Skip to main content

Advertisement

SpringerLink
Log in

Characterization of a serine hydroxymethyltransferase for l-serine enzymatic production from Pseudomonas plecoglossicida

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Pseudomonas plecoglossicida, a bacterium strain that exhibits high Serine hydroxymethyltransferase (SHMT) activity, was isolated from the seawater. A full-length glyA encoding SHMT was obtained by a modified thermal asymmetric interlaced-PCR (TRIL-PCR), which consisted of 1,254 bp, encoded a 417 amino acid polypeptide, and shared the highest identity (75 %) with a glyA gene from Acinetobacter radioresistens CMC-1. Recombinant glyA gene was expressed in Escherichia coli BL21 (DE3) and purified by electrophoretic homogeneity. The enzyme showed the optimal activity at pH 8.0 and 40 °C, and remained stable in high alkali conditions. Using SHMT to produce l-serine by catalyzing the reaction of glycine and tetrahydrofolate is one of the most promising routes to synthesize l-serine, achieving 33.4 mM l-serine at the 12th h of the enzymatic reaction with the substrates of glycine (133 mM) and formaldehyde (13.3 mM). The properties make the SHMT a candidate for further enzymatic studies and industrial applications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Barra D, Martini F, Angelaccio S, Bossa F, Gavilanes F, Peterson D, Bullis B, Schirch L (1983) Sequence homology between prokaryotic and eukaryotic forms of serine hydroxymethyltransferase. Biochem Biophys Res Commun 116(3):1007–1012

    Article  CAS  Google Scholar 

  • Blakley RL (1960) A spectrophotometric study of the reaction catalysed by serine transhydroxymethylase. Biochem J 77(3):459

    CAS  Google Scholar 

  • Blakley RL (1969) The biochemistry of folic acid and related pteridines. North-Holland Publishing Co, Amsterdam

    Google Scholar 

  • Carbone A, Zinovyev A, Kepes F (2003) Codon adaptation index as a measure of dominating codon bias. Bioinformatics 19(16):2005–2015

    Article  CAS  Google Scholar 

  • LU Fa, Z.G.W.-g (2005). Screening of l-Serine Producer and the Fermentation Conditions in Shake-Flask Culture. Journal of Food Science and Biotechnology, 24(2)

  • Garrow T, Brenner A, Whitehead V, Chen XN, Duncan R, Korenberg J, Shane B (1993) Cloning of human cDNAs encoding mitochondrial and cytosolic serine hydroxymethyltransferases and chromosomal localization. J Biol Chem 268(16):11910–11916

    CAS  Google Scholar 

  • Hagishita T, Yoshida T, Izumi Y, Mitsunaga T (1996) Efficient l-serine production from methanol and glycine by resting cells of Methylobacterium sp. strain MN43. Biosci Biotechnol Biochem 60(10):1604–1607

    Article  CAS  Google Scholar 

  • Hamilton B, Hsiao H, Swann W, Anderson D, Delente J (1985) Manufacture of l-amino acids with bioreactors. Trends Biotechnol 3(3):64–68

    Article  CAS  Google Scholar 

  • Hsiao HY, Wei T (1985) Enzymatic production of l-serine with a feedback control system for formaldehyde addition. Biotechnol Bioeng 28(10):1510–1518

    Article  Google Scholar 

  • Izumi Y, Yoshida T, Miyazaki SS, Mitsunaga T, Ohshiro T, Shimao M, Miyata A, Tanabe T (1993) l-Serine production by a methylotroph and its related enzymes. Appl Microbiol Biotechnol 39(4):427–432

    Article  CAS  Google Scholar 

  • John G, Morita M (1999) Synthesis and characterization of photo-cross-linked networks based on l-lactide/serine copolymers. Macromolecules 32(6):1853–1858

    Article  CAS  Google Scholar 

  • Jones C, Priest D (1976) Metabolism of glutamine and ammonia in rat liver: the effects of N-acetylglutamate and phosphate. Arch Biochem Biophys 174:305–311

