Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Enhancement of 1,3-Dihydroxyacetone Production from Gluconobacter oxydans by Combined Mutagenesis

  • Lin, Xi (School of Life Science and Technology, China Pharmaceutical University) ;
  • Liu, Sha (School of Life Science and Technology, China Pharmaceutical University) ;
  • Xie, Guangrong (School of Life Science and Technology, China Pharmaceutical University) ;
  • Chen, Jing (School of Life Science and Technology, China Pharmaceutical University) ;
  • Li, Penghua (School of Life Science and Technology, China Pharmaceutical University) ;
  • Chen, Jianhua (School of Life Science and Technology, China Pharmaceutical University)
  • Received : 2016.04.07
  • Accepted : 2016.07.20
  • Published : 2016.11.28
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Abstract

Wild strain L-6 was subjected to combined mutagenesis, including UV irradiation, atmospheric and room temperature plasma, and ion beam implantation, to increase the yield of 1,3-dihydroxyacetone (DHA). With application of a high-throughput screening method, mutant Gluconobacter oxydans I-2-239 with a DHA productivity of 103.5 g/l in flask-shake fermentation was finally obtained with the starting glycerol concentration of 120 g/l, which was 115.7% higher than the wild strain. The cultivation time also decreased from 54 h to 36 h. Compared with the wild strain, a dramatic increase in enzyme activity was observed for the mutant strain, although the increase in biomass was limited. DNA and amino acid sequence alignment revealed 11 nucleotide substitutions and 10 amino acid substitutions between the sldAB of strains L-6 and I-2-239. Simulation of the 3-D structure and prediction of active site residues and PQQ binding site residues suggested that these mutations were mainly related to PQQ binding, which was speculated to be favorable for the catalyzing capacity of glycerol dehydrogenase. RT-qPCR assay indicated that the transcription levels of sldA and sldB in the mutant strain were respectively 4.8-fold and 5.4-fold higher than that in the wild strain, suggesting another possible reason for the increased DHA productivity of the mutant strain.

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References

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