A novel amidase (TAM) was identified and cloned from the genome of Thauera sinica K11. The recombinant protein was purified to homogeneity by one-step affinity chromatography for up to 26.4-fold with a yield of 38.1%. Gel filtration chromatography and SDS-PAGE revealed that the enzyme was a tetramer with a subunit of approximately 37.5 kDa. The amidase exhibited the maximum acyl transfer activity at 45 °C and pH 7.0, and it was highly stable over a wide pH range of 6.0–11.0. Inhibition of enzyme activity was observed in the presence of metal ions, thiol reagents and organic solvents. TAM showed a broad substrate spectrum toward aliphatic, aromatic and heterocyclic amides. For linear aliphatic monoamides, the acyl transfer activity of TAM was decreased with the extension of the carbon chain length, and thus the highest activity of 228.2 U/mg was obtained when formamide was used as substrate. This distinct selectivity of amidase to linear aliphatic monoamides expanded the findings of signature amidases to substrate specificity.

鉴定了一种新型的酰胺酶（TAM），并从Thauera sinica的基因组中进行了克隆。K11。通过一步亲和层析将重组蛋白纯化至同质，最多纯化26.4倍，收率为38.1％。凝胶过滤色谱和SDS-PAGE表明该酶是四聚体，其亚基约为37.5kDa。酰胺酶在45°C和pH 7.0时表现出最大的酰基转移活性，并且在6.0-11.0的宽pH范围内都非常稳定。在金属离子，硫醇试剂和有机溶剂的存在下观察到酶活性的抑制。TAM对脂族，芳族和杂环酰胺显示出较宽的底物光谱。对于直链脂族单酰胺，TAM的酰基转移活性随碳链长度的延长而降低，因此当使用甲酰胺作为底物时，最高活性为228.2 U / mg。