Trans-2,3-dihydro-3-hydroxyanthranilic acid (DHHA) is an important intermediate to synthesize a wide variety of secondary metabolites with potential biological activities. Recently, we engineered a biosynthetic pathway in Pseudomonas chlororaphis GP72 and optimized the batch- fermentation conditions to synthesize DHHA. Herein, we developed a proficient method for the separation and purification of DHHA from the culture extract of recombinant Pseudomonas chlororaphis GP72 by applying macroporous cation-exchange resins. The performance and absorption characteristics of six resins namely D001, HZD-2, JK006, HD-8, HZ016, and 732 were evaluated. Notably, HD-8 resin displayed the pronounced adsorption and desorption capacities for DHHA as compared to others and its equilibrium adsorption data were well-fitted to the Langmuir isotherm. Moreover, HD-8 column chromatography-based dynamic adsorption and desorption tests were carried out to standardize the operational parameters for DHHA separation and purification. Under the optimized separating conditions, the average adsorption capacity and desorption ratio of DHHA was 40.3 ± 0.7 mg/g dry resin and 86.7 ± 2.8%, respectively. The purity and recovery of DHHA was recorded to be 95.3 ± 1.2% and 83.2 ± 2.3%, respectively. Taken together, the HD-8 resin appeared as a promising adsorbent for high-level separation and purification of DHHA from the complex media. In conclusion, this method displays a high potential for separating and purifying DHHA and other bioactive compounds in high yield from the fermentation broth.

反式-2,3-二氢-3-羟基邻氨基苯甲酸（DHHA）是重要的中间体，可以合成具有潜在生物学活性的多种次级代谢产物。最近，我们在绿假单胞菌GP72中设计了一条生物合成途径，并优化了分批发酵条件以合成DHHA。在这里，我们开发了一种从重组绿假单胞菌培养物提取物中分离和纯化DHHA的有效方法。GP72通过应用大孔阳离子交换树脂。评价了六种树脂，即D001，HZD-2，JK006，HD-8，HZ016和732的性能和吸收特性。值得注意的是，HD-8树脂对DHHA的吸附和解吸能力明显优于其他树脂，并且其平衡吸附数据非常适合Langmuir等温线。此外，进行了基于HD-8柱色谱的动态吸附和解吸测试，以标准化DHHA分离和纯化的操作参数。在最佳分离条件下，DHHA的平均吸附量和脱附率分别为干树脂40.3±0.7 mg / g和86.7±2.8％。DHHA的纯度和回收率分别为95.3±1.2％和83.2±2.3％。在一起 HD-8树脂似乎是从复杂介质中高效分离和纯化DHHA的有前途的吸附剂。总之，该方法显示了从发酵液中以高收率分离和纯化DHHA和其他生物活性化合物的巨大潜力。