杂化修饰埃洛石纳米粘土固定内切菊粉酶制备低聚果糖。
Hetero-modification of halloysite nanoclay to immobilize endoinulinase for the preparation of fructooligosaccharides.
机构信息
Carbohydrate and Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala 147 002, Punjab, India; University Centre for Research and Development, Chandigarh University, Gharuan, 140 413 Mohali, Punjab, India.
Carbohydrate and Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala 147 002, Punjab, India.
出版信息
Food Res Int. 2022 Sep;159:111591. doi: 10.1016/j.foodres.2022.111591. Epub 2022 Jun 28.
Present investigation describes immobilization efficiency of endoinulinase onto hetero-functionalized halloysite nanoclay using 3-aminopropyltriethoxysilane and glutaraldehyde as crosslinkers. Under optimal conditions (APTES 0.75%, sonication time 2.25 h, glutaraldehyde 0.75%, activation-time 65 min, immobilized endoinulinase load 60 IU and coupling-time 1 h), maximum yield in enzyme activity (70.65%) and immobilization (89.61%) was obtained. Developed immobilized biocatalyst shown maximum activity at 65 °C and pH 5.0 with wide range thermal (50-80 °C) and pH (4.0-9.0) stability. Increase in half-life (28.70-fold) of immobilized endoinulinase was observed as compared to free enzyme. An enhanced K and reduced V of endoinulinase for inulin was recorded after immobilization. Maximum FOSs production 98.42% was obtained, under optimized conditions (inulin 10%; immobilized endoinulinase load 85 IU; hydrolysis-time 10 h and agitation rate 130 rpm) containing kestose (36.26%), nystose (27.02%), fructofuranosylnystose (9.98%) and FOSs DP 5-9 (25.15%). Developed immobilized biocatalyst exhibited a splendid operational stability for 18 batch cycles.
本研究描述了使用 3-氨丙基三乙氧基硅烷和戊二醛作为交联剂将内切菊粉酶固定在杂化羟硅铝酸盐纳米粘土上的固定化效率。在最佳条件下（APTES 0.75%，超声时间 2.25 h，戊二醛 0.75%，活化时间 65 min，固定化内切菊粉酶负载量 60 IU，偶联时间 1 h），获得了最大的酶活（70.65%）和固定化（89.61%）收率。开发的固定化生物催化剂在 65°C 和 pH 5.0 下表现出最大的活性，具有较宽的热（50-80°C）和 pH（4.0-9.0）稳定性。与游离酶相比，固定化内切菊粉酶的半衰期（28.70 倍）增加。固定化后，内切菊粉酶对菊粉的 K 和 V 降低。在优化条件下（菊粉 10%，固定化内切菊粉酶负载量 85 IU，水解时间 10 h，搅拌速度 130 rpm），获得了 98.42%的最大 FOS 产量，其中包含蔗果三糖（36.26%）、棉子糖（27.02%）、果呋喃糖基棉子糖（9.98%）和 FOSs DP 5-9（25.15%）。开发的固定化生物催化剂在 18 批循环中表现出出色的操作稳定性。
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