A putative diamine oxidase (DAO) from Yarrowia lipolytica PO1f (DAO-1) was homologously recombinantly integrated into the genome of Y. lipolytica PO1f using the CRISPR-Cas9 system for the subsequent DAO production in a bioreactor. Thereby, it was proven that the DAO-1 produced was indeed a functional DAO. The cultivation yielded 2343 ± 98 nkat/L_culture with a specific DAO activity of 1301 ± 54.2 nkat/g_protein, which was a 93-fold increase of specific DAO activity compared to the native Y. lipolytica PO1f DAO-1 production. The DAO-1 showed a broad substrate selectivity with tyramine, histamine, putrescine and cadaverine being the most favored substrates. It was most active at 40 °C, pH 7.2 in Tris-HCl buffer (50 mM) (with histamine as substrate), which is comparable to human and porcine DAOs. The affinity of DAO-1 towards histamine was lower compared to mammalian DAOs (K_m = 2.3 ± 0.2 mM). Nevertheless, DAO-1 degraded around 75% of the histamine used in a bioconversion experiment with a food-relevant concentration of 150 mg/L. With its broad selectivity for the most relevant biogenic amines in foods, DAO-1 from Y. lipolytica PO1f is an interesting enzyme for application in the food industry for the degradation of biogenic amines.

从推定的二胺氧化酶（DAO）解脂耶氏酵母Po1f（DAO-1）同源重组整合到基因组中解脂亚罗威阿酵母使用CRISPR-Cas9系统用于随后DAO生产生物反应器中酵母Po1f。从而证明产生的DAO-1确实是一个功能性的DAO。培养产生 2343 ± 98 nkat/L 的_培养物，具有 1301 ± 54.2 nkat/g_蛋白的特定 DAO 活性，与天然解脂耶氏酵母相比，特定 DAO 活性增加了 93 倍PO1f DAO-1 生产。DAO-1 显示出广泛的底物选择性，其中酪胺、组胺、腐胺和尸胺是最受青睐的底物。它在 40 °C、pH 7.2 的 Tris-HCl 缓冲液 (50 mM)（以组胺为底物）中最活跃，与人和猪 DAO 相当。与哺乳动物 DAO 相比，DAO-1 对组胺的亲和力较低 ( K _m = 2.3 ± 0.2 mM)。尽管如此，DAO-1 降解了大约 75% 的组胺，用于生物转化实验中，与食物相关的浓度为 150 mg/L。由于对食品中最相关的生物胺具有广泛的选择性，来自解脂耶氏酵母PO1f 的DAO-1是一种有趣的酶，可用于食品工业中降解生物胺。