芽孢杆菌的热稳定性增加酶α-Amilase subtilis ITBCCB148 Amobilisasi用沸石杨林安

Yandri, Fathaniah Sejati, Tati Suhartati
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引用次数: 0

摘要

采用沸石固定化技术提高枯草芽孢杆菌ITBCCB148中-淀粉酶的热稳定性。因此,首先我们需要生产、分离和纯化酶。酶的纯化步骤为:硫酸铵分馏、透析、cm -纤维素阳离子交换柱层析。纯化后的酶用沸石固定化。通过比较固定化前后酶的热稳定性来评价固定化成功与否。采用Mandels法和Fuwa法测定α-淀粉酶活性。蛋白质含量采用Lowry法测定。结果表明,纯化后的酶比活性为2473.7 U / mg,比酶粗提物的1285.9 U / mg提高了19倍。纯化酶的最适温度为65℃,固定化酶的最适温度为75℃。纯化酶在65℃下热稳定性测试100分钟表明纯化酶的残留活性为20%;t1 / 2 = 30min, ki = 0.023 min -1, ΔGi = 103.65 kJ mol -1。固定化酶在65℃下热稳定性试验表明,固定化酶残留活性为40%;t1 / 2 = 49 min, ki = 0.014 min -1, ΔGi = 105.03 kJ mol -1。沸石固定化后酶的热稳定性比纯化后的酶提高了1.64倍,表现为ki值降低,半衰期增加,变性能变化增大(ΔGi)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Peningkatan Stabilitas Termal Enzim α-Amilase dari Bacillus subtilis ITBCCB148 dengan Amobilisasi Menggunakan Zeolit
The objective of the research is to increase the thermal stability of -amylase from Bacillus subtilis ITBCCB148 by immobilization using zeolite. For that reason, firstly we need to produce, isolate, and purify the enzyme. The purification of the enzyme was conducted by the following steps: fractionation with ammonium sulphate, dialysis, and CM-cellulose cation exchange column chromatography. The purified enzyme was immobilized using zeolite. The success in immobilization of the enzyme was evaluated by comparing the thermal stability of the enzyme before and after immobilization. Activity of α-amylase was determined by the Mandels and Fuwa method. The protein content was determined based on the method by Lowry. The results showed that the specific activity of purified enzyme was 2473.7 U / mg, increased 19 times compared to crude extract of enzyme having specific activity of 1285.9 U / mg. The purified enzyme has the optimum temperature at 65ºC, while the immobilized enzyme has the optimum temperature at 75ºC. The thermal stability test of the purified enzyme at 65ºC for 100 minutes showed the purified enzyme having residual activity of 20%; t 1 / 2 = 30 min, k i = 0.023 min -1 and ΔGi = 103.65 kJ mol -1 . The thermal stability test of the immobilized enzyme at 65ºC for 100 minutes showed that the immobilized enzyme had residual activity of 40%; t 1/ 2 = 49 min, k i = 0.014 min -1 and ΔGi = 105.03 kJ mol -1 . Immobilization using zeolite has succeeded in increasing the thermal stability of enzyme by 1.64 times compared to the purified enzyme, which is indicated by the decreasing of k i value, the increase of half-life and denaturation energy change (ΔGi).
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