文章摘要
三酶偶联全细胞生物合成L-酪氨酸
Biosynthesis of L-Tyrosine with Aldolase, D-Serine Dehydratase and Tyrosine Phenol-Lyase
投稿时间:2018-03-07  修订日期:2018-10-30
DOI:
中文关键词: 酶偶联  醛缩酶  D-丝氨酸脱水酶  酪氨酸酚裂解酶  L-酪氨酸
英文关键词: Three enzyme coupling  Aldolase  D-serine dehydratase  Tyrosine Phenol-Lyase  L-Tyrosine
基金项目:
作者单位E-mail
陈明亮 长兴制药股份有限公司 976608463@qq.com 
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中文摘要:
      通过将醛缩酶(Aldolase,ALD)、D-丝氨酸脱水酶(D-Serine Dehydratase,SDH)和酪氨酸酚裂解酶(Tyrosine Phenol-Lyase,TPL)三酶偶联,催化甘氨酸、甲醛和苯酚合成L-酪氨酸(L-Tyrosine,L-Tyr)。经考察确定了三酶偶联最佳反应条件:pH 8.5,温度35 ℃,6 g/L乙酸铵、0.1 g/L PLP,三种酶细胞质量配比(ALD∶SDH∶TPL)为6∶3∶5。当甘氨酸加量为50 g/L,利用上述工艺进行10000 L转化验证,反应11 h,L-Tyr浓度可达117 g/L,转化率为97%,转化体系放大后转化率提高4 %,收率为85%。三酶偶联生物法合成L-Tyr直接将底物甘氨酸、甲醛和苯酚经酶催化反应获得L-Tyr,节省了D-丝氨酸和丙酮酸铵的提取步骤,对资源的合理利用以及对绿色合成工艺的开发具有重要的参考意义。
英文摘要:
      We used multienzyme coupled catalysis to produce L-Tyrosine(L-Tyr). In this study, Glycine was catalyzed to synthesize D-Serine by Aldolase(ALD), and D-Serine was transferred to pyruvic acid ammonium by D-Serine Dehydratase(SDH). Then, L-Tyr was synthesized by Tyrosine Phenol-Lyase(TPL) when the substrate phenol was added into the reaction system.The optimal reaction conditions were shown as follows: pH=8.5, 35 ℃, 6 g/L of ammonium acetate, 0.1 g/L of PLP, m(ALD):m(SDH):m(TPL)=6∶3∶5. We used the above conditions to catalyze glycine at the concentration of 50 g/L for 11 h, the concentration of L-Tyr was 117 g/L with a conversion of 97%, increasing by 4% compared with small-scale test and the yield reached 85%.The method of biosynthesis of L-Tyr can not only eliminate the purification process of D-Serine and pyruvic acid ammonium, but also has a reference significance for the reasonable utilization of resources and green synthesis process.
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