Cascade Biocatalysis by Multienzyme鈥揘anoparticle Assemblies

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• 作者：Wei Kang ; Jiahui Liu ; Jianpeng Wang ; Yunyu Nie ; Zhihong Guo ; Jiang Xia
• 刊名：Bioconjugate Chemistry
• 出版年：2014
• 出版时间：August 20, 2014
• 年：2014
• 卷：25
• 期：8
• 页码：1387-1394
• 全文大小：464K
• ISSN：1520-4812

文摘

Multienzyme complexes are of paramount importance in biosynthesis in cells. Yet, how sequential enzymes of cascade catalytic reactions synergize their activities through spatial organization remains elusive. Recent development of site-specific protein鈥搉anoparticle conjugation techniques enables us to construct multienzyme assemblies using nanoparticles as the template. Sequential enzymes in menaquinone biosynthetic pathway were conjugated to CdSe-ZnS quantum dots (QDs, a nanosized particulate material) through metal-affinity driven self-assembly. The assemblies were characterized by electrophoretic methods, the catalytic activities were monitored by reverse-phase chromatography, and the composition of the multienzyme鈥換D assemblies was optimized through a progressive approach to achieve highly efficient catalytic conversion. Shorter enzyme鈥揺nzyme distance was discovered to facilitate intermediate transfer, and a fine control on the stoichiometric ratio of the assembly was found to be critical for the maximal synergy between the enzymes. Multienzyme鈥換D assemblies thereby provide an effective model to scrutinize the synergy of cascade enzymes in multienzyme complexes.