State Key Laboratory of Tea Plant Biology and Utilization, School of Tea and Food Science Technology, Anhui Agricultural University, Hefei, Anhui 230036, People's Republic of China.
School of Life Science, Anhui Agricultural University, Hefei, Anhui 230036, People's Republic of China.
J Agric Food Chem. 2024 Jul 24;72(29):16403-16411. doi: 10.1021/acs.jafc.4c03804. Epub 2024 Jul 14.
As a highly toxic mycotoxin, ochratoxin A (OTA) is widely contaminating agricultural products and has various toxicological effects. Bioenzymes for OTA degradation have shown promising potential for detoxification. Other than the efficient amidohydrolase ADH3 previously, two novel amidohydrolases ADH1 and AMD3 were obtained in this study. During expression, the expressed protein solubility was very low and will limit future industrial application. Here, high copy number integrations were screened, and the amidohydrolases were efficiently secretory expressed by GS115. The protein yields from 1.0 L of fermentation supernatant were 53.5 mg for ADH1, 89.15 mg for ADH3, and 79.5 mg for AMD3. The catalytic efficiency (/) of secretory proteins was 124.95 s mM for ADH3, 123.21 s mM for ADH1, and 371.99 s mM for AMD3. In comparison to expression, the active protein yields substantially increased 15.78-51.53 times. Meanwhile, two novel amidohydrolases (ADH1 and AMD3) showed much higher activity than ADH3 that produced by secretory expression.
作为一种高毒性的真菌毒素，赭曲霉毒素 A（OTA）广泛污染农产品，并具有多种毒理学效应。OTA 降解的生物酶显示出解毒的有前景的潜力。除了之前高效的酰胺水解酶 ADH3 外，本研究还获得了两种新型的酰胺水解酶 ADH1 和 AMD3。在表达过程中，表达的蛋白溶解度非常低，这将限制未来的工业应用。在这里，筛选了高拷贝数的整合，并通过 GS115 有效地分泌表达了酰胺水解酶。从 1.0 L 发酵上清液中获得的蛋白产量分别为 ADH1 53.5mg、ADH3 89.15mg 和 AMD3 79.5mg。分泌蛋白的催化效率（/）分别为 ADH3 124.95s mM、ADH1 123.21s mM 和 AMD3 371.99s mM。与表达相比，活性蛋白产量增加了 15.78-51.53 倍。同时，两种新型的酰胺水解酶（ADH1 和 AMD3）的活性明显高于通过分泌表达产生的 ADH3。
