar trend as stated above. It was concluded that dietary inclusion of raw C. ensiform meal should not be beyond 11% (or 20% replacement for soybeans meal included at 58.8%) in the diet of C. gariepinus.

Evaluation of the Determination of Free Urea in Water-Soluble Liquid Fertilizers Containing Urea and Ureaforms by Urease and HPLC Methods.

Currently there are three AOAC Official Methods for the determination of urea in fertilizers. AOAC Official Method 959.03, Urea in Fertilizers, Urease Method, First Action 1959, Final Action 1960, is based on the use of fresh commercial 1% urease solution, or preparation of such solution from urease powder in water, or from jack bean meal in water. AOAC Official Method 983.01, Urea and Methyleneureas (Water-Soluble) in Fertilizers, First Action 1983, Final Action 1984, is based on LC with a refractive index detector using water as the mobile phase and a C18 column. AOAC Official Method 2003.14, Determination of Urea in Water- Soluble Urea-Formaldehyde Fertilizer Products and in Aqueous Urea Solutions, First Action 2003, Final Action 2008, is based on LC with a UV detector using acetonitrile-water (85+15, v/v) mobile phase and a propylamine column. The urea method, AOAC Official Method 959.03, is very much dependent on the nature of the urease enzyme. The method was developed in 1960 and used for simple urea fertilizer solutions. With the advent of complex fertilizer compositions, especially with the class of liquid triazone fertilizers and water-soluble urea forms, the analyses of free urea in these fertilizers by the urease method is often inaccurate and inconsistent. AOAC Official Method 983.01 is not always reliable due to the interference of some of the components of these fertilizers, and due to the fact that the use of water as the mobile phase does not always separate the free urea from other components. AOAC Official Method 2003.14 was subjected to ring test studies that showed it could be used for the determination of "free urea" in these classes of fertilizers with good accuracy and precision.

Effect of additives on the purification of urease

The effect of additives on the purification of proteins was investigated. The target protein studied here is the enzyme urease. Studies on the purification of urease from jack bean meal were carried out. 32% (v/v) acetone was utilized to extract urease from the jack bean meal. Further purification by crystallization with the addition of 2-mercaptoethanol and EDTA disodium salt dehydrate was carried out. It was found out that the presence of additives can affect the selectivity of the crystallization. Increases in both purity and yield of the urease after crystallization were observed in the presence of additives, which were proven using both SDS-PAGE and activity. Urease crystals with a yield of 69.9% and a purity of 85.1% were obtained in one crystallization step in the presence of additives. Furthermore, the effect of additives on the thermodynamics and kinetics of urease crystallization was studied.

Solvent-freeze-out (SFO) technology: A controlled crystallization process—Case study of jack bean urease

The solvent freeze out (SFO) technique is here the first time introduced and used to purify a realistic and complex protein mixture. Urease crystals were obtained from jack bean meal in the process and the purity of which were evaluated via both specific activity and SDS-PAGE. Compared to a conventional salting out crystallization, the use of SFO technology can provide a better control of the crystallization process. A detailed investigation into the crystallization process is presented by carrying out experiments with different parameters. Results showed that the quality of the ice layer is the key factor for the control of the crystallization process. The SFO technology can be considered a promising alternative in protein crystallization, because the process itself is efficient and reproducible. Even though not fully optimized, a satisfactory product purity and a decent yield of urease crystals were obtained using the SFO technology. The enzymatic activity was also preserved after the SFO process in the here carried out case study.

Production and characterization of a camelid single domain antibody-urease enzyme conjugate for the treatment of cancer.

A novel immunoconjugate (L-DOS47) was developed and characterized as a therapeutic agent for tumors expressing CEACAM6. The single domain antibody AFAIKL2, which targets CEACAM6, was expressed in the Escherichia coli BL21 (DE3) pT7-7 system. High purity urease (HPU) was extracted and purified from Jack bean meal. AFAIKL2 was activated using N-succinimidyl [4-iodoacetyl] aminobenzoate (SIAB) as the cross-linker and then conjugated to urease. The activation and conjugation reactions were controlled by altering pH. Under these conditions, the material ratio achieved conjugation ratios of 8-11 antibodies per urease molecule, the residual free urease content was practically negligible (<2%), and high purity (>95%) L-DOS47 conjugate was produced using only ultradiafiltration to remove unreacted antibody and hydrolyzed cross-linker. L-DOS47 was characterized by a panel of analytical techniques including SEC, IEC, Western blot, ELISA, and LC-MS(E) peptide mapping. As the antibody-urease conjugate ratio increased, a higher binding signal was observed. The specificity and cytotoxicity of L-DOS47 was confirmed by screening in four cell lines (BxPC-3, A549, MCF7, and CEACAM6-transfected H23). BxPC-3, a CEACAM6-expressing cell line was found to be most susceptible to L-DOS47. L-DOS47 is being investigated as a potential therapeutic agent in human phase I clinical studies for nonsmall cell lung cancer.

