Fast Immobilization of Human Carbonic Anhydrase II on Ni-Based Metal-Organic Framework Nanorods with High Catalytic Performance

Abstract

Carbonic anhydrase (CA) has received considerable attention for its ability to capture carbon dioxide efficiently. This study reports a simple strategy for immobilizing recombinant carbonic anhydrase II from human (hCA II) on Ni-based MOFs (Ni-BTC) nanorods, which was readily achieved in a one-pot immobilization of His-tagged hCA II (His-hCA II). Consequently, His-hCA II from cell lysate could obtain an activity recovery of 99% under optimal conditions. After storing for 10 days, the immobilized His-hCA II maintained 40% activity while the free enzyme lost 91% activity. Furthermore, during the hydrolysis of p-nitrophenyl acetic acid, immobilized His-hCA II exhibited excellent reusability and still retained more than 65% of the original activity after eight cycles. In addition, we also found that Ni-BTC had no fixation effect on proteins without histidine-tag. These results show that the Ni-BTC MOFs have a great potential with high efficiency for and specific binding of immobilized enzymes.

Characterization of Gentisate 1,2-Dioxygenase from Pseudarthrobacter phenanthrenivorans Sphe3 and Its Stabilization by Immobilization on Nickel-Functionalized Magnetic Nanoparticles

“…These results are in agreement with previous studies that reported an increase in the K m value after immobilization [40]. Furthermore, Jiao et al, recently reported an increase in the K m value after immobilizing a recombinant carbonic anhydrase II from human (hCA II) on Ni-based metal-organic framework nanorods [80]. Moreover, the apparent V max value was significantly decreased after immobilization, indicating lower enzymatic reaction rates of the substrate, probably due to the steric hindrance of the support after immobilization.…”

“…This result is in agreement with other studies where immobilization of enzymes extends their storage stability [87,88]. For example, during 10 d of storage, the His-tagged carbonic anhydrase II immobilized on Ni-based metal-organic framework nanorods maintained 40% activity, while the free enzyme lost 91% of its initial activity [80]. These results indicate that the immobilization of His-tagged enzymes on Ni-modified supports could provide stability and mechanical protection towards denaturation against prolonged storage.…”

Direct Biocatalytic Processes for CO2 Capture as a Green Tool to Produce Value-Added Chemicals

“…MOFs are employed in a multitude of applications, with the immobilization of proteins being one of the most promising [ 168 , 169 ]. CA has been encapsulated in different MOFs with different features, most of them being zeolites constituted by imidazolates, with acceptable or excellent results [ 149 , 154 , 170 ]. The enzyme encapsulation within MOFs generates enzyme diffusion through windows that have a smaller size than the cavity.…”

“…Whether or not the term encapsulation can be properly applied to the immobilization of enzymes in MOFs depends on the ratio between the pore and the enzyme size. In any case, an MOF based on Ni(II) showed a high degree of reusability for CA, retaining more than 65% of its activity after eight cycles [ 149 ]. Zinc has also been used as a base for MOFs to immobilize CA.…”

Metal Organic Frameworks for Advanced Applications

“…In particular, the type of oxidant and the way to supply it to the reaction medium influenced the catalytic results. Jiao et al [10] described the fast immobilization of human carbonic anhydrase II on Ni-based MOF nanorods with high catalytic performance. These results showed that the Ni-MOFs have great potential and high efficiency for the specific binding of immobilized enzymes.…”