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首页> 外文期刊>Bioprocess and Biosystems Engineering >Process development for efficient biosynthesis of l-DOPA with recombinant Escherichia coli harboring tyrosine phenol lyase from Fusobacterium nucleatum
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Process development for efficient biosynthesis of l-DOPA with recombinant Escherichia coli harboring tyrosine phenol lyase from Fusobacterium nucleatum

机译：带有从核杆菌中提取酪氨酸酚裂解酶的重组大肠杆菌高效生物合成1-DOPA的工艺开发

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摘要

The tyrosine phenol lyase (TPL) catalyzed synthesis of L-DOPA was regarded as one of the most economic route for L-DOPA synthesis. In our previous study, a novel TPL from Fusobacterium nucleatum (Fn-TPL) was exploited for efficient biosynthesis of L-DOPA. However, the catalytic efficiency decreased when the reaction system expanded from 100 mL to 1 L. As such, the bioprocess for scale-up production of L-DOPA was developed in this study. To increase the stability of substrate and product, as well as decrease the by-product formation, the optimum temperature and pH were determined to be 15 degrees C and pH 8.0, respectively. The initial concentration of pyrocatechol, pyruvate and ammonium acetate was fixed at 8, 5 and 77 g/L and a fed-batch approach was applied with sodium pyruvate, pyrocatechol and ammonium acetate fed in a concentration of 5, 5 and 3.5 g/L, respectively. In addition, L-DOPA crystals were exogenously added to inhibit cell encapsulation by the precipitated product. The final L-DOPA concentration reached higher than 120 g/L with pyrocatechol conversion more than 96% in a 15-L stirred tank, demonstrating the great potential of Fn-TPL for industrial production of L-DOPA.
机译：酪氨酸酚裂解酶（TPL）催化的L-DOPA合成被认为是最经济的L-DOPA合成途径之一。在我们以前的研究中，从核梭状芽胞杆菌（Fn-TPL）获得的新型TPL被用于L-DOPA的有效生物合成。然而，当反应体系从100 mL扩展至1 L时，催化效率下降。因此，本研究开发了用于大规模生产L-DOPA的生物工艺。为了提高底物和产品的稳定性，并减少副产物的形成，最佳温度和pH分别确定为15摄氏度和pH 8.0。丙酮酸邻苯二酚，丙酮酸和乙酸铵的初始浓度固定为8、5和77 g / L，分批补料的方法是分别以5、5和3.5 g / L的浓度添加丙酮酸钠，邻苯二酚和乙酸铵， 分别。另外，外源添加L-DOPA晶体以抑制沉淀产物对细胞的包封。在15升搅拌釜中，最终L-DOPA的浓度达到了120 g / L以上，邻苯二酚的转化率超过96％，这表明Fn-TPL在L-DOPA的工业生产中具有巨大的潜力。

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  • 来源
    《Bioprocess and Biosystems Engineering》 |2018年第9期|1347-1354|共8页
  • 作者

    Tang Xiao-Ling; Liu Xiao; Suo Hui; Wang Zhi-Chao; Zheng Ren-Chao; Zheng Yu-Guo;

  • 作者单位

    Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China;

    Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China;

    Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China;

    Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China;

    Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China;

    Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China;

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  • 正文语种 eng
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  • 关键词

    L-DOPA; Tyrosine phenol lyase; Process development; Fed-batch reaction;

    机译：L-DOPA;酪氨酸酚裂解酶;工艺开发;补料分批反应;

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