陈永涛, 柯崇榕, 杨欣伟, 黄建忠. 重组大肠杆菌全细胞催化合成四氢嘧啶[J]. 福建农业科技, 2020, (10): 10-15. DOI: 10.13651/j.cnki.fjnykj.2020.10.002
    引用本文: 陈永涛, 柯崇榕, 杨欣伟, 黄建忠. 重组大肠杆菌全细胞催化合成四氢嘧啶[J]. 福建农业科技, 2020, (10): 10-15. DOI: 10.13651/j.cnki.fjnykj.2020.10.002
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    CHEN Yong-tao, KE Chong-rong, YANG Xin-wei, HUANG Jian-zhong. Synthesis of Ectoine by Whole-cell Catalysis from Recombinant Escherichia Coli[J]. Fujian Agricultural Science and Technology, 2020, (10): 10-15. DOI: 10.13651/j.cnki.fjnykj.2020.10.002
    Citation: CHEN Yong-tao, KE Chong-rong, YANG Xin-wei, HUANG Jian-zhong. Synthesis of Ectoine by Whole-cell Catalysis from Recombinant Escherichia Coli[J]. Fujian Agricultural Science and Technology, 2020, (10): 10-15. DOI: 10.13651/j.cnki.fjnykj.2020.10.002
    shu

    重组大肠杆菌全细胞催化合成四氢嘧啶

    Synthesis of Ectoine by Whole-cell Catalysis from Recombinant Escherichia Coli

      摘要
    • 摘要: 四氢嘧啶是天冬氨酸的环状氨基酸衍生物,广泛应用于化妆品、食品、药品等领域。通过比较了5个不同来源的四氢嘧啶合成基因簇ectABC,发现从专性嗜碱芽孢杆菌(Bacillus pseudofirmusOF4)来源的基因簇整合到大肠杆菌BW25113中后,得到的重组菌pYB1s-BPectABC/BW25113产四氢嘧啶能力最好。利用单因素试验初步优化了pYB1s-BPectABC/BW25113全细胞催化条件:以100 mmol·L-1天冬氨酸钠和100 mmol·L-1甘油为底物,在100 mmol·L-1 KCl的转化液(pH=7.0)中,30℃下反应24 h,最终四氢嘧啶的产量可达3.10 g·L-1,转化产率达到0.129 g·L-1·h-1。本研究初步得到了四氢嘧啶合成重组菌pYB1s-BPectABC/BW25113,为后续四氢嘧啶的研究奠定了基础。

       

      Abstract: Ectoine is a cyclic amino acid derivative of aspartic acid, which is widely used in the fields such as cosmetics, food, medicine and so on. In this paper, by comparing the synthetic gene clusters ectABC of ectoine from five different sources, it was found that the recombinant strain pYB1s-BPectABC/BW25113 obtained from Bacillus pseudofirmus OF4 had the best ability to produce ectoine when it was integrated into Escherichia coli BW25113. The conditions for whole-cell catalysis of pYB1s-BPectABC/BW25113 were preliminarily optimized by single factor experiment:the pYB1s-BPectABC/BW25113 was reacted with 100 mmol·L-1 sodium aspartate and 100 mmol·L-1 glycerine as the substrates in the conversion solution (pH=7.0) of 100 mmol·L-1 KCl for 24h at 30℃, and the final production of ectoine reached 3.10 g·L-1, and the transformation efficiency reached 0.129 g·L-1·h-1. In this experiment, the ectoine synthetic recombinant strain pYB1s-BPectABC/BW25113 was preliminarily obtained, which laid a foundation for the subsequent researches on ectoine.

       

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