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Threonine aldolases—screening, properties and applications in the synthesis of non-proteinogenic β-hydroxy-α-amino acids

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Abstract

Threonine aldolases (TAs) constitute a powerful tool for catalyzing carbon–carbon bond formations in synthetic organic chemistry, thus enabling an enantio- and diastereoselective synthesis of β-hydroxy-α-amino acids. Starting from the achiral precursors glycine and an aldehyde, two new stereogenic centres are formed in this catalytic step. The resulting chiral β-hydroxy-α-amino acid products are important precursors for pharmaceuticals such as thiamphenicol, a l-threo-phenylserine derivative or l-threo-3,4-dihydroxyphenylserine. TAs are pyridoxal-5-phosphate-dependent enzymes, which, in nature, catalyze the cleavage of l-threonine or l-allo-threonine to glycine and acetaldehyde in a glycine biosynthetic pathway. TAs from a broad number of species of bacteria and fungi have been isolated and characterised as biocatalysts for the synthesis of β-hydroxy-α-amino acids. In this review, screening methods to obtain novel TAs, their biological function, biochemical characterisation and preparative biotransformations with TAs are described.

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References

  • Alonso M, Riera A (2005) Improved preparation of β-hydroxy-alpha-amino acids: direct formation of sulfates by sulfuryl chloride. Tetrahedron Asymmetr 16:3908–3912

    Article  CAS  Google Scholar 

  • Aoyama Y, Motokawa Y (1981) L-Threonine dehydrogenase of chicken liver. Purification, characterization, and physiological significance. J Biol Chem 256:12367–12373

    CAS  Google Scholar 

  • Baik SH, Yoshioka H (2009) Enhanced synthesis of l-threo-3,4-dihydroxyphenylserine by high-density whole-cell biocatalyst of recombinant l-threonine aldolase from Streptomyces avelmitilis. Biotechnol Lett 31:443–448

    Article  CAS  Google Scholar 

  • Baik SH, Yoshioka H, Yukawa H, Harayama S (2007) Synthesis of L-threo-3,4-dihydroxyphenylserine (L-threo-DOPS) with thermostabilized low-specific L-threonine aldolase from Streptomyces coelicolor A3. J Microbiol Biotechnol 17:721–727

    CAS  Google Scholar 

  • Berkessel A, Gröger H (2005) Asymmetric organocatalysis. Wiley-VCH, Weinheim, p 159

    Book  Google Scholar 

  • Blank S, Seebach D (1993) Preparation of (R,R)-2-amino-3-hydroxycarboxylic or (S,S)-2-amino-3-hydroxycarboxylic acids (allo-threonine analogs) by acylation reduction of t-butyl 2-t-butyl-3-methyl-4-oxoimidazolidine-1-carboxylate (Boc-Bmi). Liebigs Annalen Der Chemie 8:889–896

    Article  Google Scholar 

  • Contestabile R, Paiardini A, Pascarella S, di Salvo ML, D'Aguanno S, Bossa F (2001) L-Threonine aldolase, serine hydroxymethyltransferase and fungal alanine racemase—a subgroup of strictly related enzymes specialized for different functions. Eur J Biochem 268:6508–6525

    Article  CAS  Google Scholar 

  • Coppi L, Giordano C, Longoni A, Panossian S (1997) Thiamphenicol: a manufacturing process involving a double inversion of stereochemistry. In: Collins AN, Sheldrake GN, Crosby J (eds) Chirality in industry, vol 2, 2nd edn. Wiley, Chichester, pp 353–362

    Google Scholar 

  • Dale RA (1978) Catabolism of threonine in mammals by coupling of L-threonine 3-dehydrogenase with 2-amino-3-oxobutyrate-CoA ligase. Biochim Biophys Acta 544:496–503

    CAS  Google Scholar 

  • Dietz FR, Gröger H (2009) Asymmetric synthesis of all stereoisomers of α-methylthreonine using an organocatalytic Steglich rearrangement reaction as a key step. Synthesis-Stuttg 24:4208–4218

    Google Scholar 

  • Edgar AJ (2002) Molecular cloning and tissue distribution of mammalian L-threonine 3-dehydrogenases. BMC Biochem 3:19

