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Direct cadaverine production from cellobiose using β-glucosidase displaying Escherichia coli

机译:使用展示大肠杆菌的β-葡萄糖苷酶直接从纤维二糖生产尸胺

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摘要

In this study, we demonstrate the one-step production of cadaverine (1,5-diaminopentane) from cellobiose using an Escherichia coli strain displaying β-glucosidase (BGL) on its cell surface. L-lysine decarboxylase (CadA) derived from E. coli and BGL from Thermobifida fusca YX (Tfu0937) fused to the anchor protein Blc from E. coli were co-expressed using E. coli as a host. The expression of CadA was confirmed by Western blotting and BGL activity on the cell surface was evaluated using pNPG as a substrate. Growth on cellobiose as the sole carbon source was also achieved. The OD600 value of the BGL and CadA co-expressing strain was 8.0 after 48 h cultivation, which is higher than that obtained by growth on glucose (5.4 after 48 h cultivation). The engineered strain produced cadaverine from cellobiose more effectively than from glucose: 6.1 mM after 48 h from 28 g/L of consumed cellobiose, vs. 3.3 mM from 20 g/L of consumed glucose.
机译:在这项研究中,我们证明了使用在其细胞表面显示β-葡萄糖苷酶(BGL)的大肠杆菌菌株从纤维二糖一步生产尸胺(1,5-二氨基戊烷)。以大肠杆菌为宿主,共表达源自大肠杆菌的L-赖氨酸脱羧酶(CadA)和来自嗜热栖热菌YX(Tfu0937)的BGL,融合至来自大肠杆菌的锚蛋白Blc。通过蛋白质印迹证实了CadA的表达,并且使用pNPG作为底物评估了细胞表面上的BGL活性。还实现了纤维二糖作为唯一碳源的生长。培养48小时后,BGL和CadA共表达菌株的OD600值为8.0,高于通过葡萄糖生长获得的OD600值(培养48小时后)。该工程菌株从纤维二糖产生的尸胺比从葡萄糖更有效地产生尸胺:48 g / L消耗的纤维二糖在48小时后为6.1 mM,而20 g / L消耗的葡萄糖为3.3 mM。

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