Abstract

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.

中文翻译：

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使用固定的l-鼠李糖异构酶和d-庚糖3-表异构酶从d-果糖生产d-阿洛糖

摘要

d -Allose是一种稀有的糖，可以用作多种食品和膳食补充剂的成分，具有营养活性，尤其是出色的抗癌作用，并且可以用于辅助癌症化学疗法和放射疗法等。对于-cost过程d -allose生产，使用的基板一锅酶促过程d -fructose的重组酶，和d -psicose 3-差向异构酶（DPE）和升鼠李糖异构酶（升-RhI）的开发。这些酶是从Ruminococcus sp。克隆的。和枯草芽孢杆菌分别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提取和固定。当反应在5小时后达到平衡时，d-果糖，d-庚糖和d-阿洛糖的质量比为6.6：2.4：1.0。使用低成本的d-果糖底物，可重复使用的固定化酶和一锅法反应，简化了生产过程，降低了生产成本。另外，为了简化酶的提取和固定过程，特别是对于DPE固定化，开发了用于酶捕获和固定的新方法。这是单罐的第一份报告ð使用固定-allose生产升-RhI和DPE。