    Article  CAS  Google Scholar 

  • Kamphuis J, Boesten WHJ, Broxterman QB, Hermes HFM, van Balken JAM, Meijer EM, Schoemaker HE (1990) New developments in the chemo-enzymatic production of amino acids. In: Bioprocesses and Applied Enzymology. Springer, Berlin, pp 133–186

  • Liu YG, Whittier RF (1995) Thermal asymmetric interlaced PCR: automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking. Genomics 25(3):674–681

    Article  CAS  Google Scholar 

  • Lou X, JIA X-j, LIU S-l, CHEN L-h (2009) Optimization of the fermentation medium for mycophenoilc acid production with response surface methodology. Hebei Chem Eng Ind 9:003

    Google Scholar 

  • Maniatis T, Fritsch E, Sambrook. J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  • Mazars GR, Theillet C (1991) Direct sequencing by thermal asymmetric PCR. Methods Mol Biol-Clifton Then Totowa 65:35–40

    Google Scholar 

  • McGavin M, Forsberg C, Crosby B, Bell A, Dignard D, Thomas D (1989) Structure of the cel-3 gene from Fibrobacter succinogenes S85 and characteristics of the encoded gene product, endoglucanase 3. J Bacteriol 171(10):5587–5595

    CAS  Google Scholar 

  • Miyata A, Yoshida T, Yamaguchi K, Yokoyama C, Tanabe T, Toh H, Mitsunaga T, Izumi Y (1993) Molecular cloning and expression of the gene for serine hydroxymethyltransferase from an obligate methylotroph Hyphomicrobium methylovorum GM2. Eur J Biochem 212(3):745–750

    Article  CAS  Google Scholar 

  • Miyazaki SS, Toki S, Izumi Y, Yamada H (1987) Purification and characterization of a serine hydroxymethyltransferase from an obligate methylotroph, Hyphomicrobium methylovorum GM2. Eur J Biochem 162(3):533–540

    Article  CAS  Google Scholar 

  • Oppenheimer CH (1952) The growth and viability of sixty three species of marine bacteria as influenced by hydrostatic pressure. J Mar Res 11:10–18

    Google Scholar 

  • Park SC, Shimamura I, Fukunaga M, Mori K-I, Nakai T (2000) Isolation of bacteriophages specific to a fish pathogen, Pseudomonas plecoglossicida, as a candidate for disease control. Appl Environ Microbiol 66(4):1416–1422

    Article  CAS  Google Scholar 

  • Patt T, Cole G, Hanson R (1976) Methylobacterium, a new genus of facultatively methylotrophic bacteria. Int J Syst Evolut Microbiol 26(2):226

    CAS  Google Scholar 

  • Peters-Wendisch P, Stolz M, Etterich H, Kennerknecht N, Sahm H, Eggeling L (2005) Metabolic engineering of Corynebacterium glutamicum for l-serine production. Appl Environ Microbiol 71(11):7139–7144

    Article  CAS  Google Scholar 

  • Primers P (2007) High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences. Biotechniques 43:649–656

    Article  Google Scholar 

  • Qiao Y, Huang S, Fan X, Zhao W, Wu B (2003) Cloning of the glyA gene encoding the serine hydroxymethyltransferase. Guangxi nongye shengwu kexue J Agric Biol Sci 22(2):139

    Google Scholar 

  • Rao DN, Appaji Rao N (1982) Purification and regulatory properties of mung bean (Vigna radiata L.) serine hydroxymethyltransferase. Plant Physiol 69:11–18

    Article  CAS  Google Scholar 

  • Rose TM, Schultz ER, Henikoff JG, Pietrokovski S, McCallum CM, Henikoff S (1998) Consensus-degenerate hybrid oligonucleotide primers for amplification of distantly related sequences. Nucleic Acids Res 26(7):1628

