Demirci Sahin, Sahiner Mehtap, Yilmaz Selehattin, Karadag Erdener, Sahiner Nurettin
Department of Chemistry, Faculty of Sciences and Arts, Canakkale Onsekiz Mart University Terzioglu Campus, 17100, Canakkale, Turkey.
Nanoscience and Technology Research and Application Center, Canakkale Onsekiz Mart University Terzioglu Campus, 17100, Canakkale, Turkey.
Biotechnol Rep (Amst). 2020 Sep 28;28:e00534. doi: 10.1016/j.btre.2020.e00534. eCollection 2020 Dec.
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 and V 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.
在此,在0.48、0.96和1.92 mL的α-葡萄糖苷酶(0.06单位/mL)溶液存在下制备聚(甲基丙烯酸2-羟乙酯)(p(HEMA))冷冻凝胶,以获得包埋酶的超大孔p(HEMA)冷冻凝胶,分别命名为α-葡萄糖苷酶@p(HEMA)-1、α-葡萄糖苷酶@p(HEMA)-2和α-葡萄糖苷酶@p(HEMA)-3。包埋酶的p(HEMA)冷冻凝胶在血液溶血和凝血方面无干扰,在血液接触应用中具有可行的生物医学应用。发现α-葡萄糖苷酶@p(HEMA)-1在15 - 60℃温度和4 - 9的pH范围内,相对于游离α-葡萄糖苷酶,其活性保留率为92.3±1.4%,且活性更高。α-葡萄糖苷酶@p(HEMA)-1冷冻凝胶的K值和V值分别计算为3.22 mM和0.0048 mM/min,而游离酶的K值和V值分别为1.97 mM和0.0032 mM/min。发现α-葡萄糖苷酶@p(HEMA)-1冷冻凝胶在连续使用10次后仍保持超过50%的酶活性,并且在25℃储存10天后其活性也保留超过50%,而在相同条件下,游离α-葡萄糖苷酶仅保持1.9±0.9%的活性。