Electrospun polyvinyl alcohol/bovine serum albumin biocomposite membranes for horseradish peroxidase immobilization

被引:27
作者
Fazel, Ramin [1 ]
Torabi, Seyed-Fakhreddin [1 ,5 ]
Naseri-Nosar, Pooya [1 ,6 ]
Ghasempur, Salehe [1 ]
Ranaei-Siadat, Seyed-Omid [2 ,3 ]
Khajeh, Khosro [1 ,4 ]
机构
[1] Univ Tehran, Dept Biotechnol, Coll Sci, Tehran, Iran
[2] Shahid Beheshti Univ, Fac Engn & New Technol, Nanobiotechnol Engn Lab, GC, Tehran, Iran
[3] Shahid Beheshti Univ, Prot Res Ctr, Prot Engn Lab, GC, Tehran, Iran
[4] Tarbiat Modares Univ, Fac Biol Sci, Dept Biochem, Tehran, Iran
[5] Yale Univ, Howard Hughes Med Inst, Dept Mol Biophys & Biochem, New Haven, CT 06511 USA
[6] Princeton Univ, Dept Mol Biol, Princeton, NJ 08544 USA
关键词
Biocomposite nanofibrous membranes; Horseradish peroxidase; Immobilization; Electrospinning; Cross-linking; Bovine serum albumin; ENZYME IMMOBILIZATION; POLYMER NANOFIBERS; CROSS-LINKING; FIBROUS MEMBRANES; ADSORPTION; GLUTARALDEHYDE; INACTIVATION; DEGRADATION; PARTICLES; GELATIN;
D O I
10.1016/j.enzmictec.2016.07.002
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Electrospinning, a simple and versatile method to fabricate nanofibrous supports, has attracted attention in the field of enzyme immobilization. Biocomposite nanofibers were fabricated from mixed PVA/BSA solution and the effects of glutaraldehyde treatment, initial BSA concentration and PVA concentration on protein loading were investigated. Glutaraldehyde cross-linking significantly decreased protein release from nanofibers and BSA loading reached as high as 27.3% (w/w). In comparison with the HRP immobilized into the nascent nanofibrous membrane, a significant increase was observed in the activity retention of the enzyme immobilized into the PVA/BSA biocomposite nanofibers. The immobilized HRP was able to tolerate much higher concentrations of hydrogen peroxide than the free enzyme and thus the immobilized enzyme did not demonstrate substrate inhibition. The immobilized HRP retained similar to 50% of the free enzyme activity at 6.4 mM hydrogen peroxide and no significant variation was observed in the KM value of the enzyme for hydrogen peroxide after immobilization. In addition, reusability tests showed that the residual activity of the immobilized HRP were 73% after 11 reuse cycles. Together, these results demonstrate efficient immobilization of HRP into electrospun PVA/BSA biocomposite nanofibers and provide a promising immobilization strategy for biotechnological applications. (C) 2016 Elsevier Inc. All rights reserved.
引用
收藏
页码:1 / 10
页数:10
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