A novel alanine dehydrogenase (AlaDH; EC.1.4.1.1) was isolated from Amycolatopsis sulphurea and the AlaDH gene was cloned into a pET28a(+) plasmid and expressed in E. coli BL21 (DE3). The molecular mass of this enzyme was calculated as 41.09 kDa and the amino acid residues of the pure protein indicated the presence of N terminus polyhistidine tags. Its enzyme kinetic values were K_m 2.03 mM, k_cat 13.24 (s⁻¹), and k_cat/K_m 6.53 (s⁻¹ mM⁻¹). AlaDH catalyzes the reversible conversion of l-alanine and pyruvate, which has an important role in the TCA energy cycle. Maximum AlaDH activity occurred at about pH 10.5 and 25 °C for the oxidative deamination of l-alanine. AlaDH retained about 10% of its relative activity at 55 °C and it remained about 90% active at 50 °C. These findings show that the AsAlaDH from A. sulphurea has the ability to produce valuable molecules for various industrial purposes and could represent a new potential biocatalyst for biotechnological applications after further characterization and improvement of its catalytic properties.

一种新型丙氨酸脱氢酶 (AlaDH; EC.1.4.1.1) 从Amycolatopsis sulfurea 中分离出来，并将 AlaDH 基因克隆到 pET28a(+) 质粒中并在大肠杆菌BL21 (DE3) 中表达。该酶的分子量计算为 41.09 kDa，纯蛋白质的氨基酸残基表明存在 N 端多组氨酸标签。其酶动力学值为K _m 2.03 mM、k _cat 13.24 (s ^-1 )和k _cat /K _m 6.53 (s ^-1  mM ^-1 )。AlaDH 催化l的可逆转化-丙氨酸和丙酮酸，在 TCA 能量循环中具有重要作用。最大 AlaDH 活性发生在约 pH 值 10.5 和 25 °C 时，用于l-丙氨酸的氧化脱氨。AlaDH 在 55 °C 时保持其相对活性的 10% 左右，在 50 °C 时保持约 90% 的活性。这些发现表明，作为从AlaDH A. sulphurea有可能产生有价值的分子各种工业用途的能力，并可能代表了经过进一步的表征和催化性能的改进生物技术应用新的潜在生物催化剂。