Hollow Mesoporous Carbon-Based Enzyme Nanoreactor for the Confined and Interfacial Biocatalytic Synthesis of Phytosterol Esters

Abstract

Rationally designing carriers to obtain efficient and stable immobilized enzymes for the production of food raw materials is always a challenge. In this work, hollow cube carbon (HMC) as a carrier of Candida rugosa lipase (CRL) was prepared to construct a Pickering interfacial biocatalysis system, which was applied to biphasic biocatalysis. For comparison, the nonporous carbon (HC) and porous MoS 2 (HMoS 2 ) were also designed. On these grounds, p-NPP and linolenic acid were selected as the representative substrates for hydrolysis and esterification reactions. Under the optimal conditions, the protein loading amount, specific activity, and expressed activity of CRL immobilized on HMC (HMC@CRL) were 167.2 mg g −1 , 5.41 U mg −1 , and 32.34 U/ mg protein, respectively. In the "oil−water" biphase, the relative hydrolytic activity of HMC@CRL was higher than that of HC@ CRL, HMoS 2 @CRL, and CRL by 50, 68, and 80%, respectively, as well as itself in one phase. Compared to other reports (1.13%), HMC@CRL demonstrated a satisfactory hydrolysis rate (3.02%) and was the fastest among all other biocatalysts in the biphase. Moreover, compared with the free CRL in one-phase system, the Pickering interfacial biphasic biocatalyst, HMC@CRL, exhibited a higher esterification rate (85%, 2.7-fold enhancement). Therefore, the HMC@CRL nanoreactors had more optimal performance in the field of biomanufacturing and food industry.

Octadecyl and sulfonyl modification of diatomite synergistically improved the immobilization efficiency of lipase and its application in the synthesis of pine sterol esters

“…The decrease in esterification yield is most likely due to the unavoidable loss of the immobilized enzyme during the separation from the reaction system, as well as denaturation and inactivation of the enzyme during the reaction. For instance, Zhang et al suggested that only 82.0% activity of CRL immobilized with a hollow cubic carbon can be maintained in the second cycle [67]. However, in this study, our novel-designed OSMD immobilization can maintain the residual activity as high as 90.6% in two cycles, which suggests that the novel-designed OSMD in this study is a good candidate in lipase immobilization and has great potential for application in further food biomanufacturing.…”

Octadecyl and sulfonyl modification of diatomite synergistically improved the immobilization efficiency of lipase and its application in the synthesis of pine sterol esters

“…suggested that only 82.0% activity of CRL immobilized with a hollow cubic carbon can be maintained in the second cycle. [ 75 ] However, in this study, our novel‐designed OSMD immobilization can maintain the residual activity as high as 91.5% in two cycles, which suggests that the novel‐designed OSMD in this study is a good candidate in lipase immobilization and has great potential for application in further food biomanufacturing.…”

Lipase-Entrapped Polydopamine (PDA) Nanobottles for Near-Infrared (NIR)-Triggered Pickering Interfacial Biocatalysis (PIB)

“…Additionally, other studies have also demonstrated a trend of temperature-dependent changes in enzyme activity. 60,61 Figure 7c also shows that PDA@G-CRL has greater thermal stability in comparison to free CRL. The enzyme was encased in gelatin, and the movement of the embedded carrier was limited, and the requirement for enhanced reaction substrate diffusion at higher temperatures might all contribute to these results.…”