Lipase is one of the most utilized classes of biocatalysts. It can be widely used in the enzymatic organic synthesis and clinical analysis. Denaturation of lipase, which destroys its catalytic activity and stability, can be induced by heat or organic solvents. Thus, the lipase-NPG biocomposite was investigated under different experimental conditions. The results showed that the novel biocomposite possessed significantly enhanced reaction durability under various thermal and in organic solvent systems. In addition, a conservative estimation suggested that the lipase-NPG biocomposite could retain its high activity for at least 240 h in transesterification reaction system. These results were markedly better than previous reports where a significant decrease in activity was observed within 10 recycles in biodiesel production using immobilized lipase. The high conversion rate once again confirmed that NPG is an excellent support for enzyme immobilization. The above results clearly suggested that the immobilization using bio-compatible and highly conductive NPG could enhance the stability of enzymes. The excellent catalytic performance and stability of the enzyme-NPG biocomposites might be explained by their physical confinement inside the relatively small pores. Especially, the size match between pore dimension and the molecular diameter of enzymes and the suitability of gold to function as an immobilization medium are of key importance in achieving high enzymatic stability. This involves the adsorption of the enzyme with its active site oriented away from the porous surface with little leaching yet sufficient mobility to retain catalytic activity. Moreover, it is well accepted that the interaction of nanoscale gold with NH2 was as strong as that with the commonly used SH. The lipase from Pseudomonas cepacia has fourteen lysine residues and six cysteine residues. The covalent attachments of enzymes by the amino and mercapto groups onto the surface of NPG could prevent the enzyme leaching, and the curvatures of the porous surfaces could provide an ideal configuration for multipoint covalent attachments to global enzyme molecules, resulting in better operational stability. To prove this, X-ray photoelectron spectroscopy was used to probe the chemical state of the surface for the lipase-NPG biocomposite. In free lipase, there is only a single broad peak at about 163.7 eV for S2p. In contrast, after immobilization on NPG, it splits into two peaks with binding energy in the range from 162.0 to 165.0 eV corresponding to the chemical states of sulfur in Au-S and S-H, respectively. Additionally, the N1s binding energy of nitrogen in protein at 399.6 eV was also observed in the lipase-NPG biocomposite as shown in Figure 7b, while no the signal of N1s was observed on the bare NPG.