Structural Insights Into The Proteions Of FMT And AtNSP

Due to the prevalence of lignin, plants are difficult to be break down and exploit in industrial production, especially in the production of bioenergy. In order to make better use of plants, reduce the energy consumption of industrial production, the transformation of lignin makes it easier to be chemically disassembled into a popular research direction. The latest research found that lignin monomer-ferulic acid salt transferase (FMT) can be introduced into the lignin polymer in the poplar pigment, making the lignin easier to be decomposed and utilized, reducing energy consumption and having good Application prospects. Experiments show that DNA was extracted from Angelica sinensis and the FMT-related gene was cloned and transferred to the genome of the plant. The final synthetic lignin contains ferulic acid.We have successfully built the AsFMT (1-441 aa) cloning, and make it expressed in the E. coli successfully by connecting GST tag with protein. By the expression of protein purification, we have got the AsFMT protein. Through crystallization experiments, we finally won eight crystal growth conditions,after a series of optimization, we obtained a high quality of protein crystal.Another part of this paper is about the structural insight into Arabidopsis thaliana NSP protein. There is an important metabolite of glucosinolates in the cruciferous plants. Myrosinase can hydrolyze glucosinolates. The metabolic process of glucosinolate is related to some specifier proteins. The metabolites of different specifier proteins are different. Up to now, identified specifier proteins include epithiospecifier proteins (ESPs), thiocyanate forming proteins(TFPs), and nitrile-specifier proteins (NSPs),only the structure and catalytic mechanism of NSPs have not yet been resolved, the structure and function of NSP protein were researched in this paper. The recombinant plasmid pET-22b(+)was constructed and the target protein was expressed by prokaryotic E. coli expression system. After the high quality crystal was obtained, we combined X-ray diffraction data with molecular replacement,finally we have got three-dimensional structure of the protein for 3.02 A in diffraction.Molecular docking studies revealed the different conformation of R157 in AtNSPl.The study suggested the different product spectrum mechanism of specifier proteins.