Abstract Ethylenediamine-N,N'-disuccinic acid (EDDS) is a promising chelating agent for the remediation of heavy metal-contaminated soil. In general, EDDS is produced through a chemical method. In this study, we report an efficient biotechnological approach for EDDS production using an immobilized enzyme. We expressed the EDDS lyase in E. coli and obtained 19.8 g/L of EDDS through a reaction catalyzed by crude enzymes, containing EDDS lyase and fumarase. After performing metal affinity chromatography-mediated purification, we thoroughly eliminated the fumarase activity, which could result in the unnecessary formation of malate. Then, the purified EDDS lyase was immobilized on a glutaraldehyde-activated amino carrier, and the immobilized enzyme was used in 11 batches (864.5 h). After optimization, 209.3 g/L EDDS was obtained in a 100 mL reaction system, resulting in 20.2 g of EDDS product with a purity of 99.8 % after isolation. The yields of reaction and isolation were 94.0 % and 91.8 %, respectively. In conclusion, this study describes a promising bioproduction process for industrial-level EDDS production.

中文翻译：

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使用固定化无延胡索酸酶 EDDS 裂解酶生物生产乙二胺-N,N'-二琥珀酸

摘要 乙二胺-N,N'-二琥珀酸（EDDS）是一种很有前景的螯合剂，用于修复重金属污染土壤。一般而言，EDDS 是通过化学方法生产的。在这项研究中，我们报告了一种使用固定化酶生产 EDDS 的有效生物技术方法。我们在大肠杆菌中表达了 EDDS 裂解酶，并通过包含 EDDS 裂解酶和延胡索酸酶的粗酶催化的反应获得了 19.8 g/L 的 EDDS。在进行金属亲和层析介导的纯化后，我们彻底消除了延胡索酸酶活性，这可能导致不必要的苹果酸形成。然后，将纯化的 EDDS 裂解酶固定在戊二醛活化的氨基载体上，固定化酶分 11 批（864.5 h）使用。优化后，209。在 100 mL 反应体系中获得 3 g/L EDDS，得到 20.2 g EDDS 产物，分离后纯度为 99.8%。反应收率和分离收率分别为94.0%和91.8%。总之，本研究描述了一种用于工业级 EDDS 生产的有前途的生物生产工艺。