唾液酸乳糖在婴幼儿配方奶粉产业展现出潜在、广阔的市场前景,但是目前缺少有效、廉价的合成方法。针对该问题,首先构建了2株工程菌JM109(DE3)/pET28a-neuA和JM109(DE3)/pET28a-nst,分别用于合成胞苷单磷酸-N-乙酰神经氨酸(cytidine 5'-monophosphate N-acetylneuraminic acid,CMP-Neu5Ac)和3'-唾液酸乳糖;然后优化了JM109(DE3)/pET28a-neuA合成CMP-Neu5Ac体系;最后通过将啤酒酵母、JM109(DE3)/pET28a-neuA和JM109(DE3)/pET28a-nst耦合发酵,实现了3'-唾液酸乳糖的3菌株全细胞耦合催化合成。由于采用混菌耦合发酵策略来进行3'-唾液酸乳糖的生产,其生产模式不再像酶法一样繁琐和价格高昂,并且实现了CMP-Neu5Ac的高效再生和供给,为经济且规模化生产3'-唾液酸乳糖奠定了技术基础。
游星
,
郭进
,
张洪涛
,
闫竞宇
,
CHAI Wengang
,
周文
,
刘栗彤
,
黎玉
,
吴剑荣
,
詹晓北
. 基于混菌耦合发酵策略合成3'-唾液酸乳糖[J]. 食品与发酵工业, 2021
, 47(17)
: 8
-14
.
DOI: 10.13995/j.cnki.11-1802/ts.027472
Sialyllactose has broad market prospects in infant formula industry. However, it is still a challenge to produce sialyllactose in large-scale. Escherichia coli JM109(DE3)/pET28a-neuA and E. coli JM109(DE3)/pET28a-nst were constructed to synthesize cytidine 5'-monophosphate N-acetylneuraminic acid (CMP-Neu5Ac) and 3'-sialyllactose, respectively. And then the optimized condition of CMP-Neu5Ac synthesis by E. coli JM109(DE3)/pET28a-neuA was developed. Finally, the whole-cell coupling fermentation method with beer yeast, E. coli JM109(DE3)/pET28a-neuA and E. coli JM109(DE3)/pET28a-nst was established to achieve 3'-sialyllactose. The CMP-Neu5Ac can be supplied and efficiently regenerated by the method, which will lay a forceful technical foundation for the economical and large-scale production of 3'-sialyllactose.
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