Skip to main content
SpringerLink
Log in

Overexpression of d-psicose 3-epimerase from Ruminococcus sp. in Escherichia coli and its potential application in d-psicose production

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

The d-psicose 3-epimerase (DPE) gene from Ruminococcus sp. was cloned and overexpressed in Escherichia coli. The recombinant protein was purified and characterized. It was optimally active at pH 7.5–8.0 and 60 °C. Activity was not dependent on the presence of metal ions; however, it became more thermostable with added Mn2+. The K m of the enzyme for d-psicose (48 mM) was lower than that for d-tagatose (230 mM), suggesting that d-psicose is the optimum substrate. More importantly, the thermostability of the novel DPE from Ruminococcus is the strongest among all of the d-psicose and d-tagatose 3-epimerases and may be suitable for the industrial production of d-psicose from fructose.

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

Similar content being viewed by others

References

  • Baek SH, Park SJ, Lee HG (2010) d-Psicose, a sweet monosaccharide, ameliorate hyperglycemia, and dyslipidemia in C57BL/6 J db/db mice. J Food Sci 75:49–53

    Article  Google Scholar 

  • Chan HS, Zhu Y, Hu Y, Ko TP, Huang CH, Ren F, Chen CC, Ma Y, Guo RT, Sun Y (2012) Crystal structures of d-psicose 3-epimerase from Clostridium cellulolyticum H10 and its complex with ketohexose sugars. Protein Cell 3:1–9. doi:10.1007/s13238-012-2026-5

    Article  Google Scholar 

  • Hayashi N, Iida T, Yamada T, Okuma K, Takehara I, Yamamoto T, Yamada K, Tokuda M (2010) Study on the postprandial blood glucose suppression effect of d-psicose in borderline diabetes and the safety of long-term ingestion by normal human subjects. Biosci Biotechnol Biochem 74:510–519

    Article  PubMed  CAS  Google Scholar 

  • Hossain MA, Kitagaki S, Nakano D, Nishiyama A, Funamoto Y, Matsunaga T, Tsukamoto I, Yamaguchi F, Kamitori K, Dong Y, Hirata Y, Murao K, Toyoda Y, Tokuda M (2011) Rare sugar d-psicose improves insulin sensitivity and glucose tolerance in type 2 diabetes Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Biochem Biophys Res Commun 405:7–12

    Article  PubMed  CAS  Google Scholar 

  • Iida T, Hayashi N, Yamada T, Yoshikawa Y, Miyazato S, Kishimoto Y, Okuma K, Tokuda M, Izumori K (2010) Failure of d-psicose absorbed in the small intestine to metabolize into energy and its low large intestinal fermentability in humans. Metabolism 59:206–214

    Article  PubMed  CAS  Google Scholar 

  • Ishida Y, Kamiya T, Itoh H, Kimura Y, Izumori K (1997a) Cloning and characterization of the d-tagatose 3-epimerase gene from Pseudomonas cichorii ST-24. J Ferment Bioeng 83:529–534

    Article  CAS  Google Scholar 

  • Ishida Y, Kamiya T, Izumori K (1997b) Production of d-tagatose 3-epimerase of Pseudomonas cichorii ST-24 using recombinant Escherichia coli. J Ferment Bioeng 84:348–350

    Article  CAS  Google Scholar 

  • Itoh H, Okaya H, Rahman AK, Tajima S, Hayakawa S, Izumori K (1994) Purification and characterization of d-tagatose 3-epimerase from Pseudomonas sp. ST-24. Biosci Biotechnol Biochem 58:2168–2171

    Article  CAS  Google Scholar 

  • Izumori K, Rahman AK, Okaya H, Tsumura T (1993) A new enzyme, d-ketohexose 3-epimerase, from Pseudomonas sp. ST-24. Biosci Biotechnol Biochem 57:1037–1039

    Article  CAS  Google Scholar 

  • Kim HJ, Hyun EK, Kim YS, Lee YJ, Oh DK (2006a) Characterization of an Agrobacterium tumefaciens d-psicose 3-epimerase that converts d-fructose to d-psicose. Appl Environ Microbiol 72:981–985

    Article  PubMed  CAS  Google Scholar 

  • Kim K, Kim HJ, Oh DK, Cha SS, Rhee S (2006b) Crystal structure of d-psicose 3-epimerase from Agrobacterium tumefaciens and its complex with true substrate d-fructose: a pivotal role of metal in catalysis, an active site for the non-phosphorylated substrate, and its conformational changes. J Mol Biol 361:920–931

    Article  PubMed  CAS  Google Scholar 

  • Kim NH, Kim HJ, Kang DI, Jeong KW, Lee JK, Kim Y, Oh DK (2008) Conversion shift of d-fructose to d-psicose for enzyme-catalyzed epimerization by addition of borate. Appl Environ Microbiol 74:3008–3013

    Article  PubMed  CAS  Google Scholar 

  • Mu W, Chu F, Xing Q, Yu S, Zhou L, Jiang B (2011) Cloning, expression, and characterization of a d-psicose 3-epimerase from Clostridium cellulolyticum H10. J Agric Food Chem 59:7785–7792

    Article  PubMed  CAS  Google Scholar 

  • Sakuraba H, Yoneda K, Satomura T, Kawakami R, Ohshima T (2009) Structure of a d-tagatose 3-epimerase-related protein from the hyperthermophilic bacterium Thermotoga maritima. Acta Crystallogr F 65:199–203

    Article  Google Scholar 

  • Takeshita K, Suga A, Takata G, Izumori K (2000) Mass production of d-psicose from d-fructose by a continuous bioreactor system using immobilized d-tagatose 3-epimerase. J Biosci Bioeng 90:453–455

    PubMed  CAS  Google Scholar 

  • Yagi K, Matsuo T (2009) The study on long-term toxicity of d-psicose in rats. J Clin Biochem Nutr 45:271–277

    Article  PubMed  Google Scholar 

  • Zhang L, Mu W, Jiang B, Zhang T (2009) Characterization of d-tagatose-3-epimerase from Rhodobacter sphaeroides that converts d-fructose into d-psicose. Biotechnol Lett 31:857–862

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Innovation Key Program of the Chinese Academy of Sciences (No. KSCX2EWG5), the National High Technology and Research Development Program (No. 2012AA021503), and the Chinese Academy of Sciences Visiting Professorships for Senior International Scientists (No. 2010T1S4).

Author information

Authors and Affiliations

  1. National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Xiqi Road 32#, Tianjin Airport Economic Area, Tianjin, 300308, People’s Republic of China

    Yueming Zhu, Yan Men, Wei Bai, Xiaobo Li, Lili Zhang, Yuanxia Sun & Yanhe Ma

Authors
  1. Yueming Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yan Men
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiaobo Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lili Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuanxia Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yanhe Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yuanxia Sun.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 246 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhu, Y., Men, Y., Bai, W. et al. Overexpression of d-psicose 3-epimerase from Ruminococcus sp. in Escherichia coli and its potential application in d-psicose production. Biotechnol Lett 34, 1901–1906 (2012). https://doi.org/10.1007/s10529-012-0986-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-012-0986-4

Keywords

Search

Navigation