• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biotechnology Progress >Conjugation of Penicillin Acylase with the Reactive Copolynmer of N-Isopropylacrylamide: A Step Toward a Thermosensitive Industrial Biocatalyst
【24h】

Conjugation of Penicillin Acylase with the Reactive Copolynmer of N-Isopropylacrylamide: A Step Toward a Thermosensitive Industrial Biocatalyst

机译：青霉素酰化酶与N-异丙基丙烯酰胺反应性共聚体的缀合：迈向热敏工业生物催化剂的一步

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Conjugation of penicillin acylase (PA) to poly-N-isopropylacrylamide (polyNIPAM) was studied as a way to prepare a thermosensitive biocatayst for industrial applications to antibiotic synthesis. Condenation of PA with the copolymer of NIPAM containing active ester groups resulted in higher coupling yields of the enzyme (37%) compared toits chemical modification and copolymerization with the monomer (9% coupling yield) at the same NIPAM: enzyme weight ratio of ca. 35. A 10-fold increase of the enzyme loading on the copolymer resulted in 24% coupling yield and increased by 4-fold the specific PA activity of the conjugate. Two molecular forms of the conjugate were found by gel filtratio non Sepharose CL 4B: the lower molecular weight fraction of ca. 10~6 and, presumably, cross-linked protein-polymer aggregates of MW > 10~7. Michaelis constant for 5-nitro-3-phenylacetamidobenzoic acid hydrolysis by the PA conjugate (20 muM) was found to be slightly higher than that of the free enzyme (12 muM), and evaluated of V_max testifies to the high catalytic efficiency of the conjugated enzyme. PolyNIPAM-cross-linked PA retained its capacity to synthesize cephalexin from D-phenylglycin amide and 7-aminodeacetoxycephalosporanic acid. The synthesis-hydrolysis ratios of free and polyNIPAM-cross-linked enzyme in cephalexin synthesis were 7.46 and 7.49, respectively. Thus, diffusional limitation, which is a problem in the industrial production of beta-lactam antibiotics, can be successfully eliminated by cross-linking penicillin acylae to a smart polymer (i.e., polyNIPAM).
机译：研究了将青霉素酰化酶（PA）与聚N-异丙基丙烯酰胺（polyNIPAM）结合的方法，以制备用于工业化合成抗生素的热敏生物催化剂。在相同的NIPAM：酶的重量比约为10的情况下，PA与含有活性酯基团的NIPAM的共聚物缩合可导致酶的偶联率更高（37％），与化学改性和与单体的共聚反应（9％偶联率）相比。 35.共聚物上酶负载的10倍增加导致偶联收率提高24％，共轭物的比PA活性提高4倍。通过凝胶过滤非琼脂糖凝胶CL 4B发现了两种分子形式的结合物：分子量较低的ca。 10〜6，可能是MW> 10〜7的交联蛋白-聚合物聚集体。发现PA共轭物（20μM）水解5-硝基-3-苯基乙酰氨基苯甲酸的米氏常数比游离酶（12μM）略高，并且对V_max的评估证明了共轭物的高催化效率酶。 PolyNIPAM交联的PA保留了从D-苯基甘氨酸酰胺和7-氨基脱乙酰氧基头孢菌素酸合成头孢氨苄的能力。头孢氨苄合成中游离和聚NIPAM交联酶的合成水解率分别为7.46和7.49。因此，通过将青霉素酰基转移酶交联到智能聚合物（即polyNIPAM）上，可以成功消除在β-内酰胺抗生素的工业生产中存在的问题的扩散限制。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号