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Preparation of optically active β-hydroxy-α-amino acid by immobilized Escherichia coli cells with serine hydroxymethyl transferase activity

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Abstract

In this research, an improved method for preparation of optically pure β-hydroxy-α-amino acids, catalyzed by serine hydroxymethyl transferase with threonine aldolase activity, is reported. Using recombinant serine hydroxymethyl transferase (SHMT), an enzymatic resolution process was established. A series of new substrates, β-phenylserine, β-(nitrophenyl) serine and β-(methylsulfonylphenyl) serine were used in the resolution process catalyzed by immobilized Escherichia coli cells with SHMT activity. It was observed that the K m for l-threonine was 28-fold higher than that for l-allo-threonine, suggesting that this enzyme can be classified as a low-specificity l-allo-threonine aldolase. The results also shows that SHMT activity with β-phenylserine as substrate was about 1.48-fold and 1.25-fold higher than that with β-(methylsulfonylphenyl) serine and β-(nitrophenyl) serine as substrate, respectively. Reaction conditions were optimized by using 200 mmol/l β-hydroxy-α-amino acid, and 0.1 g/ml of immobilized SHMT cells at pH 7.5 and 45°C. Under these conditions, the immobilized cells were continuously used 10 times, yielding an average conversion rate of 60.4%. Bead activity did not change significantly the first five times they were used, and the average conversion rate during the first five instances was 84.1%. The immobilized cells exhibited favourable operational stability.

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References

  • Apley M (1997a) Ancillary therapy of bovine respiratory disease. Veterinary Clinics of North America-Food Animal Practice 13:575–582

    CAS  Google Scholar 

  • Apley M (1997b) Antimicrobial therapy of bovine respiratory disease. Veterinary Clinics of North America-Food Animal Practice 13:549–556

    CAS  Google Scholar 

  • Birgisson H, Wheat JO, Hreggvidsson GO, Kristjansson JK, Mattiasson B (2007) Immobilization of a recombinant Escherichia coli producing a thermostable α-l-rhamnosidase: Creation of a bioreactor for hydrolyses of naringin. Enzyme Microb Tech 40:1181–1187

    Article  CAS  Google Scholar 

  • Burke TR, Knight M, Chandrasekhar B (1989) Solid-phase synthesis of viscosin, a cyclic depsipeptide with antibacterial and antiviral properties. Tetrahedron Lett 30:519–522

    Article  CAS  Google Scholar 

  • Fukushima T, Santa T, Homma H, Nagatomo R, Imai K (1995) Determination of d-amino acids in serum from patients with renal dysfunction. Biol Pharm Bull 18:1130–1132

    CAS  PubMed  Google Scholar 

  • Hayashi H (1995) Pyridoxal enzymes-mechanistic diversity and uniformity. J Biochem 118:463–473

    CAS  PubMed  Google Scholar 

  • Kajimura Y, Kaneda M (1996) Fusaricidin A, a new depsipeptide antibiotic produced by Bacillus polymyxa KT-8-Taxonomy, fermentation, isolation, structure elucidation and biological activity. J Antibiot 49:129–135

    CAS  PubMed  Google Scholar 

  • Koch, Melvin V, Magni, Ambrogio (1985) Process for the production of serine derivatives. US Patent 4,501, 919

  • 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  PubMed  Google Scholar 

  • Maruyama W, Naoi M, Narabayashi H (1996) The metabolism of l-DOPA and l-threo-3, 4-dihydroxyphenylserine and their effects on monoamines in the human brain: analysis of the intraventricular fluid from parkinsonian patients. J Neurol Sci 139:141–148

    Article  CAS  PubMed  Google Scholar 

  • Matthews RG, Drummond JT (1990) Providing one-carbon units for biological methylations-mechanistic studies on serine hydroxymethyltransferase, methylenetetrahydrofolate reductase, and methyltetrahydrofolate-homocysteine methyltransferase. Chem Rev 90:1275–1290