Development Of Nanobiosensor For Urea By Immobilizing Jack Bean Meal Urease On CdS QDs

Semiconductor nanoparticles (NPs) have attracted much attention as a new class of fluorescent probe for many biological applications including biosensors for glucose, cholesterol, cysteine etc., which were based on their size dependent unique electrical and photophysical properties. One of the major challenges in fabricating nanomaterial based biosensor is conjugation of a suitable compound to quantum dots which is preferably selective to the analyte of interest. Here we report our studies on synthesis, characterization and application of jack bean meal urease immobilized on CdS quantum dots (QDs) for sensing of urea. The CdS QDs were synthesized by chemical precipitation method using Mercaptoacetic acid (MAA) for controlling size as well as for imparting functional group for conjugating urease. The urease immobilize on MAA capped CdS nanoparticle was characterized by an array of techniques, like, UV- visible, Fluorescence, XRD, FT-IR and SEM EDAX. The detection capability of urea was studied by fluorescence spectroscopy at an excitation wavelength of λex = 430 nm and emission wavelength of λem= 546 nm. This method was capable to detect urea in the concentration range of 0.1 µM to 1 mM. Copyright © 2013 VBRI press.

Kinetics of urease mediated calcite precipitation and permeability reduction of porous media evidenced by magnetic resonance imaging

The enzyme urease drives the hydrolysis of urea leading to the release of ammonium ions and bicar- bonate; in the presence of calcium, the rise in pH leads to increased calcium carbonate saturation and the subsequent precipitation of calcite. Although such alkalinizing ureol- ysis is widespread in nature, most studies have focussed on bacteria (i.e. indigenous communities or urease-active Sporosarcina pasteurii) for calcite precipitation technolo- gies. In this study, urease-active jack bean meal (from the legume Canavalia ensiformis) was used to drive calcite precipitation. The rates of ureolysis (kurea), determined from measured NH4 ? , enabled a direct comparison to microbial ureolysis rates reported in literature. It is also demonstrated that a simple single reaction model approach can simulate calcite precipitation very effectively (3-6 % normalised root-mean-square deviation). To investigate the reduction of permeability in porous media, jack bean meal (0.5 g L -1 ) and solutions (400 mM urea and CaCl2) were simultaneously pumped into a borosilicate bead column. One-dimensional magnetic resonance profiling techniques were used, non-invasively, for the first time to quantify the porosity changes following calcite precipitation. In addi- tion, two-dimensional slice selective magnetic resonance images (resolution of *0.5 9 1.0 mm) revealed that the exact location of calcite deposition was within the first 10 mm of the column. Column sacrifice and acid digestion also confirmed that 91.5 % of calcite was located within the first 14 mm of the column. These results have impor- tant implications for the design of future calcite precipi- tation technologies and present a possible alternative to the well known bacterial approaches.

Purification of urease from jack bean (Canavalia ensiformis) with copper (II) chelated poly(hydroxyethyl methacrylate‐N‐methacryloyl‐(l)‐histidine methyl ester) cryogels

Jack bean (Canavalia ensiformis) is the source of interesting proteins that contribute to modern biochemistry, and urease is the primary of these proteins. Owing to its role and occurrence in nature, urease has become a part of extensive studies. In this study, jack bean urease (JBU) was purified by immobilized metal affinity chromatography using Cu(2+) chelated poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester) [PHEMAH-Cu(2+)]-based cryogels. PHEMAH-Cu(2+) cryogel was synthesized and characterized for swelling degree, morphology (by SEM), N-methacryloyl-(L)-histidine methyl ester and Cu(2+) incorporation (by elemental analysis and atomic absorption spectrophotometry). The binding of JBU to PHEMAH-Cu(2+) cryogel was optimized by examining the effect of pH, flow rate and JBU concentration on binding. The maximal binding of JBU was 23.2 mg/dry gram of adsorbent. The maximal binding of JBU extracted from jack bean meal was 67.8 mg/dry gram of adsorbent. The elution of JBU from cryogel column was accomplished by 1.0 M NaCl in 20 mM phosphate buffer (pH 8.0). Molecular weight and purity of JBU from jack bean meal was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was observed that JBU could be repeatedly bound and eluted from (PHEMAH)-Cu(2+) cryogel with less than 10% loss in column capacity.

