四氢嘧啶吸收缺陷突变株高效制备四氢嘧啶

Construction of ectoine absorption defective mutant for efficient ectoine production

摘要:

摘要:【目的】为进一步提高四氢嘧啶(1，4，5，6-四氢-2-甲基-4-嘧啶羧酸;1，4，5，6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid;ectoine) 合成效率，【方法】利用步移PCR 方法克隆了Halomonas salina DSM 5928T四氢嘧啶特异性转运蛋白( ectoine-specific transporter;TeaABC) 编码基因teaABC，Red 重组技术构建了四氢嘧啶吸收缺陷突变株H． salina DSM 5928T(teaABC－)。【结果】H.salina DSM 5928T (teaABC－)10 L发酵罐的四氢嘧啶发酵，四氢嘧啶总浓度9.10(±0.08) g/L，合成效率为9.93(±0.09)g/L.d。【结论】四氢嘧啶吸收缺陷突变株H.salina DSM 5928T(teaABC－)，解除了四氢嘧啶吸收对其合成的负反馈调节，从而显著提 高了四氢嘧啶合成效率。

关键词:关键词: H.salina DSM 5928T，四氢嘧啶，teaABC，Red 重组，吸收缺陷突变株

Abstract:

Abstract: ［Objective］To further enhance ectoine (1，4，5，6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid) synthesis efficiency．［Methods］ We cloned Halomonas salina DSM 5928T teaABC gene of ectoine-specific transporter TeaABC by walking PCR and constructed ectoine absorption defective mutant H． salina DSM 5928T (teaABC－) by Red recombination technology．［Results］ The total concentration of ectoine and productivity in a 10 L fermentor were 9.10(±0.08)g/L and 9.93(±0.09)g/L.d．［Conclusion］ The ectoine absorption defect mutant H． salina DSM 5928T(teaABC－) compromised the negative feedback regulation of ectoine synthesis，which can significantly improve the efficiency of ectoine production.

Key words:Keywords: H． salina DSM 5928T ，ectoine，teaABC，Red recombination，absorption defective mutant