    Article  Google Scholar 

  • Edgar AJ (2005) Mice have a transcribed L-threonine aldolase/GLY1 gene, but the human GLY1 gene is a non-processed pseudogene. BMC Genomics 6:32. doi:10.1186/1471-2164-1186-1132

    Article  CAS  Google Scholar 

  • Eisenstein E (1995) Allosteric regulation of biosynthetic threonine deaminase from Escherichia coli: effects of isoleucine and valine on active-site ligand binding and catalysis. Arch Biochem Biophys 316:311–318

    Article  CAS  Google Scholar 

  • Fesko K, Giger L, Hilvert D (2008a) Synthesis of β-hydroxy-α-amino acids with a reengineered alanine racemase. Bioorg Med Chem Lett 18:5987–5990

    Article  CAS  Google Scholar 

  • Fesko K, Reisinger C, Steinreiber J, Weber H, Schurmann M, Griengl H (2008b) Four types of threonine aldolases: similarities and differences in kinetics/thermodynamics. J Mol Catal B Enzym 52–3:19–26

    Article  CAS  Google Scholar 

  • Fesko K, Uhl M, Steinreiber J, Gruber K, Griengl H (2010) Biocatalytic access to α, α-dialkyl-alpha-amino acids by a mechanism-based approach. Angew Chem Int Ed 49:121–124

    CAS  Google Scholar 

  • Fujii M, Miura T, Kajimoto T, Ida Y (2000) Facile synthesis of 3,4-dihydroxyprolines as an application of the L-threonine aldolase-catalyzed aldol reaction. Synlett 2000:1046–1048

    Article  Google Scholar 

  • Garbaccio RM, Wolkenberg SE (2006) Science of synthesis, vol 20b. Georg Thieme Verlag, Stuttgart, pp 1131–1202

    Google Scholar 

  • Gasparski CM, Miller MJ (1991) Synthesis of β-hydroxy-α-amino acids by aldol condensation using a chiral phase-transfer catalyst. Tetrahedron 47:5367–5378

    Article  CAS  Google Scholar 

  • Gijsen H, Qiao L, Fitz W, Wong C-H (1996) Recent advances in the chemoenzymatic synthesis of carbohydrates and carbohydrate mimetics. Chem Rev 96:443–474

    Article  CAS  Google Scholar 

  • Gutierrez LM, Garrabou X, Agosta E, Servi S, Parella T, Joglar J, Clapés P (2008) Serine hydroxymethyl transferase from Streptococcus thermophilus and 2'-L-threonine aldolase from Escherichia coli as stereocomplementary biocatalysts for the synthesis of β-hydroxy-α, ω-diamino acid derivatives. Chem Eur J 14:4647–4656

    Article  CAS  Google Scholar 

  • Gwon H-J, Baik S-H (2010) Diastereoselective synthesis of l-threo-3, 4-dihydroxyphenylserine by low-specific l-threonine aldolase mutants. Biotechnol Lett 32:143–149

    Article  CAS  Google Scholar 

  • Herbert RB, Wilkinson B (1994) Preparation of (2R,3S)-β-hydroxy-α-amino acids by use of a novel Streptomyces aldolase as a resolving agent for racemic material. Can J Chem 72:114–117

    CAS  Google Scholar 

  • Herbert RB, Wilkinson B, Ellames GJ, Kunec EK (1993) Stereospecific lysis of a range of β-hydroxy-α-amino acids catalyzed by a novel aldolase from Streptomyces amakusaensis. Chem Soc Chem Commun 1993:205–206

    Article  Google Scholar 

  • Horikawa M, Busch-Petersen J, Corey EJ (1999) Enantioselective synthesis of β-hydroxy-α-amino acid esters by aldol coupling using a chiral quaternary ammonium salt as catalyst. Tetrahedron Lett 40:3843–3846

    Article  CAS  Google Scholar 

  • Kataoka M, Ikemi M, Morikawa T, Miyoshi T, Nishi K, Wada M, Yamada H, Shimizu S (1997) Isolation and characterization of D-threonine aldolase, a pyridoxal-5'-phosphate-dependent enzyme from Arthrobacter sp. DK-38. Eur J Biochem 248:385–393

    Article  CAS  Google Scholar 

  • Kielkopf CL, Burley SK (2002) X-ray structures of threonine aldolase complexes: structural basis of substrate recognition. Biochemistry 41:11711–11720

