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Phomopsis sp. XP-8
Scientific Reports · 2019 ·Vol. 9 ·期 1 ·1-14 · IF 4.011

Tracing the mass flow from glucose and phenylalanine to pinoresinol and its glycosides in Phomopsis sp. XP-8 using stable isotope assisted TOF-MS

Zhang Yan,Shi Junling,Ni Yongqing,Liu Yanlin,Zhao Zhixia,Zhao Xixi,Gao Zhenhong

Phomopsis sp. XP-8, an endophytic fungus from the bark of Tu-Chung ( Eucommia ulmoides Oliv) showed capability to biosynthesize pinoresinol (Pin) and pinoresinol diglucoside (PDG) from glucose (glu) and phenylalanine (Phe). To verify the mass flow in the biosynthesis pathway, [ 13 C 6 ]-labeled glu and [ 13 C 6 ]-labeled Phe were separately fed to the strain as sole substrates and [ 13 C 6 ]-labeled products were detected by ultra-high-performance liquid chromatography-quadrupole time of flight mass spectrometry. As results, [ 13 C 6 ]-labeled Phe was incorporated into [ 13 C 6 ]-cinnamylic acid (Ca) and p -coumaric acid ( p -Co), and [ 13 C 12 ]-labeled Pin, which revealed that the Pin benzene ring came from Phe via the phenylpropane pathway. [ 13 C 6 ]-Labeled Ca and p -Co, [ 13 C 12 ]-labeled Pin, [ 13 C 18 ]-labeled pinoresinol monoglucoside (PMG), and [ 13 C 18 ]-labeled PDG products were found when [ 13 C 6 ]-labeled glu was used, demonstrating that the benzene ring and glucoside of PDG originated from glu. It was also determined that PMG was not the direct precursor of PDG in the biosynthetic pathway. The study identified the occurrence of phenylalanine- lignan biosynthesis pathway in fungi at the level of mass flow.

研究领域: 生物化学 · 代谢组学 · 植物次生代谢
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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY · 2015 ·Vol. 99 ·期 11 ·4629-4643 · IF 3.337

Production of pinoresinol diglucoside, pinoresinol monoglucoside, and pinoresinol by Phomopsis sp. XP-8 using mung bean and its major components

Zhang Yan,Shi Junling,Gao Zhenhong,Yangwu Ruiming,Jiang Huanshi,Che Jinxin,Liu Yanlin

Phomopsis sp. XP-8 is an endophytic fungus that has the ability to produce pinoresinol diglucoside (PDG) in vitro and thus has potential application for the biosynthesis of PDG independent of plants. When cultivated in mung bean medium, PDG production was significantly improved and pinoresinol monoglucoside (PMG) and pinoresinol (Pin) were also found in the culture medium. In this experiment, starch, protein, and polysaccharides were isolated from mung beans and separately used as the sole substrate in order to explore the mechanism of fermentation and identify the major substrates that attributed to the biotransformation of PDG, PMG, and Pin. The production of PDG, PMG, and Pin was monitored using high-performance liquid chromatography (HPLC) and confirmed using HPLC-MS. Activities of related enzymes, including phenylalanine ammonia-lyase (PAL), trans-cinnamate 4-hydroxylase (C4H), and 4-coumarate-CoA ligase (4CL) were analyzed and tracked during the cultivation. The reaction system contained the compounds isolated from mung bean in the designed amount. Accumulation of phenylalanine, cinnamic acid, p-coumaric acid, PDG, PMG, and Pin and the activities of PAL, C4H, and 4CL were measured during the bioconversion. PMG was found only when mung bean polysaccharide was analyzed, while production of PDG and Pin were found when both polysaccharide and starch were analyzed. After examining the monosaccharide composition of the mung bean polysaccharide and the effect of the different monosaccharides had on the production of PMG, PDG, and Pin, galactose in mung bean polysaccharide proved to be the major factor that stimulates the production of PMG.

研究领域: 微生物学 · 生物化学 · 天然产物化学
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PLoS One · 2015 ·Vol. 10 ·期 9 ·e0137066 · IF 3.234

Bioconversion of Pinoresinol Diglucoside and Pinoresinol from Substrates in the Phenylpropanoid Pathway by Resting Cells of Phomopsis sp.XP-8

Yan Zhang,Junling Shi,Laping Liu,Zhenhong Gao,Jinxin Che,Dongyan Shao,Yanlin Liu

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.

研究领域: 微生物学 · 生物化学 · 药理学
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