02723nas a2200325   4500008004100000022001400041245019400055210006900249260007300318300001200391490000700403520146800410653003301878653001501911653002601926653002501952653002501977653001702002653003502019653002602054653001202080100001802092700003102110700002002141700002402161700002902185700003002214700002502244856012802269       2012                        eng d  a0927-776500aAdsorption induced enzyme denaturation: the role of protein surface in adsorption induced protein denaturation on allyl glycidyl ether (AGE)-ethylene glycol dimethacrylate (EGDM) copolymers0 aAdsorption induced enzyme denaturation the role of protein surfa  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcFEB  a184-1900 v903 a<p>The effects of protein size on adsorption and adsorption-induced denaturation of proteins on copolymers of allyl glycidyl ether (AGE)-ethylene glycol dimethacrylate (EGDM) have been studied. Different responses were observed for the amount of protein adsorbed and denatured on the polymer surface for different proteins (trypsin, alchol dehydrogenase from baker's yeast (YADH), glucose dehydrogenase (GDH) from Gluconobacter cerinus, and alkaline phosphates from calf intestinal mucosa (CIAP). Protein adsorption on the copolymer with 25% crosslink density (AGE-25) was dependent not only on the size of the protein but also on the presence of glycoside residues on the protein surface. Adsorption and denaturation of proteins follows the order YADH &gt; trypsin &gt; GDH &gt;&gt; CIAP although the molecular weights of the proteins follow the order YADH &gt; CIAP &gt; GDH &gt; trypsin. The lack of correlation between amount of adsorbed protein and its molecular weight was due to the presence of glycoside residues on CIAP and GDH which protect the enzyme surface from denaturation. Enzyme stabilities in aqueous solutions of 1-cyclohexyl-2-pyrrolidinone (CHP) correlate well with the trend in denaturation by the copolymer, strongly suggesting that hydrophobic interactions play a major role in protein binding and the mechanism of protein denaturation is similar to that for water-miscible organic solvents. (C) 2011 Elsevier B.V. All rights reserved.</p>10a1-Cyclohexyl-2-pyrrolidinone10aAdsorption10aAlcohol dehydrogenase10aAlkaline phosphatase10aallyl glycidyl ether10aDenaturation10aethylene glycol dimethacrylate10aGlucose dehydrogenase10aTrypsin1 aThudi, Lahari1 aJasti, Lakshmi, Swarnalath1 aSwarnalatha, Y.1 aFadnavis, Nitin, W.1 aMulani, Khudbudin, Baban1 aDeokar, Sarika, Babasaheb1 aPonrathnam, Surendra  uhttps://library.ncl.res.in/content/adsorption-induced-enzyme-denaturation-role-protein-surface-adsorption-induced-protein-003073nas a2200289   4500008004100000022001400041245010500055210006900160260007300229300001000302490000700312520198800319653002502307653002302332653003502355653002602390653002102416653003202437100001802469700003102487700002502518700002402543700002902567700003002596700002502626856013202651       2012                        eng d  a1381-117700aEnzyme immobilization on epoxy supports in reverse micellar media: prevention of enzyme denaturation0 aEnzyme immobilization on epoxy supports in reverse micellar medi  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcJAN  a54-620 v743 a<p>Immobilization of enzymes such as alpha-chymotrypsin (EC 3.4.21.1), yeast alcohol dehydrogenase (YADH) from Saccharomyces cerevisiae (EC 1.1.1.1) and glucose dehydrogenase (GDH) from Gluconobacter cerinus (EC 1.1.1.119) has been carried out. Copolymers of allyl glycidyl ether (AGE) crosslinked with 25% ethylene glycol dimethacrylate (EGDM) (25 mg, dry wt) were contacted with the enzymes solubilized in reverse micellar media (0.5-5 mg/mL)(overall) of sodium bis(2-ethylhexyl) sulfosuccinate (AOT) salt in isooctane, and cetyl trimethylammonium bromide (CTAB) in chloroform-isooctane (50:50, v/v). Although the enzymes are readily denatured (&gt;90%) after adsorption on the copolymer in aqueous buffers, no such adsorption-induced denaturation takes place in reverse micelles. alpha-Chymotrypsin is remarkably stable in AOT reverse micelles when 0.025 M citrate buffer of pH 9.0 containing 2 mM CaCl2 is used in the water pools instead of Tris-HCl buffer of pH 8.5. It was possible to achieve enzyme concentration of 5 mg/mL in 0.3 M AOT at molar ratio of water to surfactant, (W-0), 30 and to obtain alpha-chymotrypsin loading of 20 mg/g of copolymer. The recovered enzyme solution can be reused with a fresh batch of polymer after supplementing the depleted solution. The immobilized enzyme exhibits excellent stability in aqueous buffers at room temperature and can be recycled several times. YADH is stable in both AOT and CTAB reverse micelles while GDH is stable only in CTAB reverse micelles containing 0.05 M Tris-HCI buffer of pH 8.5. Interestingly, the combination of YADH (2.5 mg/g) and GDH (0.5 mg/g) co-immobilized on the copolymer using CTAB-chloroform-isooctane system can be used for regeneration and recycle of NADPH at least 50 times as exemplified by complete reduction of a prochiral ketoester, ethyl 4-phenyl-2,4-dioxobutyrate (10 mM) to ethyl (R)-2-hydroxy-4-phenylbutyrate (HPB ester) using NADPH (0.2 mM). (C) 2011 Elsevier B.V. All rights reserved.</p>10aallyl glycidyl ether10aalpha-Chymotrypsin10aethylene glycol dimethacrylate10aGlucose dehydrogenase10aReverse micelles10aYeast alcohol dehydrogenase1 aThudi, Lahari1 aJasti, Lakshmi, Swarnalath1 aSwarnalatha, Yalangi1 aFadnavis, Nitin, W.1 aMulani, Khudbudin, Baban1 aDeokar, Sarika, Babasaheb1 aPonrathnam, Surendra  uhttps://library.ncl.res.in/content/enzyme-immobilization-epoxy-supports-reverse-micellar-media-prevention-enzyme-denaturation-0