Abstract

The L-asparaginase (ASPN) enzyme has received recognition in various applications including acrylamide degradation in the food industry. The synthesis and application of thermostable ASPN enzymes is required for its use in the food sector, where thermostable enzymes can withstand high temperatures. To achieve this goal, the bacterium Bacillus subtilis was isolated from the hot springs of Tapovan for screening the production of thermostable ASPN enzyme. Thus, ASPN with a maximal specific enzymatic activity of 0.896 U/mg and a molecular weight of 66 kDa was produced from the isolated bacteria. The kinetic study of the enzyme yielded a Km value of 1.579 mM and a Vmax of 5.009 µM/min with thermostability up to 100 min at 75 °C. This may have had a positive indication for employing the enzyme to stop polyacrylamide from being produced. The current study has also been extended to investigate the interaction of native and mutated ASPN enzymes with acrylamide. This concluded that the M₁₀ (with 10 mutations) has the highest protein and thermal stability compared to the wild-type ASPN protein sequence. Therefore, in comparison to a normal ASPN and all other mutant ASPNs, M₁₀ is the most favorable mutation. This research has also demonstrated the usage of ASPN in food industrial applications.

Graphical Abstract

中文翻译：

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用于聚丙烯酰胺抑制和计算机突变分析的耐热细菌 L-天冬酰胺酶

摘要

L-天冬酰胺酶 (ASPN) 已在多种应用中得到认可，包括食品工业中的丙烯酰胺降解。热稳定性ASPN酶的合成和应用是其在食品领域的应用所必需的，因为热稳定性酶可以承受高温。为了实现这一目标，从Tapovan的温泉中分离出枯草芽孢杆菌，用于筛选生产耐热ASPN酶。因此，从分离的细菌中产生了最大比酶活性为0.896 U/mg、分子量为66 kDa的ASPN。该酶的动力学研究得出 Km 值为 1.579 mM，Vmax 为 5.009 µM/min，在 75 °C 下热稳定性长达 100 分钟。这可能对使用酶来阻止聚丙烯酰胺的产生具有积极的意义。目前的研究还扩展到研究天然和突变的 ASPN 酶与丙烯酰胺的相互作用。由此得出结论，与野生型 ASPN 蛋白质序列相比，M ₁₀ （具有 10 个突变）具有最高的蛋白质和热稳定性。因此，与正常 ASPN 和所有其他突变 ASPN 相比，M ₁₀是最有利的突变。这项研究还展示了 ASPN 在食品工业应用中的用途。

图形概要