利用D-阿洛酮糖3-差向异构酶(D-psicose 3-epimerase,DPE)对D-果糖的异构化反应是目前制备D-阿洛酮糖最经济环保的方式。但因DPE贮存稳定性较差,限制了其工业化应用。该文以Clostridium cellulolyticum DPE (CcDPE)为研究对象,寻找绿色高效、成本低廉且不影响后续酶反应的贮存稳定剂。考察了糖类、金属离子与多元醇类等稳定剂对CcDPE贮存效果和对后续酶反应的影响。先进行单因素实验,后对获得的2个最适的稳定剂进行复配。结果表明,甘油和Co2+能显著提高CcDPE粗酶液的贮存稳定性,且不会对后续酶转化反应造成负面影响。在25 ℃下,1 mmol/L Co2+可以将粗酶液半衰期从4.5 d延长到46 d; 30%(体积分数)甘油可以将半衰期延长到33 d; 20%(体积分数)甘油与0.5 mmol/L Co2+的复配组合可以将半衰期延长至80 d。研究结果为CcDPE的进一步工业化应用提供了技术支撑。
The isomerization of D-fructose by D-psicose 3-epimerase (DPE) is the most economical and environmentally friendly way to prepare D-psicose. However, the poor storage stability of DPE limits its industrial application. Clostridium cellulolyticum DPE (CcDPE) was used as the research object to find a green, efficient, low-cost storage stabilizer that does not affect the subsequent enzymatic reaction. The effects of stabilizers such as carbohydrate, metal ions and polyols on the storage effect of CcDPE and the subsequent enzymatic reaction were investigated. First, single factor experiment was carried out, and then the two most suitable stabilizers were compounded. The results showed that glycerol and Co2+ could significantly improve the storage stability of CcDPE crude enzyme solution, and did not have a negative impact on the subsequent enzymatic conversion reaction. At 25 ℃, 1 mmol/L Co2+ could prolong the half-life of crude enzyme from 4.5 d to 46 d. 30% glycerol could prolong the half-life to 33 d; the combination of 20% glycerol and 0.5 mmol/L Co2+ could prolong the half-life to 80 d. The results provide technical support for further industrial application of CcDPE.
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