Skip to main content
SpringerLink
Log in

Alanine racemase from the green alga Chlamydomonas reinhardtii

  • Published:
Amino Acids Aims and scope Submit manuscript

Summary.

Chlamydomonas reinhardtii, a unicellular green microalga, could grow to a stationary phase having optical density of 2.0–2.5 at 750 nm in Tris-acetate-phosphate (TAP) medium containing 0.1% D-alanine. D-alanine has no inhibitory effect on growth and induced alanine racemase activity 130-fold more than without D-alanine in the green alga. Although C. reinhardtii cultured in the TAP medium showed alanine racemase activity, the content of free D-alanine was only 0.14%. The enzyme was partially purified by ammonium sulfate fractionation followed by three kinds of liquid chromatography using DEAE Toyopearl, Phenyl Sepharose, and TSK G3000 SWXL columns. The specific activity for L-alanine of the partially purified alanine racemase was 3.8 µmol/min/mg. The molecular weight of the enzyme was determined to be approximately 72,000 by gel filtration. The enzyme showed a maximum activity at 45 °C and pH 8.4 and requires pyridoxal 5′-phosphate as a coenzyme.

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

Similar content being viewed by others

References

  • O Erikson M Hertzberg T Näsholm (2004) ArticleTitleA conditional marker gene allowing both positive and negative selection in plants Nat Biotechnol 22 455–458 Occurrence Handle15085802 Occurrence Handle1:CAS:528:DC%2BD2cXis1yjtr0%3D Occurrence Handle10.1038/nbt946

    Article  PubMed  CAS  Google Scholar 

  • TV Kopytina NI Rekoslavskaya RK Salyaev (1998) ArticleTitleOsmotically regulated tryptophan racemase from plants Dokl Akad Nauk 363 714–718 Occurrence Handle1:CAS:528:DyaK1MXmsVeisrw%3D

    CAS  Google Scholar 

  • OH Lowry NJ Rosebrough AL Farr RJ Randall (1951) ArticleTitleProtein measurement with the Folin phenol reagent J Biol Chem 193 265–275 Occurrence Handle14907713 Occurrence Handle1:CAS:528:DyaG38XhsVyrsw%3D%3D

    PubMed  CAS  Google Scholar 

  • E Nagahisa N Kan-no M Sato Y Sato (1995) ArticleTitleOccurrence of free D-alanine in marine macroalgae Biosci Biotech Biochem 59 2176–2177 Occurrence Handle1:CAS:528:DyaK2MXpslyht74%3D Occurrence Handle10.1271/bbb.59.2176

    Article  CAS  Google Scholar 

  • Y Nagata T Shimojo T Akino (1988) ArticleTitleTwo spectrophotometric assays for D-amino acid oxidase: for the study of distribution patterns Int J Biochem 20 1235–1238 Occurrence Handle2907883 Occurrence Handle1:CAS:528:DyaL1MXntFGgtQ%3D%3D Occurrence Handle10.1016/0020-711X(88)90225-X

    Article  PubMed  CAS  Google Scholar 

  • Y Nagata K Yamamoto T Shimojo (1992) ArticleTitleDetermination of D- and L-amino acids in mouse kidney by high-performance liquid chromatography J Chromatogr 575 147–152 Occurrence Handle1517292 Occurrence Handle1:CAS:528:DyaK38XitFSmsrg%3D

    PubMed  CAS  Google Scholar 

  • NI Rekoslavskaya OV Yur’va LA Shibanova RK Salyaev (1997) ArticleTitleSynthesis and physiological function of D-tryptophan during wheat germination Russ J Plant Physiol 44 196–203 Occurrence Handle1:CAS:528:DyaK2sXit1CqtLs%3D

    CAS  Google Scholar 

  • T Uo T Yoshimura N Tanaka K Takegawa N Esaki (2001) ArticleTitleFunctional characterization of alanine racemase from Schizosaccharomyces pombe: a Eucaryotic counterpart to bacterial alanine racemase J Bacteriol 183 2226–2233 Occurrence Handle11244061 Occurrence Handle1:CAS:528:DC%2BD3MXitFWkurY%3D Occurrence Handle10.1128/JB.183.7.2226-2233.2001

    Article  PubMed  CAS  Google Scholar 

  • T Yokoyama N Kan-no T Ogata Y Kotaki M Sato E Nagahisa (2003) ArticleTitlePresence of free D-amino acids in microalgae Biosci Biotech Biochem 67 388–392 Occurrence Handle1:CAS:528:DC%2BD3sXhvFemtr8%3D Occurrence Handle10.1271/bbb.67.388

    Article  CAS  Google Scholar 

  • T Yokoyama Y Tanaka M Sato N Kan-no T Nakano T Yamaguchi E Nagahisa (2005) ArticleTitleAlanine racemase activity in the microalga Thalassiosira sp Fish Sci 71 924–930 Occurrence Handle1:CAS:528:DC%2BD2MXps1Kqu78%3D Occurrence Handle10.1111/j.1444-2906.2005.01046.x

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Applied Chemistry, Junior College, Nihon University, Chiba, Japan

    K. Nishimura

  2. Department of Materials and Applied Chemistry, College of Science and Technology, Nihon University, Tokyo, Japan

    K. Nishimura, Y. Tomoda, Y. Nakamoto, T. Kawada, Y. Ishii & Y. Nagata

Authors
  1. K. Nishimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y. Tomoda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. Nakamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. Kawada
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Y. Ishii
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y. Nagata
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nishimura, K., Tomoda, Y., Nakamoto, Y. et al. Alanine racemase from the green alga Chlamydomonas reinhardtii . Amino Acids 32, 59–62 (2007). https://doi.org/10.1007/s00726-006-0352-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-006-0352-8

Search

Navigation