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Simultaneous biosynthesis of (

Xiaojing Jia1,2, Robert M Kelly3, Yejun Han1

  • 1National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.

Metabolic Engineering Communications
|September 11, 2018
PubMed
Summary
This summary is machine-generated.

A novel cell-free system efficiently produces pure (R)-acetoin and ethylene glycol (EG) from D-xylose. This ATP-free biocatalytic platform avoids by-products, offering a sustainable route for valuable biomolecules.

Keywords:
(R)-acetoinBSA, bovine serum albuminCofactor regenerationD-xyloseEG, ethylene glycolEMP, Embden-Meyerhoff-ParnasEthylene glycolFAD, flavin adenine dinucleotideGC, gas chromatographyHPLC, high-pressure liquid chromatographyIPTG, isopropyl-β-D-thiogalactopyranosideIn vitro metabolic engineeringLB, lysogeny brothNAD+, oxidized nicotinamide adenine dinucleotideNADH, reduced nicotinamide adenine dinucleotidePET, polyethylene terephthalatePP, pentose phosphateSDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresisThDP, Thiamine diphosphateee, enantiomeric excess

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Area of Science:

  • Biotechnology and Synthetic Biology
  • Biocatalysis and Enzyme Engineering
  • Sustainable Chemistry

Background:

  • Microbial fermentation for (R)-acetoin and ethylene glycol (EG) often yields unwanted by-products.
  • (R)-acetoin is a key precursor for optically active materials, while EG is a high-volume commodity chemical.
  • Current production methods face challenges with purity and efficiency.

Purpose of the Study:

  • To develop a cell-free bioreaction system for co-producing enantiomerically pure (R)-acetoin and EG from biomass-derived D-xylose.
  • To optimize an in vitro biocatalytic pathway that avoids the limitations of traditional fermentation.
  • To demonstrate a novel platform for generating multiple value-added biomolecules from sugars.

Main Methods:

  • A seven-step, ATP-free cell-free system was designed using enzymes from E. coli, B. subtilis, and C. crescentus.
  • In situ cofactor regeneration was incorporated to sustain the biocatalytic cascade.
  • Optimized reaction conditions (temperature, pH) were determined for maximal product yield and purity.

Main Results:

  • Enantiomerically pure (R)-acetoin was produced at 3.2 mM with 99.5% stereoisomeric purity from 10 mM D-xylose.
  • Initial (R)-acetoin productivity reached 1.0 mM/h under optimized conditions (30°C, pH 7.5, 24h).
  • Ethylene glycol (EG) was co-produced at 5.5 mM with an initial productivity of 1.7 mM/h.

Conclusions:

  • A cell-free bioreaction scheme successfully co-produces high-purity (R)-acetoin and EG from D-xylose.
  • The developed ATP-free biocatalytic platform offers a promising alternative to conventional fermentation, minimizing by-product formation.
  • This approach highlights the potential for efficient, sustainable production of multiple valuable chemicals from biomass sugars.