强化前体(UDP-GalNAc)合成路径提高果糖软骨素的生产

Improving Fructosylated Chondroitin Production by Enhancement of UDP-GalNAc Synthetic Pathway

硫酸软骨素是一种重要的糖胺聚糖,广泛应用于医药、食品、保健品行业。为了提高硫酸软骨素类似物果糖软骨素的生产,通过过量表达磷酸葡糖胺变位酶(GlmM)、葡萄糖胺合酶(GlmS),优化融合蛋白glmM-glmS的表达水平,获得了最优工程菌株E.coli K4-H-glmM-glmS,并确定了最适IPTG浓度(0.4 mmol/L)以及诱导温度(37℃)。最后,在5-L罐水平下,借助DO-stat补料模式,果糖软骨素的产量达到了3.99 g/L,较原始菌株提高了108.9%,为工业化生产果糖软骨素奠定了基础。

Sulfated chondroitin is an important glycosaminoglycans,which is widely used in medical,food and cosmetic fields.To improve the production of fructosylated chondroitin,GlcN synthase(GlmM)and GlcN-6-P synthase(GlmS)were overexpressed,the confusion protein was constructed and a recombinant strain E.coli K4-H-glmM-glmS was obtained.Then,the optimal induction conditions including IPTG concentration and induction temperature were estalished.Finally,with the DO-stat feeding strategy in a 5 L fermenter,the yield was up to 3.99 g/L,which was 108.90%higher than that with wild type strain.The study provides a basis for industrial-scale production of fructosylated chondroitin.