Enhanced enzymatic activity and stability by in situ entrapment of α-Glucosidase within super porous p(HEMA) cryogels during synthesis

Sahin Demirci; Mehtap Sahiner; Selehattin Yilmaz; Erdener Karadag; Nurettin Sahiner

doi:10.1016/j.btre.2020.e00534

Enhanced enzymatic activity and stability by in situ entrapment of α-Glucosidase within super porous p(HEMA) cryogels during synthesis

Biotechnol Rep (Amst). 2020 Sep 28:28:e00534. doi: 10.1016/j.btre.2020.e00534. eCollection 2020 Dec.

Authors

Sahin Demirci^{1

2}, Mehtap Sahiner³, Selehattin Yilmaz¹, Erdener Karadag⁴, Nurettin Sahiner^{1

2

5

6}

Affiliations

¹ Department of Chemistry, Faculty of Sciences and Arts, Canakkale Onsekiz Mart University Terzioglu Campus, 17100, Canakkale, Turkey.
² Nanoscience and Technology Research and Application Center, Canakkale Onsekiz Mart University Terzioglu Campus, 17100, Canakkale, Turkey.
³ Department of Fashion Design, Faculty of Canakkale Applied Science, Canakkale Onsekiz Mart University Terzioglu Campus, 17100, Canakkale, Turkey.
⁴ Department of Chemistry, Faculty of Sciences and Arts, Aydın Adnan Menderes University, 09010, Aydın, Turkey.
⁵ Department of Chemical and Biomolecular Engineering, University of South Florida, Tampa, FL, 33620 USA.
⁶ Department of Ophthalmology, Morsani College of Medicine, University of South Florida, 12901 Bruce B Downs B. Downs Blv., MDC 21, Tampa, FL 33612, USA.

Abstract

Here, poly(2-hydroxyethyl methacrylate) (p(HEMA)) cryogel were prepared in the presence 0.48, 0.96, and 1.92 mL of α-Glucosidase enzyme (0.06 Units/mL) solutions to obtain enzyme entrapped superporous p(HEMA) cryogels, donated as α-Glucosidase@p(HEMA)-1, α-Glucosidase@p(HEMA)-2, and α-Glucosidase@p(HEMA)-3, respectively. The enzyme entrapped p(HEMA) cryogels revealed no interruption for hemolysis and coagulation of blood rendering viable biomedical application in blood contacting applications. The α-Glucosidase@p(HEMA)-1 was found to preserve its' activity% 92.3 ± 1.4 % and higher activity% against free α-Glucosidase enzymes in 15-60℃ temperature, and 4-9 pH range. The K_m and V_max values of α-Glucosidase@p(HEMA)-1 cryogel was calculated as 3.22 mM, and 0.0048 mM/min, respectively versus 1.97 mM, and 0.0032 mM/min, for free enzymes. The α-Glucosidase@p(HEMA)-1 cryogel was found to maintained enzymatic activity more than 50 % after 10 consecutive uses, and also preserved their activity more than 50 % after 10 days of storage at 25 ℃, whereas free α-Glucosidase enzyme maintained only 1.9 ± 0.9 % activity under the same conditions.

Keywords: Enzymatic reaction; Enzyme; Enzyme immobilization/entrapment; Super porous cryogel; α-Glucosidase.