Evaluation of the food grade expression systems NICE and pSIP for the production of 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum
Evaluation of the food grade expression systems NICE and pSIP for the production of 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum
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DOI：
10.1186/2191-0855-3-7
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发表时间：
2013-01-01
期刊：
影响因子：
3.7
通讯作者：
Michlmayr, Herbert
中科院分区：
文献类型：
--
作者：
Kaswurm, Vanja;Tien-Thanh Nguyen;Michlmayr, Herbert
2，5-二酮-D-葡萄糖酸还原酶(2，5-DKG还原酶)催化2，5-二酮-D-葡萄糖酸(2，5-DKG)还原为2-酮-L古龙酸(2-KLG)，是L抗坏血酸(维生素C)的直接前体(内酯)。该反应是从D-葡萄糖或D-葡萄糖酸生物催化生产食品补充剂维生素C的重要步骤。由于2，5-DKG还原酶通常是重组生产的，因此建立一种既能满足食品安全要求又能生产2，5-DKG还原酶的高效生产工艺是很有意义的。在本研究中，对最近报道的基于乳杆菌的表达系统PSIP(植物乳杆菌)和NICE(乳球菌)的三个食品级变体对谷氨酸棒杆菌生产2，5-DKG还原酶的效率进行了评估。我们的结果表明，两种体系都适合表达2，5-DKG还原酶。1b的产量最高。在pH为6.5的条件下，Plantarum/pSIP609的产酶能力较强。每升发酵液中有262U，这是迄今为止报道的谷氨酸杆菌2，5-DKG还原酶的最高异源表达水平。相应地，lb。Plantarum/pSIP609可能是工业生产2，5-DKG还原酶的大肠杆菌表达系统的一个有趣的替代品。
2,5-diketo-D-gluconic acid reductase (2,5-DKG reductase) catalyses the reduction of 2,5-diketo-D-gluconic acid (2,5-DKG) to 2-keto-L-gulonic acid (2-KLG), a direct precursor (lactone) of L-ascorbic acid (vitamin C). This reaction is an essential step in the biocatalytic production of the food supplement vitamin C from D-glucose or D-gluconic acid. As 2,5-DKG reductase is usually produced recombinantly, it is of interest to establish an efficient process for 2,5-DKG reductase production that also satisfies food safety requirements. In the present study, three recently described food grade variants of the Lactobacillales based expression systems pSIP (Lactobacillus plantarum) and NICE (Lactococcus lactis) were evaluated with regard to their effictiveness to produce 2,5-DKG reductase from Corynebacterium glutamicum. Our results indicate that both systems are suitable for 2,5-DKG reductase expression. Maximum production yields were obtained with Lb. plantarum/pSIP609 by pH control at 6.5. With 262 U per litre of broth, this represents the highest heterologous expression level so far reported for 2,5-DKG reductase from C. glutamicum. Accordingly, Lb. plantarum/pSIP609 might be an interesting alternative to Escherichia coli expression systems for industrial 2,5-DKG reductase production.