Structure and Evolution of Alanine/serine Decarboxylases and the Engineering of Theanine Production

Overview

Journal Elife

Specialty Biology

Date 2024 Sep 17

PMID 39287621

Authors

Hao Wang

Biying Zhu

Siming Qiao

Chunxia Dong

Xiaochun Wan

Weimin Gong

Zhaoliang Zhang

Affiliations

Soon will be listed here.

Abstract

Ethylamine (EA), the precursor of theanine biosynthesis, is synthesized from alanine decarboxylation by alanine decarboxylase (AlaDC) in tea plants. AlaDC evolves from serine decarboxylase (SerDC) through neofunctionalization and has lower catalytic activity. However, lacking structure information hinders the understanding of the evolution of substrate specificity and catalytic activity. In this study, we solved the X-ray crystal structures of AlaDC from (CsAlaDC) and SerDC from (AtSerDC). Tyr of AtSerDC or the corresponding Tyr of CsAlaDC is essential for their enzymatic activity. Tyr of AtSerDC and the corresponding Phe of CsAlaDC determine their substrate specificity. Both CsAlaDC and AtSerDC have a distinctive zinc finger and have not been identified in any other Group II PLP-dependent amino acid decarboxylases. Based on the structural comparisons, we conducted a mutation screen of CsAlaDC. The results indicated that the mutation of L110F or P114A in the CsAlaDC dimerization interface significantly improved the catalytic activity by 110% and 59%, respectively. Combining a double mutant of CsAlaDC with theanine synthetase increased theanine production 672% in an system. This study provides the structural basis for the substrate selectivity and catalytic activity of CsAlaDC and AtSerDC and provides a route to more efficient biosynthesis of theanine.

References

Ng J, Papandreou A, Heales S, Kurian M . Monoamine neurotransmitter disorders--clinical advances and future perspectives. Nat Rev Neurol. 2015; 11(10):567-84. DOI: 10.1038/nrneurol.2015.172. View

Haskell C, Kennedy D, Milne A, Wesnes K, Scholey A . The effects of L-theanine, caffeine and their combination on cognition and mood. Biol Psychol. 2007; 77(2):113-22. DOI: 10.1016/j.biopsycho.2007.09.008. View

Torrens-Spence M, von Guggenberg R, Lazear M, Ding H, Li J . Diverse functional evolution of serine decarboxylases: identification of two novel acetaldehyde synthases that uses hydrophobic amino acids as substrates. BMC Plant Biol. 2014; 14:247. PMC: 4177580. DOI: 10.1186/s12870-014-0247-x. View

Chellam Gayathri S, Manoj N . Structural insights into the mechanism of internal aldimine formation and catalytic loop dynamics in an archaeal Group II decarboxylase. J Struct Biol. 2019; 208(2):137-151. DOI: 10.1016/j.jsb.2019.08.009. View

Takagi Y, Kurihara S, Higashi N, Morikawa S, Kase T, Maeda A . Combined administration of (L)-cystine and (L)-theanine enhances immune functions and protects against influenza virus infection in aged mice. J Vet Med Sci. 2009; 72(2):157-65. DOI: 10.1292/jvms.09-0067. View

Torrens-Spence M, Chiang Y, Smith T, Vicent M, Wang Y, Weng J . Structural basis for divergent and convergent evolution of catalytic machineries in plant aromatic amino acid decarboxylase proteins. Proc Natl Acad Sci U S A. 2020; 117(20):10806-10817. PMC: 7245119. DOI: 10.1073/pnas.1920097117. View

Qu L, Gu S, Zhang J, Zhao C, Deng X . [Determination of 18 amino acids in three different kinds of milk powder by ultra performance liquid chromatography coupled with pre-column derivatization]. Se Pu. 2021; 39(5):472-477. PMC: 9404011. DOI: 10.3724/SP.J.1123.2020.07008. View

Yu G, Lam T, Zhu H, Guan Y . Two Methods for Mapping and Visualizing Associated Data on Phylogeny Using Ggtree. Mol Biol Evol. 2018; 35(12):3041-3043. PMC: 6278858. DOI: 10.1093/molbev/msy194. View

Bai P, Wang L, Wei K, Ruan L, Wu L, He M . Biochemical characterization of specific Alanine Decarboxylase (AlaDC) and its ancestral enzyme Serine Decarboxylase (SDC) in tea plants (Camellia sinensis). BMC Biotechnol. 2021; 21(1):17. PMC: 7923638. DOI: 10.1186/s12896-021-00674-x. View

10.

Khan M, Nawaz W . The emerging roles of human trace amines and human trace amine-associated receptors (hTAARs) in central nervous system. Biomed Pharmacother. 2016; 83:439-449. DOI: 10.1016/j.biopha.2016.07.002. View

11.

Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J, Bealer K . BLAST+: architecture and applications. BMC Bioinformatics. 2009; 10:421. PMC: 2803857. DOI: 10.1186/1471-2105-10-421. View

12.

Yunus I, Liu Y, Nakamura Y . The importance of SERINE DECARBOXYLASE1 (SDC1) and ethanolamine biosynthesis during embryogenesis of Arabidopsis thaliana. Plant J. 2016; 88(4):559-569. DOI: 10.1111/tpj.13278. View

13.

LARKIN M, Blackshields G, Brown N, Chenna R, McGettigan P, McWilliam H . Clustal W and Clustal X version 2.0. Bioinformatics. 2007; 23(21):2947-8. DOI: 10.1093/bioinformatics/btm404. View

14.

Lin J, Wang Y, Chen L, Yang Y, Tu Z, Ye Y . Effect of the Presence of Stem on Quality of Oolong Tea. Foods. 2022; 11(21). PMC: 9657438. DOI: 10.3390/foods11213439. View

15.

Komori H, Nitta Y, Ueno H, Higuchi Y . Structural study reveals that Ser-354 determines substrate specificity on human histidine decarboxylase. J Biol Chem. 2012; 287(34):29175-83. PMC: 3436558. DOI: 10.1074/jbc.M112.381897. View

16.

Liu Q, Duan H, Luan J, Yagasaki K, Zhang G . Effects of theanine on growth of human lung cancer and leukemia cells as well as migration and invasion of human lung cancer cells. Cytotechnology. 2009; 59(3):211-7. PMC: 2774573. DOI: 10.1007/s10616-009-9223-y. View

17.

Dong C, Li F, Yang T, Feng L, Zhang S, Li F . Theanine transporters identified in tea plants (Camellia sinensis L.). Plant J. 2019; 101(1):57-70. DOI: 10.1111/tpj.14517. View

18.

Lin S, Chen Z, Chen T, Deng W, Wan X, Zhang Z . Theanine metabolism and transport in tea plants ( L.): advances and perspectives. Crit Rev Biotechnol. 2022; 43(3):327-341. DOI: 10.1080/07388551.2022.2036692. View

19.

Schmittgen T, Zakrajsek B, Mills A, Gorn V, Singer M, Reed M . Quantitative reverse transcription-polymerase chain reaction to study mRNA decay: comparison of endpoint and real-time methods. Anal Biochem. 2000; 285(2):194-204. DOI: 10.1006/abio.2000.4753. View

20.

Madeira F, Pearce M, Tivey A, Basutkar P, Lee J, Edbali O . Search and sequence analysis tools services from EMBL-EBI in 2022. Nucleic Acids Res. 2022; 50(W1):W276-W279. PMC: 9252731. DOI: 10.1093/nar/gkac240. View