    Article  CAS  Google Scholar 

  • Kimura T, Vassilev VP, Shen GJ, Wong CH (1997) Enzymatic synthesis of β-hydroxy-α-amino acids based on recombinant D- and L-threonine aldolases. J Am Chem Soc 119:11734–11742

    Article  CAS  Google Scholar 

  • Kumagai H, Yoshida H, Yamada H, Nagate T (1972) Threonine aldolase from Candida humicola 2. Purification, crystallization and properties. Biochim Biophys Acta 258:779–790

    CAS  Google Scholar 

  • Kuroda S, Nozaki H, Watanabe K, Yokozeki K, Imabayashi Y (2008) Producing L-serine derivative e.g. α-methyl-L-serine which is useful as pharmaceutical intermediate, involves reacting L-alpha-amino acid with aldehyde, in presence of specific enzyme derived from Ralstonia genus. In: Pat Appl EP 1882737A1. Ajinomoto Co Inc.

  • Lee SJ, Kang HY, Lee Y (2003) High-throughput screening methods for selecting L-threonine aldolases with improved activity. J Mol Catal B Enzym 26:265–272

    Article  CAS  Google Scholar 

  • Liu JQ, Dairi T, Kataoka M, Shimizu S, Yamada H (1997) L-allo-threonine aldolase from Aeromonas jandaei DK-39: gene cloning, nucleotide sequencing, and identification of the pyridoxal 5'-phosphate-binding lysine residue by site-directed mutagenesis. J Bacteriol 179:3555–3560

    CAS  Google Scholar 

  • Liu JQ, Ito S, Dairi T, Itoh N, Shimizu S, Yamada H (1998) Low-specificity L-threonine aldolase of Pseudomonas sp. NCIMB 10558: purification, characterization and its application to beta-hydroxy-alpha-amino acid synthesis. Appl Microbiol Biotechnol 49:702–708

    Article  CAS  Google Scholar 

  • Liu JQ, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (1998a) Gene cloning, biochemical characterization and physiological role of a thermostable low-specificity L-threonine aldolase from Escherichia coli. Eur J Biochem 255:220–226

    Article  CAS  Google Scholar 

  • Liu JQ, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (1998b) A novel metal-activated pyridoxal enzyme with a unique primary structure, low specificity D-threonine aldolase from Arthrobacter sp. strain DK-38—molecular cloning and cofactor characterization. J Biol Chem 273:16678–16685

    Article  CAS  Google Scholar 

  • Liu JQ, Ito S, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (1998c) Gene cloning, nucleotide sequencing, and purification and characterization of the low-specificity L-threonine aldolase from Pseudomonas sp. strain NCIMB 10558. Appl Environm Microbiol 64:549–554

    CAS  Google Scholar 

  • Liu JQ, Odani M, Dairi T, Itoh N, Shimizu S, Yamada H (1999) A new route to L-threo-3-[4-(methylthio)phenylserine], a key intermediate for the synthesis of antibiotics: recombinant low-specificity D-threonine aldolase-catalyzed stereospecific resolution. Appl Microbiol Biotechnol 51:586–591

    Article  CAS  Google Scholar 

  • Liu JQ, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (2000a) Diversity of microbial threonine aldolases and their application. J Mol Catal B Enzym 10:107–115

    Article  CAS  Google Scholar 

  • Liu JQ, Odani M, Yasuoka T, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (2000b) Gene cloning and overproduction of low-specificity D-threonine aldolase from Alcaligenes xylosoxidans and its application for production of a key intermediate for parkinsonism drug. Appl Microbiol Biotechnol 54:44–51

    Article  CAS  Google Scholar 

  • Makart S, Bechtold M, Panke S (2007) Towards preparative asymmetric synthesis of β-hydroxy-α-amino acids: L-allo-threonine formation from glycine and acetaldehyde using recombinant GlyA. J Biotechnol 130:402–410

    Article  CAS  Google Scholar 

  • Misono H, Maeda H, Tuda K, Ueshima S, Miyazaki N, Nagata S (2005) Characterization of an inducible phenylserine aldolase from Pseudomonas putida 24-1. Appl Environm Microbiol 71:4602–4609