    Article  CAS  Google Scholar 

  • Mehta PK, Christen P (2000) The molecular evolution of pyridoxal-5′-phosphate-dependent enzymes. Adv Enzymol 74:129–137

    CAS  PubMed  Google Scholar 

  • Miyazaki SS, Toki S, Izumi Y, Yamada H (1987) Further characterization of serine hydroxymethyltransferase from a serine-producing methylotroph, hyphomicrobium-methylovorum. Agric Biol Chem 51:2587–2589

    CAS  Google Scholar 

  • Reetz MT (2001) Combinatorial and evolution-based methods in the creation of enantioselective catalysts. Angew Chem Int Edit 40:284–310

    Article  CAS  Google Scholar 

  • Renwick SB, Snell K, Baumann U (1998) The crystal structure of human cytosolic serine hydroxymethyltransferase: a target for cancer chemotherapy. Structure 6:1105–1116

    Article  CAS  PubMed  Google Scholar 

  • Scarsdale JN, Kazanina G, Radaev S, Schirch V, Wright HT (1999) Crystal structure of rabbit cytosolic serine hydroxymethyltransferase at 2.8 angstrom resolution: mechanistic implications. Biochem 38:8347–8358

    Article  CAS  Google Scholar 

  • Scarsdale JN, Radaev S, Kazanina G, Schirch V, Wright HT (2000) Crystal structure at 2.4 angstrom resolution of E. coli serine hydroxymethyltransferase in complex with glycine substrate and 5-formyl tetrahydrofolate. J Mol Biol 296:155–168

    Article  CAS  PubMed  Google Scholar 

  • Schirch L, Gross T (1968) Serine transhydroxymethylase-identification as threonine and allothreonine aldolases. J Biol Chem 243:5651–5657

    CAS  PubMed  Google Scholar 

  • Schirch V, Hopkins S, Villar E, Angelaccio S (1985) Serine hydroxymethyltransferase from Escherichia coli-purification and properties. J Bacteriol 163:1–7

    CAS  PubMed  Google Scholar 

  • Shiraiwa T, Saijoh R, Suzuki M, Yoshida K, Nishimura S, Nagasawa H (2003) Preparation of optically active threo-2-amino-3-hydroxy-3-phenylpropanoic acid (threo-β-phenylserine) via optical resolution. Chem Pharm Bull 51:1363–1367

    Article  CAS  PubMed  Google Scholar 

  • Shostak K, Schirch V (1988) Serine hydroxymethyltransferase-mechanism of the racemization and transamination of d-alanine and l-alanine. Biochem 27:8007–8014

    Article  CAS  Google Scholar 

  • Soloshonok VA, Hayashi T (1994a) Gold(I)-catalyzed asymmetric aldol reaction of methyl isocyanoacetate with fluorinated benzaldehydes. Tetrahedron Lett 35:2713–2716

    Article  CAS  Google Scholar 

  • Soloshonok VA, Hayashi T (1994b) Gold(I)-catalyzed asymmetric aldol reactions of fluorinated benzaldehydes with an α-isocyanoacetamide. Tetrahedron-Asymmetry 5:1091–1094

    Article  CAS  Google Scholar 

  • Soloshonok VA, Belokon YN, Kuzmina NA, Maleev VI, Svistunova NY, Solodenko VA, Kukhar VP (1992) Asymmetric-synthesis of phosphorus analogs of dicarboxylic α-amino-acids. J Chem Soc Perk T 1:1525–1529

    Google Scholar 

  • Soloshonok VA, Kukhar VP, Galushko SV, Svistunova NY, Avilov DV, Kuzmina NA, Raevski NI, Struchkov YT, Pysarevsky AP, Belokon YN (1993) General-method for the synthesis of enantiomerically pure β-hydroxy-α-amino acids, containing fluorine-atoms in the side-chains-case of stereochemical distinction between methyl and trifluoromethyl groups. X-ray crystal and molecular-structure of the nickel(II) complex of (2S, 3S)-2-(trifluoromethyl)threonine. J Chem Soc Perk T 1:3143–3155

