PDF(Pubmed)
Abstract：
1,4-diaminobutane is widely used in the industrial production of polymers, pharmaceuticals, agrochemicals and surfactants. Owing to economic and environmental concerns, there has been a growing interest in using microbes to produce 1,4-diaminobutane. However, there is lack of research on the influence of cofactors pyridoxal phosphate (PLP) and NADPH on the synthesis of 1,4-diaminobutane. PLP serves as a cofactor of ornithine decarboxylase in the synthesis of 1,4-diaminobutane. Additionally, the synthesis of 1 mol 1,4-diaminobutane requires 2 mol NADPH, thus necessitating consideration of NADPH balance in the efficient synthesis of 1,4-diaminobutane by Escherichia coli. The aim of this study was to enhance the synthesis efficiency of 1,4-diaminobutane through increasing production of PLP and NADPH. By optimizing the expression of the genes associated with synthesis of PLP and NADPH in E. coli, cellular PLP and NADPH levels increased, and the yield of 1,4-diaminobutane also increased accordingly. Ultimately, using glucose as the primary carbon source, the yield of 1,4-diaminobutane in the recombinant strain NAP19 reached 272 mg/L·DCW, by increased 79% compared with its chassis strain.
摘要：
1,4-二氨基丁烷广泛应用于聚合物的工业生产中,制药，农用化学品和表面活性剂。由于经济和环境问题，使用微生物生产1,4-二氨基丁烷的兴趣日益浓厚。然而,关于辅因子磷酸吡哆醛（PLP）和NADPH对1,4-二氨基丁烷合成的影响的研究还很少。PLP在1,4-二氨基丁烷的合成中充当鸟氨酸脱羧酶的辅因子。此外,1摩尔1,4-二氨基丁烷的合成需要2摩尔NADPH，因此，在大肠杆菌有效合成1,4-二氨基丁烷时，必须考虑NADPH平衡。本研究的目的是通过增加PLP和NADPH的产量来提高1,4-二氨基丁烷的合成效率。通过优化PLP和NADPH合成相关基因在大肠杆菌中的表达,细胞PLP和NADPH水平升高，1,4-二氨基丁烷的收率也相应提高。最终,使用葡萄糖作为主要碳源，重组菌株NAP19中1,4-二氨基丁烷的产量达到272mg/L·DCW，与底盘应变相比，增加了79%。
