Gelatin-Immobilized Manganese Peroxidase with Novel Catalytic Characteristics and Its Industrial Exploitation for Fruit Juice Clarification Purposes
Gelatin-Immobilized Manganese Peroxidase with Novel Catalytic Characteristics and Its Industrial Exploitation for Fruit Juice Clarification Purposes
复制标题
DOI：
10.1007/s10562-016-1848-9
复制
发表时间：
2016-11-01
影响因子：
2.8
通讯作者：
Zhang, Xuehong
中科院分区：
文献类型：
--
作者：
Bilal, Muhammad;Asgher, Muhammad;Zhang, Xuehong
本研究以戊二醛(GLA)活化的明胶水凝胶为载体，在优化前的生长环境中对灵芝IBL-05产生的锰过氧化物酶(MNP；E.C.1.11.1.13)进行了包埋。在明胶辅助固定化条件下，明胶的最佳固定化率为83.2+/-2.91%，当酶浓度为0.6 mg/m L，活化时间为2 h时，固定化率为20.0%(w/v)，GLA为0.25%(v/v)。与水相相比，不溶性MNP在pH 6.0和60A℃时活性最高。酶固定化显著提高了室内分离的MNP的热稳定性。在60℃时，游离MNP的最大活性下降到14.2A+/-1.4%，而固定化MNP在120min后保持了70.18A+/-3.2%的活性。为了探索固定化MNP的工业适用性，在填充床反应器系统中对固定化MNP的苹果和橙汁澄清特性进行了测试。固定化MNP对苹果原汁的脱苦效果良好，原汁色泽降低42.7%，浑浊度降低36.3%。橙汁的降色率和浑浊度分别为51.5%和43.6%。连续6次循环后，固定化MNP仍能保持其初始活性的60.0%以上。总体而言，凝胶包裹的MNP的催化、热稳定性和透明度改善特性表明，酶处理在生物技术开发中具有很高的潜力。
In the present study, glutaraldehyde (GLA) activated gelatin hydrogel was used as a solid support to encapsulate the manganese peroxidase (MnP; E.C. 1.11.1.13) produced by Ganoderma lucidum IBL-05 under pre-optimized growth environment. Through gelatin-assisted immobilization, a maximal of 83.2 +/- 2.91 % immobilization yield was achieved at optimum conditions of gelatin; 20.0 % (w/v), GLA 0.25 % (v/v) after 2 h activation time using 0.6 mg/mL of enzyme concentration. In contrast to aqueous form, the insolubilized MnP presented its maximum activity at pH 6.0 and 60 A degrees C. Inevitably, enzyme immobilization significantly (P < 0.05) increased the thermal stability profile of in-house isolated MnP. At 60 A degrees C, maximum activity of free MnP decreased to 14.2 A +/- 1.4 %, whereas immobilized MnP retained 70.18 A +/- 3.2 % of its original activity after 120 min. To explore the industrial applicability of MnP, the immobilized MnP was tested for apple and orange fruit juice clarification features in a packed bed reactor system. The immobilized MnP showed commendable results in the de-bittering's of investigated fruit juices, decreasing 42.7 % of the original apple juice color and 36.3 of its turbidity. Whereas, the color and turbidity reduction characteristics of orange juice were 51.5 and 43.6 %, respectively. After six consecutive cycles, the immobilized-MnP was able to retain more than 60.0 % of its initial activity. Collectively, catalytic, thermo-stability and clarity amelioration features of the gel-entrapped MnP suggest a high potential of enzymatic treatment for biotechnological exploitability.