Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Cascavel, Brazil.
Centro Universitário Fundação Assis Gurgacz, Cascavel, Brazil.
J Food Biochem. 2021 Apr;45(4):e13654. doi: 10.1111/jfbc.13654. Epub 2021 Feb 17.
The Cunninghamella echinulata PA3S12MM fungus is a great producer of invertases in a growth medium supplemented by apple peels. The enzyme was purified 4.5 times after two chromatographic processes, and it presented a relative molecular mass of 89.2 kDa. The invertase reached maximum activity at pH of 6 and at 60°C, in addition to presenting stability in alkaline pH and thermal activation at 50°C. The enzymatic activity increased in the presence of Mn and dithiothreitol (DTT), while Cu and Z ions inhibited it. Also, DTT showed to protect enzymatic activity. The apparent values for K , V , and K for the sucrose hydrolysis were, respectively, 173.8 mmol/L, 908.7 mmol/L min , and 1,388.79 s . The carbohydrate content was of 83.13%. The invertase presented hydrolytic activity over different types of glycosidic bonds, such as α1 ↔ 2β (sucrose), α1 → 4 (polygalacturonic acid), α1 → 4 and α1 → 2 (pectin), and α1 ↔ 1 (trehalose), indicating that the enzyme is multifunctional. Thus, the biochemical properties showed by the C. echinulata PA3S12MM suggest a broad industrial application, such as in the biomass hydrolysis or in the food industry. PRACTICAL APPLICATIONS: Invertases are hydrolytic enzymes employed in several industrial sectors. Given their great importance for the economy and several industrial sectors, there is a growing interest in microorganisms producing this enzyme. The analysis of the biochemical properties of invertase in C. echinulata PA3S12MM suggest applications in the food industry. Due to its increased hydrolytic activity, the hydrolysis process of the sucrose may employ invertase for the production of invert sugar. The stability at alkaline pH suggests an application in the development of enzymatic electrodes for the quantification of sucrose in food and beverage. The multifunctional activity may work in the biomass hydrolysis or saccharification of by-products for the extraction of fermentable sugars. The high level of invertase N-linked glycosylation of invertase grants this enzyme thermal stability at high temperatures, in addition to resistance against the action of proteases, which are desirable characteristics for the application of this enzyme in industrial processes.
盾状卷枝毛霉 PA3S12MM 真菌是一种在添加苹果皮的生长培养基中大量生产转化酶的真菌。经过两次色谱过程，该酶被纯化了 4.5 倍，其相对分子质量为 89.2 kDa。该转化酶在 pH 为 6 和 60°C 时达到最大活性，此外在碱性 pH 下稳定，在 50°C 时热激活。在 Mn 和 DTT（二硫苏糖醇）存在下，酶活性增加，而 Cu 和 Z 离子抑制其活性。此外，DTT 显示出保护酶活性的作用。蔗糖水解的表观 K 、 V 和 K 值分别为 173.8 mmol/L、908.7 mmol/L min 和 1.388.79 s。碳水化合物含量为 83.13%。该转化酶对不同类型的糖苷键具有水解活性，如α1↔2β（蔗糖）、α1→4（聚半乳糖醛酸）、α1→4 和α1→2（果胶）以及α1↔1（海藻糖），表明该酶具有多功能性。因此，盾状卷枝毛霉 PA3S12MM 的生化特性表明其具有广泛的工业应用，例如在生物质水解或食品工业中。实际应用：转化酶是在几个工业领域中使用的水解酶。鉴于它们对经济和几个工业部门的重要性，人们对生产这种酶的微生物越来越感兴趣。对 C. echinulata PA3S12MM 中转化酶的生化特性进行分析表明，其在食品工业中有应用潜力。由于其水解活性增加，蔗糖的水解过程可能会使用转化酶生产转化糖。在碱性 pH 下的稳定性表明，它可用于开发用于定量食品和饮料中蔗糖的酶电极。多功能活性可用于生物质水解或副产品的糖化，以提取可发酵糖。转化酶 N 连接糖基化的高水平赋予了该酶在高温下的热稳定性，以及对蛋白酶作用的抗性，这是该酶在工业过程中应用的理想特性。
