Development of efficient resin-based method for purifying 3-hydroxyphenazine-1-carboxylic acid from Aspergillus sclerotiorum ASJ82 crude extract.

The new metabolite 3-hydroxyphenazine 1-carboxylic acid (3-OH-PCA) was obtained by Aspergillus sclerotiorum ASJ82 whole-cell biocatalysis of phenazine 1-carboxylic acid. Despite the biotechnological perspective, the lack of favorable methods for 3-OH-PCA separation and purification from complex biotransformation media has considerably hampered further experimental studies for large-scale industrial applications because of insufficient amounts of purified compounds. In this context, the present work aimed to develop a simple and proficient technique for the separation and purification of 3-OH-PCA from the crude extraction and culture medium of A. sclerotiorum ASJ82. The static adsorption and desorption characteristics of five macroporous resins were evaluated, namely, HP-20, HZ-818, HZ-801, D001, and D155. Remarkably, compared with the other resins, the HZ-818 resins exhibited notable advantages in the adsorption and desorption of 3-OH-PCA. Additionally, the equilibrium adsorption data of the HZ-818 resins were assessed via the Langmuir and Freundlich isotherm models. The analysis revealed that the experimental data exhibited a favorable fit with the Langmuir isotherm model, providing valuable insights into the adsorption behavior of 3-OH-PCA on the HZ-818 resins. Furthermore, a chromatographic column packed with HZ-818 resins was employed, and dynamic adsorption and desorption experiments were conducted. These tests aimed to establish and optimize the operational parameters required to effectively separate and purify 3-OH-PCA. Under the optimized separation and purification conditions, the average adsorption capacity and desorption ratio of 3-OH-PCA were 21.1 ± 1.2 mg/g dry resin and 88 ± 1.2 %, respectively. After treatment with HZ-818 resin, the 3-OH-PCA content considerably increased. These findings revealed that the purity of 3-OH-PCA was 96.5 ± 2.1 %, whereas the recovery rate was 85.32 ± 1.2 %. The results indicate that the HZ-818 resin has significant potential as an adsorbent for effectively separating and purifying 3-OH-PCA from complex media. Overall, this methodology demonstrates great promise for achieving high yields in the separation and purification of 3-OH-PCA and could be utilized in the large-scale production of 3-OH-PCA purification from A. sclerotiorum ASJ82 extracts in industry, presenting an avenue for further investigation and application.