Bioconversion of Pinoresinol Diglucoside and Pinoresinol from Substrates in the Phenylpropanoid Pathway by Resting Cells of Phomopsis sp.XP-8.
Bioconversion of Pinoresinol Diglucoside and Pinoresinol from Substrates in the Phenylpropanoid Pathway by Resting Cells of Phomopsis sp.XP-8.
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拟茎点霉 XP-8 静息细胞在苯丙素途径中对底物中的松脂醇二葡萄糖苷和松脂醇进行生物转化
DOI：
10.1371/journal.pone.0137066
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Liu Y
中科院分区：
文献类型：
--
作者：
Zhang Y;Shi J;Liu L;Gao Z;Che J;Shao D;Liu Y
半夏醇二葡萄糖苷（PDG）和半夏醇（Pin）通常由植物细胞通过苯丙素途径产生。本研究揭示了一个相关的途径存在于Phomopsis sp. XP-8，一个产生PDG的真菌菌株分离的杜仲树（Eucomiaulmoides Oliv.）的树皮。在拟茎点霉XP-8中加入0.15g/L葡萄糖后，分别以苯丙氨酸、酪氨酸、亮氨酸、肉桂酸和对香豆酸为底物，生成PDG和Pin。在不存在葡萄糖的情况下没有PDG形成，但是在添加除了亮氨酸之外的所有这些底物之后检测到Pin。在存在葡萄糖的所有系统中，PDG和/或Pin的产生以及苯丙氨酸、肉桂酸或对香豆酸的积累以时间和底物浓度依赖性方式与添加的底物直接相关。在分析添加每种底物后产生的产物后，PDG和Pin生物合成的质量流顺序定义为：葡萄糖到苯丙氨酸，苯丙氨酸到肉桂酸，然后到对香豆酸，最后到Pin或PDG。在生物转化过程中，苯丙烷途径中的四个关键酶的活性也被确定，并与其相应产物的积累。通过拟茎点霉属物种生产PDG比目前使用的基于植物的系统表现出更高的效率和成本效益，并且将为用于医疗应用的PDG和/或Pin的大规模生产铺平道路。
Pinoresinol diglucoside (PDG) and pinoresinol (Pin) are normally produced by plant cells via the phenylpropanoid pathway. This study reveals the existence of a related pathway in Phomopsis sp. XP-8, a PDG-producing fungal strain isolated from the bark of the Tu-chung tree (Eucommiaulmoides Oliv.). After addition of 0.15 g/L glucose to Phomopsis sp. XP-8, PDG and Pin formed when phenylalanine, tyrosine, leucine, cinnamic acid, and p-coumaric acid were used as the substrates respectively. No PDG formed in the absence of glucose, but Pin was detected after addition of all these substrates except leucine. In all systems in the presence of glucose, production of PDG and/or Pin and the accumulation of phenylalanine, cinnamic acid, or p-coumaric acid correlated directly with added substrate in a time- and substrate concentration- dependent manner. After analysis of products produced after addition of each substrate, the mass flow sequence for PDG and Pin biosynthesis was defined as: glucose to phenylalanine, phenylalanine to cinnamic acid, then to p-coumaric acid, and finally to Pin or PDG. During the bioconversion, the activities of four key enzymes in the phenylpropanoid pathway were also determined and correlated with accumulation of their corresponding products. PDG production by Phomopsis sp. exhibits greater efficiency and cost effectiveness than the currently-used plant-based system and will pave the way for large scale production of PDG and/or Pin for medical applications.