    Article  CAS  Google Scholar 

  • Miura T, Fujii M, Shingu K, Koshimizu I, Naganoma J, Kajimoto T, Ida Y (1998) Application of L-threonine aldolase-catalyzed reaction for the preparation of a peptidic mimetic of RNA: a leading compound of Vero-toxin inhibitors. Tetrahedron Lett 39:7313–7316

    Article  CAS  Google Scholar 

  • Moon SH, Ohfune Y (1994) Efficient syntheses of the 4 enantiomers and diastereomers of α-methylthreonine and both enantiomers of α-methylserine. J Am Chem Soc 116:7405–7406

    Article  CAS  Google Scholar 

  • Nishiyama T, Mohile SS, Kajimoto T, Node M (2007) Synthesis of thymine polyoxin C by using L-threonine aldolase-catalyzed aldol reaction. Heterocycles 71:1397–1405

    Article  CAS  Google Scholar 

  • Nishiyama T, Kajimoto T, Mohile SS, Hayama N, Otsuda T, Ozeki M, Node M (2009) The first enantioselective synthesis of imino-deoxydigitoxose and protected imino-digitoxose by using L-threonine aldolase-catalyzed aldol condensation. Tetrahedron Asymmetry 30:230–234

    Article  CAS  Google Scholar 

  • Nozaki H, Kuroda S, Watanabe K, Yokozeki K (2008) Gene cloning of α-methylserine aldolase from Variovorax paradoxus and purification and characterization of the recombinant enzyme. Biosci Biotechnol Biochem 72:2580–2588

    Article  CAS  Google Scholar 

  • Nozaki H, Kuroda S, Watanabe K, Yokozeki K (2009) Gene cloning, purification, and characterization of α-methylserine aldolase from Bosea sp AJ110407 and its applicability for the enzymatic synthesis of α-methyl-L-serine and α-ethyl-L-serine. J Mol Catal B Enzym 59:237–242

    Article  CAS  Google Scholar 

  • Ogawa H, Gomi T, Fujioka M (2000) Serine hydroxymethyltransferase and threonine aldolase: are they identical? Int J Biochem Cell Biol 32:289–301

    Article  CAS  Google Scholar 

  • Ooi T, Taniguchi M, Kameda M, Maruoka K (2002) Direct asymmetric aldol reactions of glycine Schiff base with aldehydes catalyzed by chiral quaternary ammonium salts. Angew Chem Int Ed 41:4542–4544

    Article  CAS  Google Scholar 

  • Riario-Sforza G, Pagani R, Marinello E (1969) Threonine aldolase and allothreonine aldolase in rat liver. Eur J Biochem 8:88–92

    Article  CAS  Google Scholar 

  • Sagui F, Conti P, Roda G, Contestabile R, Riva S (2008) Enzymatic synthesis of ω-carboxy-β-hydroxy-(L)-α-amino acids. Tetrahedron 64:5079–5084

    Article  CAS  Google Scholar 

  • Samach A, Hareven D, Gutfinger T, Ken-Dror S, Lifschitz E (1991) Biosynthetic threonine deaminase gene of tomato: isolation, structure, and upregulation in floral organs. Proc Natl Acad Sci USA 88:2678–2682

    Article  CAS  Google Scholar 

  • Schirch L (1982) Serine hydroxymethyltransferase. Adv Enzymol Relat Areas Mol Biol 53:83–112

    CAS  Google Scholar 

  • Schirch LV, Gross T (1968) Serine transhydroxymethylase. J Biol Chem 243:5651–5655

    CAS  Google Scholar 

  • Schöllkopf U, Hartwig W, Groth U (1980) Asymmetric syntheses via heterocyclic intermediates. 3. Enantioselective synthesis of α-methylserines. Angew Chem Int Ed Engl 19:212–213

    Article  Google Scholar 

  • Seebeck FP, Hilvert D (2003) Conversion of a PLP-dependent racemase into an aldolase by a single active site mutation. J Am Chem Soc 125:10158–10159

    Article  CAS  Google Scholar 

  • Shao H, Rueter JK, Goodman M (1998) Novel enantioselective synthesis of α-methylthreonines and α, β-dimethylcysteines. J Org Chem 63:5240–5244

