Tyrosol is a pharmacologically active phenolic compound widely used in the medicine and chemical industries.Traditional methods of plant extraction are complicated and chemical synthesis of tyrosol is not commercially...Tyrosol is a pharmacologically active phenolic compound widely used in the medicine and chemical industries.Traditional methods of plant extraction are complicated and chemical synthesis of tyrosol is not commercially viable. In this study, a recombinant Escherichia coli strain was constructed by overexpressing the phenylpyruvate decarboxylase ARO10 from Saccharomyces cerevisiae, which could produce tyrosol from glucose. Furthermore,genes encoding key enzymes from the competing phenylalanine and tyrosine synthesis pathways and the repression protein TyrR were eliminated, and the resulting engineered strain generated 3.57 mmol·L^(-1) tyrosol from glucose. More significantly, codon optimization of ARO10 increased expression and tyrosol titer. Using the novel engineered strain expressing codon-optimized AR10 in shake-flask culture, 8.72 mmol·L^(-1) tyrosol was obtained after 48 h. Optimization of the induction conditions improved tyrosol production to 9.53 mmol·L^(-1)(1316.3 mg·L^(-1)). A higher titer of tyrosol was achieved by reconstruction of tyrosol synthetic pathway in E. coli.展开更多
For the efficient conversion of L-tyrosine(L-Tyr)to tyrosol,which is an aromatic compound widely used in the pharmaceutical and chemical industries,a novel four-enzyme cascade pathway based on the Ehrlich pathway of S...For the efficient conversion of L-tyrosine(L-Tyr)to tyrosol,which is an aromatic compound widely used in the pharmaceutical and chemical industries,a novel four-enzyme cascade pathway based on the Ehrlich pathway of Saccharomyces cerevisiae was designed and reconstructed in Escherichia coli.Then,the expression levels of the relevant enzymes were coordinated using a modular approach and gene duplication after the identification of the pyruvate decarboxylase from Candida tropicalis(CtPDC)as the rate-limiting enzymatic step.In situ product removal(ISPR)strategy with XAD4 resins was explored to avoid product inhibition and further improve tyrosol yield.As a result,the titer and conversion rate of tyrosol obtained were 35.7 g·L^(-1) and 93.6%,respectively,in a 3-L bioreactor.Results presented here provide a potential enzymatic process for industrial production of tyrosol from cheap amino acids.展开更多
建立了一种采用KYA H IQ sil C18柱(250 mm×4.6 mm,5μm),以乙腈-水溶液(体积比1∶9)为流动相,在流速1.0 mL/m in、检测波长280 nm、进样量10μL的条件下,用高效液相色谱同时测定红景天苷及其苷元酪醇含量的方法。利用均匀试验设...建立了一种采用KYA H IQ sil C18柱(250 mm×4.6 mm,5μm),以乙腈-水溶液(体积比1∶9)为流动相,在流速1.0 mL/m in、检测波长280 nm、进样量10μL的条件下,用高效液相色谱同时测定红景天苷及其苷元酪醇含量的方法。利用均匀试验设计研究了分析样品的制备方法,确定了红景天根粉的最佳提取条件为:以蒸馏水为溶剂在60℃下超声波提取50 m in。分别测定了不同产地的红景天属植物中红景天苷和酪醇的含量,西藏产圣地红景天中红景天苷含量最高,达到2.147%,其次为藏产大花红景天和西藏红景天,分别为1.763%和1.271%;四川产大花红景天中酪醇含量最高,为0.298 5%,其次是藏产西藏红景天和大花红景天,分别为0.214 3%和0.183 6%;长鞭红景天和狭叶红景天中2种有效成分的含量均很低,红景天苷含量仅为0.034%和0.003%,酪醇含量则分别为0.016 4%和0.004 8%。展开更多
基金Supported by the Fundamental Research Funds for the Central Universities(JUSRP51611A,JUSRP51504)the Natural Science Foundation of Jiangsu Province(BK20171138)+1 种基金the National High Technology Research and Development Program of China(863 program,2013AA102101-5)the 111 Project(No.1112-06)
文摘Tyrosol is a pharmacologically active phenolic compound widely used in the medicine and chemical industries.Traditional methods of plant extraction are complicated and chemical synthesis of tyrosol is not commercially viable. In this study, a recombinant Escherichia coli strain was constructed by overexpressing the phenylpyruvate decarboxylase ARO10 from Saccharomyces cerevisiae, which could produce tyrosol from glucose. Furthermore,genes encoding key enzymes from the competing phenylalanine and tyrosine synthesis pathways and the repression protein TyrR were eliminated, and the resulting engineered strain generated 3.57 mmol·L^(-1) tyrosol from glucose. More significantly, codon optimization of ARO10 increased expression and tyrosol titer. Using the novel engineered strain expressing codon-optimized AR10 in shake-flask culture, 8.72 mmol·L^(-1) tyrosol was obtained after 48 h. Optimization of the induction conditions improved tyrosol production to 9.53 mmol·L^(-1)(1316.3 mg·L^(-1)). A higher titer of tyrosol was achieved by reconstruction of tyrosol synthetic pathway in E. coli.
基金financially supported by the Fundamental Research Funds for the Central Universities (JUSRP21915)National Natural Science Foundation of China (22008089, 21878126)+2 种基金Provincial Natural Science Foundation of Jiangsu Province(BK20200622)the key technologies Research&Development Program of Jiangsu Province (BE2018623)the National First-Class Discipline Program of Light Industry Technology and Engineering(LITE2018-20)
文摘For the efficient conversion of L-tyrosine(L-Tyr)to tyrosol,which is an aromatic compound widely used in the pharmaceutical and chemical industries,a novel four-enzyme cascade pathway based on the Ehrlich pathway of Saccharomyces cerevisiae was designed and reconstructed in Escherichia coli.Then,the expression levels of the relevant enzymes were coordinated using a modular approach and gene duplication after the identification of the pyruvate decarboxylase from Candida tropicalis(CtPDC)as the rate-limiting enzymatic step.In situ product removal(ISPR)strategy with XAD4 resins was explored to avoid product inhibition and further improve tyrosol yield.As a result,the titer and conversion rate of tyrosol obtained were 35.7 g·L^(-1) and 93.6%,respectively,in a 3-L bioreactor.Results presented here provide a potential enzymatic process for industrial production of tyrosol from cheap amino acids.
文摘建立了一种采用KYA H IQ sil C18柱(250 mm×4.6 mm,5μm),以乙腈-水溶液(体积比1∶9)为流动相,在流速1.0 mL/m in、检测波长280 nm、进样量10μL的条件下,用高效液相色谱同时测定红景天苷及其苷元酪醇含量的方法。利用均匀试验设计研究了分析样品的制备方法,确定了红景天根粉的最佳提取条件为:以蒸馏水为溶剂在60℃下超声波提取50 m in。分别测定了不同产地的红景天属植物中红景天苷和酪醇的含量,西藏产圣地红景天中红景天苷含量最高,达到2.147%,其次为藏产大花红景天和西藏红景天,分别为1.763%和1.271%;四川产大花红景天中酪醇含量最高,为0.298 5%,其次是藏产西藏红景天和大花红景天,分别为0.214 3%和0.183 6%;长鞭红景天和狭叶红景天中2种有效成分的含量均很低,红景天苷含量仅为0.034%和0.003%,酪醇含量则分别为0.016 4%和0.004 8%。
