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

Abstract

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.

Synthesis of molecularly imprinted magnetic nanoparticles for selective cytidine adsorption

“…The adsorption amount of the template molecule significantly decreased at lower and higher pH values, probably due to charge repulsion. Here, the electrostatic interaction at pH 7.4 corresponds to the interaction between negatively charged cytidine and positively charged Cu(II) ions [26,29].…”

Molecularly imprinted cryogel columns for Concanavalin A purification from jack bean extract

“…Jack bean (Canavalia ensiformis) seeds were ground into flour [11]. 25 g jack bean flour has been left with a magnetic mixer in a pH 8.0 Tris HCI buffer during the night.…”

“…ln e = ln f +1∕nln e (11) where b is the Langmuir isotherm constant, K f is the Freundlich constant and n is the Freundlich exponent. The value of 1/n ranges between 0 and 1 and it is a measure of surface heterogeneity, becoming more heterogeneous as its value gets closer to zero [34].…”

“…Con A, isolated from the jack bean, has neutral pI, with a molecular mass of 104 kDa. Con A from Canavalia ensiformis (C. ensiformis) is a tetramer with four carbohydrate binding sites which usually binds to specific saccharides such as mannose and glucose [11][12][13][14]. For this reason, it requires divalent cations (i.e.…”

Multiphysics model development to characterize fundamental mechanism of bio-responsive hydrogels

“…Thereby, the plant-based ureases were usually obtained from jack bean [55], soybean [56] and pigeon pea [57], while the bacterial-origin ureases were typically extracted from Bacillus pasteurii [58,59], Proteus mirabilis [60] and Providencia rettgeri [56]. To improve the yield of the enzyme, urease was typically purified, increasing the urease catalytic activity by either: (i) crystallization [61] or (ii) chromatographic [62]. In order to synthesize the urea-sensitive urease-loaded hydrogel, researchers employed several techniques for the immobilization of urease in the hydrogel, such as: (i) covalent binding [63], (ii) adsorption [64], (iii) encapsulation [65], and (iv) entrapment [66].…”