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中国精品科技期刊2020
胡发根, 沈丽, 吉华兰, 罗林, 王立梅, 齐斌, 朱益波. 重组大肠杆菌全细胞合成苯基乳酸的研究[J]. 食品工业科技, 2015, (09): 147-151. DOI: 10.13386/j.issn1002-0306.2015.09.024
引用本文: 胡发根, 沈丽, 吉华兰, 罗林, 王立梅, 齐斌, 朱益波. 重组大肠杆菌全细胞合成苯基乳酸的研究[J]. 食品工业科技, 2015, (09): 147-151. DOI: 10.13386/j.issn1002-0306.2015.09.024
HU Fa-gen, SHEN Li, JI Hua-lan, LUO Lin, WANG Li-mei, QI Bin, ZHU Yi-bo. Production of phenyllactic acid by using whole-cell recombinant Escherichia coli[J]. Science and Technology of Food Industry, 2015, (09): 147-151. DOI: 10.13386/j.issn1002-0306.2015.09.024
Citation: HU Fa-gen, SHEN Li, JI Hua-lan, LUO Lin, WANG Li-mei, QI Bin, ZHU Yi-bo. Production of phenyllactic acid by using whole-cell recombinant Escherichia coli[J]. Science and Technology of Food Industry, 2015, (09): 147-151. DOI: 10.13386/j.issn1002-0306.2015.09.024

重组大肠杆菌全细胞合成苯基乳酸的研究

Production of phenyllactic acid by using whole-cell recombinant Escherichia coli

  • 摘要: 在E.coli BL21(DE3)中过量表达D-乳酸脱氢酶基因(D-ldh),并优化该重组菌全细胞转化苯丙酮酸钠合成苯基乳酸的条件。通过单因素实验和正交实验优化诱导表达条件,并在此基础上对全细胞转化苯丙酮酸钠合成苯基乳酸进行了优化。结果表明,菌体OD600为1.2时添加IPTG至终浓度0.2mmol/L,25℃诱导4h后收集菌体具有最佳转化活性;最优分批转化条件:p H7.0,8.0g/L苯丙酮酸钠,20g/L葡萄糖,1%(v/v)吐温-80,菌体浓度20g/L(干重),37℃,转速200r/min转化0.5h,苯基乳酸产量和转化率分别达到4.91g/L,56%。在上述优化条件上通过流加苯丙酮酸钠和葡萄糖,经6h转化,苯基乳酸最终产量达到17.23g/L,转化率为54%。研究结果表明该重组大肠杆菌成功转化苯丙酮酸合成苯基乳酸,具有较好的应用前景,为系统化代谢工程改造大肠杆菌生物合成苯基乳酸的进一步研究和应用提供了有用的技术参数。 

     

    Abstract: D- lactate dehydrogenase was expressed in recombinant Escherichia coli BL21( DE3),the expression conditions and bioconversion medium were optimized for the production of PLA with whole- cell transformation.Single factor experiments and orthogonal experiments were used to optimize induction conditions and the transformation conditions with the engineered whole- cells transformation. The optimized induction condition for D- ldh expression was IPTG 0.2mmol / L and inducing for 4h at 25℃ and OD6001.2,the optimized batch reaction conditions in phosphate buffer( p H7.0) were: 8g / L PPA,20 g / L glucose,20 g / L( dry cell weight),1% Tween- 80,37℃,200 r / min for 0.5h with PLA production of 4.91 g / L and conversion ratio of 56%.Based on the above optimized conditions,fed- batch fermentation was conducted by intermittent feed PPA and glucose. After 6h transformation,the final PLA concentration reached 17.23 g / L with the conversion ratio of 54%. Results showed that PPA was efficiently transformed into PLA through engineered E.coli BL21( DE3) / p ET28a- D- ldh.This study not only showed a good industrial application prospect,but also laid a foundation for metabolic engineering of E. coli for the production of PLA.

     

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