Direct production of cadaverine from soluble starch using Corynebacterium glutamicum coexpressing α-amylase and lysine decarboxylase
Direct production of cadaverine from soluble starch using Corynebacterium glutamicum coexpressing α-amylase and lysine decarboxylase
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DOI：
10.1007/s00253-008-1751-4
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发表时间：
2009-02-01
影响因子：
5
通讯作者：
Kondo, Akihiko
中科院分区：
文献类型：
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作者：
Tateno, Toshihiro;Okada, Yusuke;Kondo, Akihiko
在这里，我们展示了利用共表达牛链球菌148α-淀粉酶(AMIA)和大肠杆菌K-12赖氨酸脱羧酶(CADA)的谷氨酸棒杆菌菌株一步法生产身体碱。我们构建了大肠杆菌-C。在高结构性表达(HCE)启动子的控制下产生CADA的谷氨酰胺穿梭载体，并将其转化到分泌淀粉酶的谷氨酰胺转化子中。转基因谷氨酸杆菌表达了CADA和AMYA，并保持了它们的活性。以50g/L可溶性淀粉为唯一碳源，在不添加辅酶5‘-磷酸吡哆醛的条件下进行尸素发酵。共表达AmyA和CADA的谷氨酸单胞菌成功地从可溶性淀粉中合成了卡维林，21h后产量为23.4 mM，推测在hce启动子控制下的CADA表达水平足以将L赖氨酸转化为卡维林，因为在发酵过程中培养基中没有积累l-赖氨酸。因此，我们证明了谷氨酸菌利用生物质资源生产身体素的巨大潜力。
Here, we demonstrated the one-step production of cadaverine from starch using a Corynebacterium glutamicum strain coexpressing Streptococcus bovis 148 alpha-amylase (AmyA) and Escherichia coli K-12 lysine decarboxylase (CadA). We constructed the E. coli-C. glutamicum shuttle vector, which produces CadA under the control of the high constitutive expression (HCE) promoter, and transformed this vector into C. glutamicum CSS secreting AmyA. The engineered C. glutamicum expressed both CadA and AmyA, which retained their activity. We performed cadaverine fermentation using 50 g/l soluble starch as the sole carbon source without pyridoxal-5'-phosphate, which is the coenzyme for CadA. C. glutamicum coexpressing AmyA and CadA successfully produced cadaverine from soluble starch and the yield of cadaverine was 23.4 mM after 21 h. CadA expression levels under the control of the HCE promoter were assumed to be sufficient to convert l-lysine to cadaverine, as there was no accumulation ofl-lysine in the culture medium during fermentation. Thus, we demonstrated that C. glutamicum has great potential to produce cadaverine from biomass resources.