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Enhanced enzymatic activity and stability by in situ entrapment of α-Glucosidase within super porous p(HEMA) cryogels during synthesis
Journal article   Open access   Peer reviewed

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 and Nurettin Sahiner
Biotechnology reports (Amsterdam, Netherlands), Vol.28, p.e00534
12-01-2020
PMID: 33024715

Abstract

Enzymatic reaction Enzyme Enzyme immobilization/entrapment Super porous cryogel α-Glucosidase
•The α-Glucosidase was successfully entrapment into p(HEMA) cryogels at −18 °C.•Higher stability for α-Glucosidase@p(HEMA) than free one at wide range of T and pHs.•∼1.5-fold increase in Vmax value for the entrapped α-Glucosidase was accomplished.•Entrapped enzyme, α-Glucosidase maintained >50 % activity after 10 repeated use.•Entrapped α-Glucosidase can be stored at 25 °C for 10 days with >50 % activity. 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 Km and Vmax 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.
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