    Article  CAS  Google Scholar 

  • Steinreiber J, Fesko K, Mayer C, Reisinger C, Schürmann M, Griengl H (2007a) Synthesis of γ-halogenated and long-chain β-hydroxy-α-amino acids and 2-amino-1,3-diols using threonine aldolases. Tetrahedron 63:8088–8093

    Article  CAS  Google Scholar 

  • Steinreiber J, Fesko K, Reisinger C, Schürmann M, van Assema F, Wolberg M, Mink D, Griengl H (2007b) Threonine aldolases—an emerging tool for organic synthesis. Tetrahedron 63:918–926

    Article  CAS  Google Scholar 

  • Steinreiber J, Schürmann M, van Assema F, Wolberg M, Fesko K, Reisinger C, Mink D, Griengl H (2007c) Synthesis of aromatic 1, 2-amino alcohols utilizing a bienzymatic dynamic kinetic asymmetric transformation. Adv Synth Catal 349:1379–1386

    Article  CAS  Google Scholar 

  • Steinreiber J, Schürmann M, Wolberg M, van Assema F, Reisinger C, Tesko K, Mink D, Griengl H (2007d) Overcoming thermodynamic and kinetic limitations of aldolase-catalyzed reactions by applying multienzymatic dynamic kinetic asymmetric transformations. Angew Chem Int Ed 46:1624–1626

    Article  CAS  Google Scholar 

  • Stocklein W, Schmidt HL (1985) Evidence for L-threonine cleavage and allo-threonine formation by different enzymes from Clostridium pasteurianum—threonine aldolase and serine hydroxymethyltransferase. Biochem J 232:621–622

    CAS  Google Scholar 

  • Tanaka T, Ozawa M, Miura T, Inazu T, Tsuji S, Kajimoto T (2002) Synthesis of novel mimetics of CMP-sialic acid as the inhibitors of sialyltransferases. Synlett 9:1487–1490

    Google Scholar 

  • Tanaka T, Tsuda C, Miura T, Inazu T, Tsuji S, Nishihara S, Hisamatsu M, Kajimoto T (2004) Design and synthesis of peptide mimetics of GDP-fucose: targeting inhibitors of fucosyltransferases. Synlett 2:243–246

    Google Scholar 

  • Vassilev VP, Uchiyama T, Kajimoto T, Wong CH (1995) L-Threonine aldolase in organic synthesis—preparation of novel β-hydroxy-α-amino acids. Tetrahedron Lett 36:4081–4084

    Article  CAS  Google Scholar 

  • Vidal L, Calveras J, Clapes P, Ferrer P, Caminal G (2005) Recombinant production of serine hydroxymethyl transferase from Streptococcus thermophilus and its preliminary evaluation as a biocatalyst. Appl Microbiol Biotechnol 68:489–497

    Article  CAS  Google Scholar 

  • Wada M, Sakamoto M, Kataoka M, Liu JQ, Yamada H, Shimizu S (1998) Distribution of threonine aldolase activity with different stereospecificities in aerobic bacteria. Biosci Biotechnol Biochem 62:1586–1588

    Article  CAS  Google Scholar 

  • Yamada H, Kumagai H, Nagate T, Yoshida H (1971) Formation of threonine aldolase by bacteria and yeasts. Agric Biol Chem 35:1340–1345

    CAS  Google Scholar 

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Authors and Affiliations

  1. evocatal GmbH, Merowingerplatz 1a, 40225, Düsseldorf, Germany

    Nina Dückers

  2. Department of Chemistry and Pharmacy, University of Erlangen-Nuremberg, Henkestr. 42, 91054, Erlangen, Germany

    Katrin Baer, Sabine Simon & Harald Gröger

  3. Institute of Molecular Enzyme Technology, Heinrich-Heine-University of Düsseldorf, Research Centre Jülich, Stetternicher Forst, 52426, Jülich, Germany

    Werner Hummel

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  1. Nina Dückers
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  2. Katrin Baer
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  4. Harald Gröger
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Correspondence to Harald Gröger or Werner Hummel.

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Dückers, N., Baer, K., Simon, S. et al. Threonine aldolases—screening, properties and applications in the synthesis of non-proteinogenic β-hydroxy-α-amino acids. Appl Microbiol Biotechnol 88, 409–424 (2010). https://doi.org/10.1007/s00253-010-2751-8

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