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Characterization of cross-linked immobilized arylesterase from Gluconobacter oxydans 621H with activity toward cephalosporin C and 7-aminocephalosporanic acid

机译:从葡聚糖酸葡萄糖C和7-氨基孢菌酸的葡聚糖氧化物621H中交联固定芳基酯酶的表征

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Cross-linked enzyme aggregates (CLEAs) were prepared from several precipitant agents using glutaraldehyde as a cross-linking agent with and without BSA, finally choosing a 40% saturation of ammonium sulfate and 25mM of glutaraldehyde. The CLEAs obtained under optimum conditions were biochemically characterized. The immobilized enzyme showed higher thermal activity and a broader range of pH and organic solvent tolerance than the free enzyme. Arylesterase from Gluconobacter oxydans showed activity toward cephalosporin C and 7-aminocephalosporanic acid. The CLEAs had a Kcat/K-M of 0.9M(-1)/S-1 for 7-ACA (7-aminocephalosporanic acid) and 0.1M(-1)/S-1 for CPC (cephalosporin c), whereas free enzyme did not show a typical Michaelis-Menten kinetics. (c) 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:36-42, 2016
机译:使用戊二醛作为具有和不具有BSA的交联剂的戊二醛作为交联剂的几种沉淀剂制备交联酶聚集体(CLEA),最终选择40%的硫酸铵和25mm的戊二醛饱和。 在最佳条件下获得的CLEA生物化学表征。 固定化酶显示出比游离酶更高的热活性和更广泛的pH和有机溶剂耐受性。 来自葡糖的芳基酯酶onydans显示出对头孢菌素C和7-氨基孢菌酸的活性。 本叶片具有0.9m(-1)/ km的kcat / km,用于7-aca(7-氨基氨基磺酸)和0.1m(-1)/ s-1的CPC(头孢菌素C),而游离酶 不显示典型的michaelis-menten动力学。 (c)2015美国化学工程研究所生物科技。 Prog。,32:36-42,2016

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