Characterization of recombinant phenylalanine aminomutase from Taxus chinensis and synthesis of R-β-arylalanine

Abstract

Abstract: Abstract: In order to synthesize high value β-arylalanine, the gene of phenylalanine aminomutase form Taxus chinensis was cloned and expressed in E.coli. The expression vector Pet-sumo-Tcpam was successfully constructed and transferred into E.coli BL21 to express the recombinant enzyme (TcPAM). The electrophoretically pure TcPAM was prepared using affinity chromatography. The results of MS and CD showed that TcPAM could catalyze isomerization of α-arylalanine to R-β-arylalanine. The activity of TcPAM was 4.11 U/mg at optimum condition of 30℃ and pH9.Metal ions K+, Fe2+, Ca2+ and surfactants CTAB,SDS,Triton X-100,Tween 80 had little effect on the activity, and about 90% of activity remained, while Cu2+and Zn2+ had strong inhibitory effect on TcPAM, Furthermore, the α-arylalanines with different groups at benzene ring were used as substrates. The results suggested that the electron-withdrawing group at benzene ring of substrate promote the shift of α-amino group to β site to improve yield of β-aralyalaines, the yield of 4-MeO-β-phenylalanine reached 45 %.

Key words: Keywords: Taxus chinensis, Phenylalanine ainomutase, β-arylalanine

CLC Number:

Q814

Ping SONG. Characterization of recombinant phenylalanine aminomutase from Taxus chinensis and synthesis of R-β-arylalanine[J]. FOOD SCIENCE, 0, (): 0-0.

References

[1]He L Y, Jagtap P G, Kingston D G I, et al.A Common Pharmacophore for Taxol and the Epothilones Based on the Biological Activity of a Taxane Molecule Lacking a C-13 Side Chain[J].Biochemistry, 2000, 39(14):3972-3978
[2]Wu B, Wiktor Szymański, Heberling M M, et al.Aminomutases: mechanistic diversity,biotechnological applications and future perspectives[J].Trends in Biotechnology, 2011, 29(7):352-362
[3]Prabhakaran P C, Woo N T, Yorgey P S, et al.Biosynthesis of blasticidin S from L-alpha-arginine. Stereochemistry in the arginine-2,3-aminomutase reaction.[J].Journal of the American Chemical Society, 1988, 110(17):5785-5791
[4]Yin X, O' Hare T, Gould S J, et al.Identification and cloning of genes encoding viomycin biosynthesis from Streptomyces vinaceus and evidence for involvement of a rare oxygenase[J].Gene, 2003, 312(30):215-224
[5]Kudo F, Miyanaga A, Eguchi T.Biosynthesis of natural products containing β-amino acids[J].Natural Product Reports, 2014, 31(8):1056-1073
[6]Walker K D, Klettke K, Akiyama T, et al.Cloning,Heterologous Expression,and Characterization of a Phenylalanine Aminomutase Involved in Taxol Biosynthesis[J].Journal of Biological Chemistry, 2004, 279(52):53947-53954
[7]Steele C L, Chen Y, Dougherty B A, et al.Purification,cloning,and functional expression of phenylalanine aminomutase: The first committed step in Taxol side-chain biosynthesis[J].Archives of Biochemistry & Biophysics, 2005, 438(1):0-10
[8]Mutatu W, Klettke K L, Foster C, et al.Unusual Mechanism for an Aminomutase Rearrangement: Retention of Configuration at the Migration Termini[J].Biochemistry, 2007, 46(34):9785-9794
[9]Wohlgemuth R.Asymmetric biocatalysis with microbial enzymes and cells[J].Current Opinion in Microbiology, 2010, 13(3):283-292
[10]Grayson J I, Roos J, Osswald S.Development of a Commercial Process for (S)-β-Phenylalanine[J].Organic Process Research & Development, 2011, 15(5):1201-1206
[11]Forro E, Fueloep F.Recent lipase-catalyzed hydrolytic approaches to pharmacologically important beta-and gamma-amino acids[J].Current Medicinal Chemistry, 2012, 19(36):6178-6187
[12]Ratnayake N D, Theisen C, Walter T, et al.Whole-Cell Biocatalytic Production of Variously Substituted β-Aryl- and β-Heteroaryl-β-Amino Acids[J].Journal of Biotechnology, 2016, 217(10):12-21
[13]Mathew S, Nadarajan S P, Sundaramoorthy U, et al.Biotransformation of β-keto nitriles to chiral (S)-β-amino acids using nitrilase and ω-transaminase[J].Biotechnology Letters, 2016, 39(4):535-543
[14]Mathew S, Bea H, Nadarajan S P, et al.Production of chiral β-amino acids using ω-transaminase from Burkholderia graminis[J].Journal of Biotechnology, 2015, 20(196-197):1-8
[15]Mathew S, Jeong S S, Chung T, et al.Asymmetric synthesis of aromatic β-amino acids using ω-transaminase: Optimizing the lipase concentration to obtainthermodynamically unstable β-keto acids[J].Biotechnology Journal, 2016, 11(1):185-190
[16]朱龙宝, 陶玉贵, 葛飞, 等.苯丙氨酸变位酶的制备、表征及催化合成β-芳香丙氨酸[J].高等学校化学学报, 2017, 38(02):206-211
[17]Parmeggiani F, Weise N J, Ahmed S T, et al.Synthetic and therapeutic applications of ammonia-lyases and aminomutases[J].Chemical Reviews, 2018, 118(1):73-118
[18]Huang S X, Lohman J R, Huang T, et al.A new member of the 4-methylideneimidazole-5-one-containing aminomutase family from the enediyne kedarcidin biosynthetic pathway[J].Proceedings of the National Academy of Sciences, 2013, 110(20):8069-8074
[19]Varga A, Gergely Bánóczi, Nagy B, et al.Influence of the aromatic moiety in α- and β-arylalanines on their biotransformation with phenylalanine 2,3-aminomutase from Pantoea agglomerans[J].Rsc Advances, 2016, 6(61):56412-56420
[20]Ratnayake N D, Wanninayake U, Geiger J H, et al.Stereochemistry and Mechanism of a Microbial Phenylalanine Aminomutase[J].Journal of the American Chemical Society, 2011, 133(22):8531-8533
[21]Chesters C, Wilding M, Goodall M, et al.Thermal Bifunctionality of Bacterial Phenylalanine Aminomutase and Ammonia Lyase Enzymes[J].Angewandte Chemie, 2012, 51(18):4344-4348
[22]Jennewein S, Croteau R.Taxol: biosynthesis,molecular genetics,and biotechnological applications[J].Applied Microbiology and Biotechnology, 2001, 57(1-2):13-19
[23]Chirpich T P, Zappia V, Costilow R N, et al.Lysine 2,3-aminomutasePurification and properties of a pyridoxal phosphate and S-adenosylmethionine-activated enzyme[J].Journal of Biological Chemistry, 1970, 245(7):1778-1789
[24] Takeshi Y, Mina O, Kaori Y, et al.Assignment of the CD Cotton Effect to the Chiral Center in Pseurotins, and the Stereochemical Revision of Pseurotin A2.[J].Marine Drugs, 2016, 14(4):74.
[25]朱龙宝, 杨瑾, 葛飞, 等.成团泛菌苯丙氨酸氨基变位酶的性质表征及用于合成β-苯丙氨酸[J].精细化工, 2019, 36(03):449-454