Production of d-allose from d-fructose using immobilized l-rhamnose isomerase and d-psicose 3-epimerase
Production of d-allose from d-fructose using immobilized l-rhamnose isomerase and d-psicose 3-epimerase
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DOI：
10.1007/s00449-019-02262-y
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发表时间：
2019-12-03
影响因子：
3.8
通讯作者：
Lin, Jianqiang
中科院分区：
文献类型：
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作者：
Li, Can;Gao, Ling;Lin, Jianqiang
d-Allose是一种稀有糖，可作为多种食品和膳食补充剂的成分，具有消化活性，尤其具有优异的抗癌作用，用于辅助癌症化疗和放疗等。为了开发一种简单、低成本的生产d-醛糖的工艺，以d-果糖为底物，采用重组酶d-甘露糖3- epimase （DPE）和l-鼠李糖异构酶（l-RhI）一锅法生产d-醛糖。这些酶分别从Ruminococcus sp.和B. subtilis中克隆，在大肠杆菌中成功表达，分别用阴离子交换树脂和氨基树脂进行提取和固定化。反应5h后，d-果糖、d-甘露糖和d-醛糖的质量比为6.6:2.4:1.0。采用低成本的d-果糖底物、可重复使用的固定化酶和一锅反应，简化了生产工艺，降低了生产成本。此外，为了简化酶的提取和固定化过程，开发了新的酶捕获和固定化方法，特别是DPE固定化方法。这是首次报道使用固定化l-RhI和DPE进行一罐d-allose生产。
d-Allose is a rare sugar, can be used as an ingredient in a range of foods and dietary supplements, has alimentary activities, especially excellent anti-cancer effects and used in assisting cancer chemotherapy and radiotherapy, etc. To develop a simple and low-cost process for d-allose production, a one-pot enzymatic process using the substrate of d-fructose, and the recombinant enzymes of d-psicose 3-epimerase (DPE) and l-rhamnose isomerase (l-RhI) was developed. These enzymes were cloned from Ruminococcus sp. and B. subtilis, respectively, successfully expressed in E. coli, extracted and immobilized using anion exchange resin and amino resin, respectively. The mass ratio of d-fructose, d-psicose and d-allose was 6.6:2.4:1.0 when the reaction reached equilibrium after 5 h of reaction. Using the low-cost substrate of d-fructose, the reusable immobilized enzymes and the one-pot reaction, the production process is simplified and the production cost is decreased. In addition, to simplify the enzyme extraction and immobilization processes, new methods for enzyme capture and immobilization were developed especially for DPE immobilization. This is the first report for one-pot d-allose production using immobilized l-RhI and DPE.