    Article  CAS  Google Scholar 

  • Rose TM, Henikoff JG, Henikoff S (2003) CODEHOP (COnsensus-DEgenerate hybrid oligonucleotide primer) PCR primer design. Nucleic Acids Res 31(13):3763

    Article  CAS  Google Scholar 

  • Scarsdale JN, Radaev S, Kazanina G, Schirch V, Wright H (2000) Crystal structure at 2.4 Å resolution of E. coli serine hydroxymethyltransferase in complex with glycine substrate and 5-formyl tetrahydrofolate. J Mol Biol 296(1):155–168

    Article  CAS  Google Scholar 

  • Schirch L, Diller A (1971) Serine transhydroxymethylase. Affinity of the active site for substrates, substrate analogues, and anions. J Biol Chem 246(12):3961

    CAS  Google Scholar 

  • Shen PH, Chao HJ, Jiang CJ, Long ZD, Wang CH, Wu B (2010) Enhancing production of l-Serine by increasing the glyA gene expression in methylobacterium sp MB200. Appl Biochem Biotechnol 160(3):740–750

    Article  CAS  Google Scholar 

  • Strege M, Schmidt D, Kreuzman A, Dotzlaf J, Yeh W, Kaiser R, Lagu A (1994) Capillary electrophoretic analysis of serine hydroxymethyltransferase in Escherichia coli fermentation broth. Anal Biochem 223(2):198–204

    Article  CAS  Google Scholar 

  • Ulevitch RJ, Kallen RG (1977) Studies of the reactions of lamb liver serine hydroxymethylase with l-phenylalanine: kinetic isotope effects upon quinonoid intermediate formation. Biochemistry 16(24):5350–5354

    Article  CAS  Google Scholar 

  • Usha R, Savithri HS, Appaji Rao N (1994) The primary structure of sheep liver cytosolic serine hydroxymethyltransferase and an analysis of the evolutionary relationships among serine hydroxymethyltransferases. Biochim Biophys Acta (BBA) Protein Struct Mol Enzymol 1204(1):75–83

    Article  CAS  Google Scholar 

  • Watanabe M, Morinaga Y, Enei H (1987) l-serine production by a methionine-auxotrophic mutant of methylotrophic Pseudomonas. J Ferment Technol 65(6):617–620

    Article  CAS  Google Scholar 

  • Wei X, Kuhn DN, Narasimhan G (2003) Degenerate primer design via clustering. Proc IEEE Comput Soc Bioinform Conf 2:75–83

    Google Scholar 

  • Yamada H, Miyazaki S, Izumi Y (1986) l-serine production by a glycine-resistant mutant of methylotrophic Hyphomicrobium methylovorum. Agric Biol Chem 50(1):17–21

    Article  CAS  Google Scholar 

  • Zhao M, Ma Y, Dai L, Zhang D, Li J, Yuan W, Li Y, Zhou H (2013) A high-performance liquid chromatographic method for simultaneous determination of 21 Free Amino Acids in Tea. Food Anal Methods 6(1):69–75

    Google Scholar 

  • Zuo Z-Y, Zheng Z-L, Liu Z-G, Yi Q-M, Zou G-L (2007) Cloning, DNA shuffling and expression of serine hydroxymethyltransferase gene from Escherichia coli strain AB90054. Enzyme Microb Technol 40(4):569–577

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from China National Natural Sciences Foundation (u1170303).

Author information

Authors and Affiliations

  1. State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Wei Jiang, Bingzhao Xia, Junjie Huang & Ziduo Liu

Authors
  1. Wei Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bingzhao Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Junjie Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ziduo Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ziduo Liu.

Additional information

Wei Jiang and Bingzhao Xia are contributed equally to the work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jiang, W., Xia, B., Huang, J. et al. Characterization of a serine hydroxymethyltransferase for l-serine enzymatic production from Pseudomonas plecoglossicida . World J Microbiol Biotechnol 29, 2067–2076 (2013). https://doi.org/10.1007/s11274-013-1370-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-013-1370-9

Keywords

Search

Navigation