Department of Life Science and Biochemical Engineering, Institute of Biomolecule Reconstruction (iBR), Sun Moon University, 70 Sunmoon-ro 221, Tangjeong-myeon, Asan-si, Chungnam, 31460, Republic of Korea.
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, 70 Sunmoon-ro 221, Tangjeong-myeon, Asan-si, Chungnam, 31460, Republic of Korea.
World J Microbiol Biotechnol. 2018 May 23;34(6):77. doi: 10.1007/s11274-018-2462-3.
Azasugars, such as 1-deoxynojirymicin (1-DNJ), are associated with diverse pharmaceutical applications, such as antidiabetic, anti-obesity, anti-HIV, and antitumor properties. Different azasugars have been isolated from diverse microbial and plant sources though complicated purification steps, or generated by costly chemical synthesis processes. But the biosynthesis of such potent molecules using Escherichia coli as a heterologous host provides a broader opportunity to access these molecules, particularly by utilizing synthetic biological, metabolic engineering, and process optimization approaches. This work used an integrated approach of synthetic biology, enzyme engineering, and pathway optimization for rational metabolic engineering, leading to the improved production of 1-DNJ. The production of 1-DNJ in recombinant E. coli culture broth was confirmed by enzymatic assays and mass spectrometric analysis. Specifically, the pathway engineering for its key precursor, fructose-6-phosphate, along with optimized media condition, results in the highest production levels. When combined, 1-DNJ production was extended to ~ 273 mg/L, which is the highest titer of production of 1-DNJ reported using E. coli.
氮杂糖，如 1-脱氧野尻霉素（1-DNJ），具有广泛的药用价值，如抗糖尿病、抗肥胖、抗 HIV 和抗肿瘤特性。不同的氮杂糖已从不同的微生物和植物来源中分离出来，尽管需要经过复杂的纯化步骤，或者通过昂贵的化学合成过程来产生。但是，使用大肠杆菌作为异源宿主来生物合成这些有效分子提供了更多获得这些分子的机会，特别是通过利用合成生物学、代谢工程和过程优化方法。这项工作采用了合成生物学、酶工程和途径优化的综合方法，进行合理的代谢工程，从而提高了 1-DNJ 的产量。通过酶促测定和质谱分析确认了 1-DNJ 在重组大肠杆菌培养物中的产生。具体来说，通过对其关键前体果糖-6-磷酸的途径工程以及优化的培养基条件，可实现最高的产量水平。两者结合使用，1-DNJ 的产量可扩展至~273mg/L，这是使用大肠杆菌报告的 1-DNJ 最高产量。
