固定化双酶耦联体系制备手性胺
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摘要
胺脱氢酶(amine dehydrogenase,Am DH)能够在辅酶的作用下不对称还原前手性酮制备手性胺,并与甲酸脱氢酶(formate dehydrogenase,FDH)构建耦联双酶反应体系,通过酶活检测与高效液相色谱法得出,Am DH和FDH浓度比为4∶1、底物浓度为10 mmol/L、辅酶浓度为0. 025 mmol/L时,是游离双酶耦联体系最优反应条件.为提高双酶耦联体系的稳定性,将聚乙烯亚胺-双酶复合物作为模板和催化剂,诱导钛前驱体Ti-BALDH(titanium(IV) bis-(ammoniumlactato)-dihydroxide)缩聚,形成固定化耦联体系.与游离酶体系相比,辅酶可在固定化酶体系的微环境中高效循环再生,大大提高了多酶耦联体系的催化效率.
Amine dehydrogenase( Am DH) can synthesize chiral amines by asymmetric reduction of prochiral ketones with coenzyme and establish a coupled double enzyme reaction system with formate dehydrogenase( FDH).The results of enzyme activity detection and high performance liquid chromatography show that the optimal reaction condition is that the concentration ratio of Am DH to FDH is 4 ∶ 1 and the concentration of substrate and the concentration of coenzyme are 10 mmol/L and 0. 025 mmol/L,respectively. In order to improve the stability of the system,the polyethylenimine( PEI) coating on the enzyme surface to form polyethyleneimine-diphenylase complex as the template and catalyst is employed for the condensation of Ti-BALDH to form immobilized double enzyme coupling system. Compared with the free double enzyme system,the coenzyme can be recycled and regenerated efficiently in the microenvironment of the immobilized enzyme system. It greatly improves the catalytic efficiency of the multi-enzyme coupling system.
Amine dehydrogenase( Am DH) can synthesize chiral amines by asymmetric reduction of prochiral ketones with coenzyme and establish a coupled double enzyme reaction system with formate dehydrogenase( FDH).The results of enzyme activity detection and high performance liquid chromatography show that the optimal reaction condition is that the concentration ratio of Am DH to FDH is 4 ∶ 1 and the concentration of substrate and the concentration of coenzyme are 10 mmol/L and 0. 025 mmol/L,respectively. In order to improve the stability of the system,the polyethylenimine( PEI) coating on the enzyme surface to form polyethyleneimine-diphenylase complex as the template and catalyst is employed for the condensation of Ti-BALDH to form immobilized double enzyme coupling system. Compared with the free double enzyme system,the coenzyme can be recycled and regenerated efficiently in the microenvironment of the immobilized enzyme system. It greatly improves the catalytic efficiency of the multi-enzyme coupling system.
引文
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[16]LIN Peng,ZHANG Yonghui,REN Hong,et al. Assembly of graphene oxide-formate dehydrogenase composites by nickel-coordination with enhanced stability and reusability[J]. Engineering in Life Sciences,2018,18(5):326-333.
[17]ANDERSSON M M,BRECCIA J D,HATTI-KAUL R.Stabilizing effect of chemical additives against oxidation of lactate dehydrogenase[J]. Biotechnology and Applied Biochemistry,2000,32(3):145-153.
[2]KASPRZYK H B. Pharmacologically active compounds in the environment and their chirality[J]. Chemical Society Reviews,2010,39(11):4466.
[3]BROOKS W H,GUIDA W C,DANIEL K G. The significance of chirality in drug design and development[J].Current Topics in Medicinal Chemistry,2011,11(7):760-770.
[4]LEISCH H,MORLEY K,LAU P C. Baeyer-Villiger monooxygenases:more than just green chemistry[J].Chemical Reviews,2011,111(7):4165-4222.
[5]DUNN P J. The importance of green chemistry in process research and development[J]. Chemical Society Reviews,2012,41(4):1452-1461.
[6]CHEN Feifei,ZHENG Gaowei,LIU Lei,et al. Reshaping the active pocket of amine dehydrogenases for asymmetric synthesis of bulky aliphatic amines[J]. ACS Catalysis,2018,8(3):2622-2628.
[7]YE Lijuan,TOH H H,YANG Yi,et al. Engineering of amine dehydrogenase for asymmetric reductive amination of ketone by evolving rhodococcus phenylalanine dehydrogenase[J]. ACS Catalysis,2015,5(2):1119-1122.
[8]CORADIN T,LIVAGE J. Effect of some amino acids and peptides on silicic acid polymerization[J]. Colloids and Surfaces B:Biointerfaces,2001,21(4):329-336.
[9]SUMEREL J L,YANG Wenjun,KISAILUS D,et al.Biocatalytically templated synthesis of titanium dioxide[J]. Chemistry of Materials,2003,15(25):4804-4809.
[10]JIANG Yanjun,YANG Dong,ZHANG Lei,et al. Preparation of protamine-titania microcapsules through synergy between layer-by-layer assembly and biomimetic mineralization[J]. Advanced Functional Materials,2009,19(1):150-156.
[11]JIANG Yanjun,SUN Qianyun,JIANG Zhongyi,et al.The improved stability of enzyme encapsulated in biomimetic titania particles[J]. Materials Science&Engineering C,2009,29(1):328-334.
[12]JIANG Yanjun, YANG Dong, ZHANG Lei, et al.Biomimetics synthesis of titania nanoparticles induced by protamine[J]. Dalton Transactions,2008(31):4165-4171.
[13]YUAN Jianjun,JIN Renhua. Bioinspired synthesis of continuous titania coat with tunable nanofiber-based network structure on linear polyethylenimine-covered substrates[J]. Langmuir,2010,26(6):4212-4218.
[14]SEWELL S L,WRIGHT D W. Biomimetic synthesis of titanium dioxide utilizing the R5 peptide derived from Cylindrotheca fusiformis[J]. Chemistry of Materials,2006,18(13):3108-3113.
[15]REN Hong,ZHANG Yonghui,SU Jieru,et al. Encapsulation of amine dehydrogenase in hybrid titania nanoparticles by polyethylenimine coating and templated biomineralization[J]. Journal of Biotechnology,2017,241:33-41.
[16]LIN Peng,ZHANG Yonghui,REN Hong,et al. Assembly of graphene oxide-formate dehydrogenase composites by nickel-coordination with enhanced stability and reusability[J]. Engineering in Life Sciences,2018,18(5):326-333.
[17]ANDERSSON M M,BRECCIA J D,HATTI-KAUL R.Stabilizing effect of chemical additives against oxidation of lactate dehydrogenase[J]. Biotechnology and Applied Biochemistry,2000,32(3):145-153.