Building a Thermostable Metabolon for Facilitating Coenzyme Transport and In Vitro Hydrogen Production at Elevated Temperature

摘要: To facilitate coenzyme transport and in vitro enzymatic hydrogen production, a multi-enzyme metabolon comprising miniscaffoldin containing three cohesins, dockerin-containing mutant dehydrogenase, diaphorase, Histidine-tagged (His-tagged) NiFe hydrogenase was constructed. As the has very complicated structure cannot be fused directly with dockerin, bifunctional peptide designed. The peptide, in which one terminus contains modified dockerin binding cohesin of other, after chemical modification, binds His-tag hydrogenase, enabled His-tagged proteins to integrated into cohesin-dockerin-based metabolon. exhibited an initial reaction rate 4.5 times that enzyme cocktail at same loading, indicated enhanced However, this unstable owing degradation elevated temperature. Glutaraldehyde used cross-link for locking its spatial organization. cross-linked not only 2.5 times cocktail, but also retained stability 70 °C. amount production catalyzed by nearly twice without glutaraldehyde cross-linking four times 70 °C 22 h reaction.