    Google Scholar 

  • Soloshonok VA, Hayashi T, Ishikawa K, Nagashima N (1994c) Highly diastereoselective aldol reaction of fluoroalkyl aryl ketones with methyl isocyanoacetate catalyzed by silver(I)/triethylamine. Tetrahedron Lett 35:1055–1058

    Article  CAS  Google Scholar 

  • Soloshonok VA, Kacharov AD, Avilov DV, Hayashi T (1996a) Transition metal-catalyzed diastereoselective aldol reactions of prochiral ketones with methyl isocyanoacetate. Tetrahedron Lett 37:7845–7848

    Article  CAS  Google Scholar 

  • Soloshonok VA, Kacharov AD, Hayashi T (1996b) Gold(I)-catalyzed asymmetric aldol reactions of isocyanoacetic acid derivatives with fluoroaryl aldehydes. Tetrahedron 52:245–254

    Article  CAS  Google Scholar 

  • Soloshonok VA, Kacharov AD, Avilov DV, Ishikawa K, Nagashima N, Hayashi T (1997) Transition metal/base-catalyzed aldol reactions of isocyanoacetic acid derivatives with prochiral ketones, a straightforward approach to stereochemically defined β, β-disubstituted-β-hydroxy-α-amino acids. Scope and limitations. J Org Chem 62:3470–3479

    Article  CAS  Google Scholar 

  • Straathof AJJ, Panke S, Schmid A (2002) The production of fine chemicals by biotransformations. Curr Opin Biotech 13:548–556

    Article  CAS  PubMed  Google Scholar 

  • Szebenyi DME, Liu XW, Kriksunov IA, Stover PJ, Thiel DJ (2000) Structure of a murine cytoplasmic serine hydroxymethyltransferase quinonoid ternary complex: evidence for asymmetric obligate dimers. Biochem 39:13313–13323

    Article  CAS  Google Scholar 

  • Szebenyi DME, Musayev FN, di Salvo ML, Safo MK, Schirch V (2004) Serine hydroxymethyltransferase: Role of Glu75 and evidence that serine is cleaved by a retroaldol mechanism. Biochem 43:6865–6876

    Article  CAS  Google Scholar 

  • Ulevitch RJ, Kallen RG (1977) Purification and characterization of pyridoxal-5’-phosphate dependent serine hydroxymethylase from lamb liver and its action upon β-phenylserines. Biochemistry 16:5342–5350

    Article  CAS  PubMed  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 Biot 68:489–497

    Article  CAS  Google Scholar 

  • Zheng RC, Zheng YG, Shen YC (2007) A simple method to determine concentration of enantiomers in enzyme-catalyzed kinetic resolution. Biotechnol Lett 29:1087–1091

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Technology-Innovation Fund (02CJ-13-01-16) and the Open Fund of State Key Laboratory of Pharmaceutical Biotechnology of Nanjing University, P.R. China.

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  1. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing, 210093, People’s Republic of China

    Gen-Hai Zhao, Hui Li, Wei Liu, Wei-Guo Zhang, Fei Zhang, Qian Liu & Qing-Cai Jiao

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  1. Gen-Hai Zhao
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  2. Hui Li
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  3. Wei Liu
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  4. Wei-Guo Zhang
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  5. Fei Zhang
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  7. Qing-Cai Jiao
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Correspondence to Qing-Cai Jiao.

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Zhao, GH., Li, H., Liu, W. et al. Preparation of optically active β-hydroxy-α-amino acid by immobilized Escherichia coli cells with serine hydroxymethyl transferase activity. Amino Acids 40, 215–220 (2011). https://doi.org/10.1007/s00726-010-0637-9

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  • DOI: https://doi.org/10.1007/s00726-010-0637-9

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