Cloning And Functional Identification Of Tyrosine Decarboxylase Gene From Cistanche Tubulata
Apr 07, 2023
Cistanche tubulosa (Schrenk) Wish is a perennial herbaceous parasitic plant of the genus Cistanche in the family Lewandaceae. It is a precious traditional Chinese medicine with a sweet, salty, and warm nature. It has functions such as tonifying kidney yang, benefiting essence and blood, moistening intestines, and defecating. Phenylethanol glycosides are its main active ingredients.
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These compounds have diverse structures and extensive pharmacological activities, making them an important source of lead compounds for medicinal activity. However, the natural sources of these compounds are limited, the chemical synthesis route is cumbersome, the reaction conditions are harsh, and the cost is high, which cannot be met for commercial production.
Main Chemical Constituents of Cistanche deserticola
And explore the key enzyme genes involved in the biosynthesis pathway of phenylethanoid glycosides, and through genetic engineering,The method can provide the possibility for enzymatic synthesis of structurally diverse phenylethanoid glycosides.
Phenylethanol glycoside is the main active component of Cistanche deserticola
The research group conducted preliminary research on glycosyltransferases and acyltransferases modified after catalyzing the biosynthesis of tyrosols (key intermediates of phenylethanoid glycosides). The synthesis of tyrosol requires the use of tyrosine as a precursor, catalyzed by tyrosine decarboxylase to produce tyramine, which is then generated through a series of reactions such as monoamine oxidase and alcohol dehydrogenase. Catalyzed by tyrosine decarboxylase the decarboxylation of tyrosine is the key step of its initial reaction.
Cistanche deserticola experiment
In this study, a tyrosine decarboxylase gene TDC was cloned from the genome of Cistanche tubulosa by RT-PCR method combined with RACE technology. The gene-encoding protein contains 509 amino acids, which was successfully expressed in E. coli, and the recombinant protein with a molecular weight of 61 kDa was isolated and purified.
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The in vitro functional identification results showed that TDC can catalyze the decarboxylation of tyrosine (1) to produce tyramine (2), and the product was identified by HR-MS and reference substance control. Further research was conducted on the subcellular localization, optimal catalytic conditions, and enzymatic selectivity of the recombinant protein. This gene can be used as an important biosynthetic element in the construction of engineering bacteria for the biosynthesis of phenylethanolamine glycosides, and for the enzymatic synthesis of structurally diverse phenylethanolamine glycosides Lay the foundation.
reference:
(1) Song Zhihong, Lei Li, Tu Pengfei, Chinese herbal medicine, 2003,9113-115
(2) Song Zhihong, Tu Pengfei, Zhao Yuying, Zheng Junhua, Chinese herbal medicine, 2000,11,10-12
