Enzyme-Nanoporous Gold Biocomposite: Excellent Biocatalyst with Improved Biocatalytic Performance and Stability

Background: Applications involving biomolecules, such as enzymes, antibodies, and other proteins as well as whole cells, are often hampered by their unstable nature at extremely high temperature and in organic solvents. Methodology/Principal Findings: We constructed enzyme-NPG biocomposites by assembling various enzymes onto the surface of nanoporous gold (NPG), which showed much enhanced biocatalytic performance and stability. Various enzymes with different molecular sizes were successfully tethered onto NPG, and the loadings were 3.6, 3.1 and 0.8 mg g 21 for lipase, catalase and horseradish peroxidase, respectively. The enzyme-NPG biocomposites exhibited remarkable catalytic activities which were fully comparable to those of free enzymes. They also presented enhanced stability, with 74, 78 and 53 % of enzymatic activity retained after 20 successive batch reactions. Moreover, these novel biocomposites possessed significantly enhanced reaction durability under various thermal and in organic solvent systems. In a sample transesterification reaction, a high conversion rate was readily achieved by using the lipase-NPG biocomposite. Conclusion/Significance: These nano-biocomposite materials hold great potential in applications such as biosensing, molecular electronics, catalysis, and controlled delivery

Structures of the high-valent metal-ion haem-oxygen intermediates in peroxidases, oxygenases and catalases

Peroxidases, oxygenases and catalases have similar high-valent metal-ion intermediates in their respective reaction cycles. In this review, haem-based examples will be discussed. The intermediates of the haem-containing enzymes have been extensively studied for many years by different spectroscopic methods like UV-Vis, EPR (electron paramagnetic resonance), resonance Raman, Mossbauer and MCD (magnetic circular dichroism). The first crystal structure of one of these high-valent intermediates was on cytochrome c peroxidase in 1987. Since then, structures have appeared for catalases in 1996, 2002, 2003, putatively for cytochrome P450 in 2000, for myoglobin in 2002, for horseradish peroxidase in 2002 and for cytochrome c peroxidase again in 1994 and 2003. This review will focus on the most recent structural investigations for the different intermediates of these proteins. The structures of these intermediates will also be viewed in light of quantum mechanical (QM) calculations on haem models. In particular quantum refinement, which is a combination of QM calculations and crystallography, will be discussed. Only small structural changes accompany the generation of these intermediates. The crystal structures show that the compound I state, with a so called pi-cation radical on the haem group, has a relatively short iron-oxygen bond (1.67-1.76 A) in agreement with a double-bond character, while the compound 11 state or the compound I state with a radical on an amino acid residue have a relatively long iron-oxygen bond (1.86-1.92 angstrom) in agreement with a single-bond character where the oxygen-atom is protonated