Applied Microbiology and Biotechnology

, Volume 99, Issue 19, pp 7963–7972 | Cite as

Biosynthesis of 2-deoxysugars using whole-cell catalyst expressing 2-deoxy-d-ribose 5-phosphate aldolase

  • Jitao Li
  • Jiangang Yang
  • Yan Men
  • Yan Zeng
  • Yueming Zhu
  • Caixia Dong
  • Yuanxia SunEmail author
  • Yanhe Ma
Biotechnological products and process engineering


2-Deoxy-d-ribose 5-phosphate aldolase (DERA) accepts a wide variety of aldehydes and is used in de novo synthesis of 2-deoxysugars, which have important applications in drug manufacturing. However, DERA has low preference for non-phosphorylated substrates. In this study, DERA from Klebsiella pneumoniae (KDERA) was mutated to increase its enzyme activity and substrate tolerance towards non-phosphorylated polyhydroxy aldehyde. Mutant KDERAK12 (S238D/F200I/ΔY259) showed a 3.15-fold improvement in enzyme activity and a 1.54-fold increase in substrate tolerance towards d-glyceraldehyde compared with the wild type. Furthermore, a whole-cell transformation strategy using resting cells of the BL21(pKDERA12) strain, containing the expressed plasmid pKDERA12, resulted in increase in 2-deoxy-d-ribose yield from 0.41 mol/mol d-glyceraldehyde to 0.81 mol/mol d-glyceraldehyde and higher substrate tolerance from 0.5 to 3 M compared to in vitro assays. With further optimization of the transformation process, the BL21(pKDERA12) strain produced 2.14 M (287.06 g/L) 2-deoxy-d-robose (DR), with a yield of 0.71 mol/mol d-glyceraldehyde and average productivity of 0.13 mol/L·h (17.94 g/L·h). These results demonstrate the potential for large-scale production of 2-deoxy-d-ribose using the BL21(pKDERA12) strain. Furthermore, the BL21(pKDERA12) strain also exhibited the ability to efficiently produce 2-deoxy-d-altrose from d-erythrose, as well as 2-deoxy-l-xylose and 2-deoxy-l-ribose from l-glyceraldehyde.


2-Deoxy-d-ribose 5-phosphate aldolase (DERA) Deoxysugars Whole-cell transformation Directed mutagenesis 



This work was supported by the National High Technology Research and Development Program of China (No. 2012AA021403) and Science and Technology Projects of Tianjin (No. 13ZCZDSY05600).

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

253_2015_6740_MOESM1_ESM.pdf (945 kb)
ESM 1 (PDF 945 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Jitao Li
    • 1
  • Jiangang Yang
    • 1
  • Yan Men
    • 1
  • Yan Zeng
    • 1
  • Yueming Zhu
    • 1
  • Caixia Dong
    • 2
  • Yuanxia Sun
    • 1
    Email author
  • Yanhe Ma
    • 1
  1. 1.National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjinChina
  2. 2.Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnosis, School of PharmacyTianjin Medical UniversityTianjinChina

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