Growth and Nutrient Utilization of Clarias gariepinus Fed Dietary Levels of Jackbean (Canavalia ensiformis) Meal

Clarias gariepinus fingerlings of average weight 0.71 g were sourced from ARAC, Port Harcourt and used to evaluate the effect of feeding dietary levels of Jackbean meal on growth and nutrient utilization of the fish. 35% isonitrogenous diets of 0%, 10%, 20% and 30% dietary levels of JBM were formulated. These were fed to the fingerlings randomly assigned to 4 treatments - TCN (Control), TJ1 (10% JBM), TJ2 (20% JBM) and TJ3 (30% JBM) respectively in 3 replicates of 13 fingerlings each, using 12 plastic aquaria of 250 x 150 cm dimension. The fish were fed at 5% body weight twice daily within the experimental period of 8 weeks. Mortality was highest in TJ3 and least in the control (TCN). Daily feed intake and protein intake were not significantly (p>0.05) different. The control (TCN) had the highest body weight gain, followed by TJ3, TJ1 and TJ2. Specific growth rate and feed conversion ratio of the treatments were not significantly (p>0.05) different. The protein efficiency ratio of treatment TJ3 was significantly (p<0.05) higher than the rest of the treatments. Dietary levels of JBM up to 30% inclusion level therefore had nutritional attributes as feedstuff in the diet of Clarias gariepinus fingerlings.

Digestibility coefficients of processed jackbean meal Cannavalia ensiformis (L.) DC for Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) diets

A study was conducted to determine in-vivo apparent digestibility coefficient of nutrients in processed jackbean meal. Acid Insoluble Ash (AIA) was used as an indicator. The test diets consist of the seed meal replacing soybean meal at 10, 20, 30 and 40% to serve as test diets D10, D20, D30, and D40 respectively. A diet called as diet CTR, without jackbean meal served as control. These diets were fed to Tilapia Oreochromis niloticus fingerlings (6.37 ± 0.07 g) in a glass tanks for 5 weeks. Faecal collection was made by siphoning after four hours of each feeding. Proximate analysis of both diets and faecal samples were carried out for crude protein, lipid, crude fiber, energy, and AIA. These were used to calculate the digestibility coefficient of nutrients. There was no significant difference (p > 0.05) in lipid and organic matter digestibility of the fish fed control diet and test diets. Significant (p < 0.05) variations occurred in nitrogen-free extracts, protein and crude fiber digestibility of fish fed control diet and the test diets. Key words: Digestibility, in vivo, jackbean, Oreochromis niloticus.

Immobilization of urease in alginate, paraffin and lac

The enzyme urease (EC.3.5.1.5) from jack bean meal was immobilized by various techniques, such as entrapment in calcium alginate gel spheres in aqueous suspension, lac impregnated muslin cloth as dry films and by embedding in paraffin wax impregnated muslin cloth. The activity of the free and immobilized enzymes as a function of pH, temperature, storage stability, kinetic parameters and periodic use were compared. The immobilized enzyme showed good storage stability. After repeated use, the alginate beads turned brown and deteriorated, hence the storage stability was not good. The paraffin films were preserved dry because during wet preservation, the film slightly softened and the protein leached out slightly. The alginate beads had moderate mechanical stability. The lac films were tougher than the paraffin wax films in terms of mechanical stability. The Km and Vmax values were altered after immobilization. The Km values for calcium alginate and lac were low, while it was larger in paraffin film as compared to the free enzyme. This may be due to the fact that immobilization on calcium alginate and lac in presence of CaCl2 and methanol exposed certain active sites of the urease. While immobilization on paraffin masks the active sites and may lead to reduced binding of the substrate.

Purification, crystallization and preliminary X-ray analysis of urease from jack bean (Canavalia ensiformis)

Plant urease is a seed protein that is common in most legumes. It is also common in many bacteria and fungi and several species of yeast. Urease allows organisms to use exogenous and internally generated urea as a nitrogen source by catalyzing the hydrolysis of urea to ammonia and carbon dioxide. Urease from jack bean meal was purified to electrophoretic homogeneity using a series of steps involving acetone precipitation and size-exclusion and ion-exchange chromatography. The jack bean urease was crystallized and the resulting crystals diffracted to 2.05 A resolution using synchrotron radiation. The crystals belonged to the hexagonal space group P6(3)22, with unit-cell parameters a = b = 138.57, c = 198.36 A.