Enzymatic degradation of ginkgolic acid by laccase immobilized on novel electrospun nanofiber mat

Abstract

BACKGROUND Ginkgo biloba leaf extract contains many active ingredients that are beneficial for health. However, ginkgolic acid, one of the major components found in G. biloba extract, may cause serious allergic and toxic side effects. The purpose of this study is to immobilize the laccase system on the electrospun nylon fiber mat (NFM) to hydrolyze the ginkgolic acid in G. biloba leaf extract efficiently. RESULTS Novel electrospinning technology successfully produced high‐quality nanoscopic fiber mats made of a mixture of multi‐walled carbon nanotube and nylon 6,6. Laccase that was immobilized onto the NFM exhibited much higher efficiency in the catalyzation of 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt (ABTS) than nylon 6,6 pellets. After being immobilized onto the NFM, the pH and temperature stability of laccase were significantly improved. The NFM‐immobilized laccase could maintain more than 50% of its original activity even after 40 days of storage or 10 operational cycles. The kinetic parameters, including rate constant (K), the time (τ50) in which 50% of ginkgolic acid hydrolysis was reached, the time (τcomplete) required to achieve complete ginkgolic acid hydrolysis, Km and Vmax were determined, and were 0.07 ± 0.01 min−1, 8.97 ± 0.55 min, 45.45 ± 2.79 min, 0.51 ± 0.09 mM and 0.49 ± 0.03 mM min−1 mg−1, respectively. CONCLUSION The result successfully demonstrated the strong potential of using novel electrospun nanofiber mats as enzyme immobilization platforms, which could significantly enhance enzyme activity and stability. © 2020 Society of Chemical Industry

Current Status and Future Perspectives of Supports and Protocols for Enzyme Immobilization

“…High voltage is applied to a polymer solution and the sample collector to produce electric field jets supporting the formation of fibers through solvent evaporation during the process [222,223]. The use of the technique focuses on the main discussions intrinsically connected with the area; it is interpreted as an innovative, effective technique with low cost and versatility that can be applied in several industrial fields [220,[224][225][226]. The electrified nanofibers have attracted the attention of enzyme engineering and biocatalysis, being considered a potential tool because of their numerous advantages: high surface area, multiple fixation points to the support, high porosity, interconnectivity, high thermal resistance, pH stability, and several solvents [216,220,222,223,227].…”

“…Figure 5 shows how the process of enzyme immobilization by electrospinning occurs. Several studies have reported different proposals to immobilize enzymes in electrophile nanofibers [216,219,[223][224][225][226][227][228]. These proposals mainly include binding the enzyme on the surface of nanofibers and enzyme encapsulation [216,219,223,228].…”

Hydrophilic Nonwoven Nanofiber Membranes as Nanostructured Supports for Enzyme Immobilization

“… 57 − 64 Laccases have attracted proven interest in biosensing and bioremediation, and they receive increasing attention in biocatalytic synthesis. 57 , 59 , 60 , 62 − 64 Galactose oxidase finds application in biosensing, production of carbohydrate-based reactive aldehydes, and valorization of hydroxymethylfurfural. 58 , 64 These preliminary results are a promising basis for the application of nonwoven membranes in enzyme technology.…”

Fabrication and characterization of fast dissolving glycerol monolaurate microemulsion encapsulated gelatin nanofibers with antimicrobial activity

“…Electrospinning is an effective and versatile method of fabrication from polymer solutions or melted polymer continuous fibers with a diameter ranging from nanometers to micrometers. 1 As a result of a tunable diameter, high porosity and a high specific surface area, electrospun fiber is a suitable candidate for the targeted delivery of labile and sensitive compounds, 2 bioactive encapsulation, 3,4 enzyme immobilization, 5 and nano-sensor 6 and active packaging 7 applications. In recent years, interest in the use of biopolymer electrospun nanofibers for various food applications has increased because of environmental and safety concerns.…”