College of Chemical Engineering, Fujian Engineering Research Center of Advanced Manufacturing Technology for Fine Chemicals, Fuzhou University, Fuzhou 350108, People's Republic of China.
College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
J Agric Food Chem. 2021 Nov 17;69(45):13578-13585. doi: 10.1021/acs.jafc.1c05200. Epub 2021 Nov 4.
d-Allulose is considered an ideal alternative to sucrose and has shown tremendous application potential in many fields. Recently, most efforts on production of d-allulose have focused on enzyme-catalyzed epimerization of cheap hexoses. Here, we proposed an approach to efficiently produce d-allulose through fermentation using metabolically engineered JM109 (DE3), in which a SecY (ΔP) channel and a d-allulose 3-epimerase (DPEase) were co-expressed, ensuring that d-fructose could be transported in its nonphosphorylated form and then converted into d-allulose by cells. Further deletion of , , , , and and the use of Ni in a medium limited the carbon flux flowing into the byproduct-generating pathways and the Embden-Meyerhof-Parnas (EMP) pathway, achieving ≈ 0.95 g/g yield of d-allulose on d-fructose using (DPEase, SecY [ΔP], ΔFruA, ΔManXYZ, ΔMak, ΔGalE, ΔFruK) and 8 μM Ni. In fed-batch fermentation, the titer of d-allulose reached ≈23.3 g/L.
d-阿洛酮糖被认为是蔗糖的理想替代品，在许多领域显示出巨大的应用潜力。最近，生产 d-阿洛酮糖的大部分工作都集中在酶催化廉价己糖的差向异构化上。在这里，我们提出了一种通过使用代谢工程化的 JM109（DE3）发酵来高效生产 d-阿洛酮糖的方法，其中共表达了 SecY（ΔP）通道和 d-阿洛酮糖 3-差向异构酶（DPEase），确保 d-果糖可以以非磷酸化形式运输，然后被细胞转化为 d-阿洛酮糖。进一步缺失和，以及在培养基中使用 Ni 限制了碳通量流入副产物生成途径和 EMP 途径，使用（DPEase、SecY[ΔP]、ΔFruA、ΔManXYZ、ΔMak、ΔGalE、ΔFruK）和 8 μM Ni 时，d-果糖的 d-阿洛酮糖产率约为 0.95 g/g。在分批补料发酵中，d-阿洛酮糖的浓度达到约 23.3 g/L。
