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Establishing an Artificial Pathway for Efficient Biosynthesis of Hydroxytyrosol
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-01-10 00:00:00 , DOI: 10.1021/acssynbio.7b00385 Xianglai Li 1 , Zhenya Chen 1 , Yifei Wu 1 , Yajun Yan 2 , Xinxiao Sun 1 , Qipeng Yuan 1
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-01-10 00:00:00 , DOI: 10.1021/acssynbio.7b00385 Xianglai Li 1 , Zhenya Chen 1 , Yifei Wu 1 , Yajun Yan 2 , Xinxiao Sun 1 , Qipeng Yuan 1
Affiliation
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Hydroxytyrosol (HT) is a valuable natural phenolic compound with strong antioxidant activity and various physiological and pharmaceutical functions. In this study, we established an artificial pathway for HT biosynthesis. First, efficient enzymes were selected to construct a tyrosol biosynthetic pathway. Aro10 from Saccharomyces cerevisiae was shown to be a better ketoacid decarboxylase than Kivd from Lactococcus lactis for tyrosol production. While knockout of feaB significantly decreased accumulation of the byproduct 4-hydroxyphenylacetic acid, overexpression of alcohol dehydrogenase ADH6 further improved tyrosol production. The titers of tyrosol reached 1469 ± 56 mg/L from tyrosine and 620 ± 23 mg/L from simple carbon sources, respectively. The pathway was further extended for HT production by overexpressing Escherichia coli native hydroxylase HpaBC. To enhance transamination of tyrosine to 4-hydroxyphenylpyruvate, NH4Cl was removed from the culture media. To decrease oxidation of HT, ascorbic acid was added to the cell culture. To reduce the toxicity of HT, 1-dodecanol was selected as the extractant for in situ removal of HT. These efforts led to an additive increase in HT titer to 1243 ± 165 mg/L in the feeding experiment. Assembly of the full pathway resulted in 647 ± 35 mg/L of HT from simple carbon sources. This work provides a promising alternative for sustainable production of HT, which shows scale-up potential.
中文翻译：
建立有效的羟基酪醇生物合成途径
羟基酪醇（HT）是一种有价值的天然酚类化合物，具有很强的抗氧化活性以及各种生理和药学功能。在这项研究中，我们建立了HT生物合成的人工途径。首先，选择有效的酶来构建酪醇生物合成途径。对于酪氨酸生产而言，来自酿酒酵母的Aro10被证明比来自乳酸乳球菌的Kivd更好的酮酸脱羧酶。在淘汰feaB的同时副产物4-羟苯基乙酸的积累显着降低，醇脱氢酶ADH6的过表达进一步提高了酪醇的产量。酪氨酸的酪氨酸滴度达到1469±56 mg / L，简单碳源的滴度达到620±23 mg / L。通过过量表达大肠杆菌天然羟化酶HpaBC，该途径进一步扩展为产生HT 。为了增强酪氨酸向4-羟基苯基丙酮酸的转氨作用，从培养基中除去NH 4 Cl。为了减少HT的氧化，将抗坏血酸添加到细胞培养物中。为了降低HT的毒性，选择了1-十二烷醇作为原位提取剂。去除HT。这些努力导致进料实验中HT滴度的累加值增加至1243±165 mg / L。完整路径的组装产生了647±35 mg / L的HT来自简单的碳源。这项工作为HT的可持续生产提供了一个有希望的替代方法，它显示了扩大规模的潜力。
更新日期：2018-01-10
中文翻译：
建立有效的羟基酪醇生物合成途径
羟基酪醇（HT）是一种有价值的天然酚类化合物，具有很强的抗氧化活性以及各种生理和药学功能。在这项研究中，我们建立了HT生物合成的人工途径。首先，选择有效的酶来构建酪醇生物合成途径。对于酪氨酸生产而言，来自酿酒酵母的Aro10被证明比来自乳酸乳球菌的Kivd更好的酮酸脱羧酶。在淘汰feaB的同时副产物4-羟苯基乙酸的积累显着降低，醇脱氢酶ADH6的过表达进一步提高了酪醇的产量。酪氨酸的酪氨酸滴度达到1469±56 mg / L，简单碳源的滴度达到620±23 mg / L。通过过量表达大肠杆菌天然羟化酶HpaBC，该途径进一步扩展为产生HT 。为了增强酪氨酸向4-羟基苯基丙酮酸的转氨作用，从培养基中除去NH 4 Cl。为了减少HT的氧化，将抗坏血酸添加到细胞培养物中。为了降低HT的毒性，选择了1-十二烷醇作为原位提取剂。去除HT。这些努力导致进料实验中HT滴度的累加值增加至1243±165 mg / L。完整路径的组装产生了647±35 mg / L的HT来自简单的碳源。这项工作为HT的可持续生产提供了一个有希望的替代方法，它显示了扩大规模的潜力。
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