Skip to Content
Merck
  • Enhanced production of L-sorbose in an industrial Gluconobacter oxydans strain by identification of a strong promoter based on proteomics analysis.

Enhanced production of L-sorbose in an industrial Gluconobacter oxydans strain by identification of a strong promoter based on proteomics analysis.

Journal of industrial microbiology & biotechnology (2015-05-09)
Yudong Hu, Hui Wan, Jianghua Li, Jingwen Zhou
ABSTRACT

Gluconobacter oxydans is capable of rapidly incomplete oxidation of many sugars and alcohols, which means the strain has great potential for industrial purposes. Strong promoters are one of the essential factors that can improve strain performance by overexpression of specific genes. In this study, a pipeline for screening strong promoters by proteomics analysis was established. Based on the procedure, a new strong promoter designated as P B932_2000 was identified in G. oxydans WSH-003. The promoter region was characterized based on known genome sequence information using BPROM. The strength of P B932_2000 was further assessed by analysis of enhanced green fluorescent protein (egfp) expression and comparison with egfp expression by two commonly used strong promoters, P E. coli_tufB and P G. oxydans_tufB . Both quantitative real-time PCR and fluorescence intensities for egfp gene expression showed that P B932_2000 promoter is stronger than the other two. Overexpression of D-sorbitol dehydrogenase (sldh) by P B932_2000 in G. oxydans WSH-003 enhanced the titer and productivity of L-sorbose synthesis from D-sorbitol by 12.0 % and 33.3 %, respectively. These results showed that proteomics analysis is an efficient way to identify strong promoters. The isolated promoter P B932_2000 could further facilitate the metabolic engineering of G. oxydans.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Urea solution, BioUltra, ~8 M in H2O
Sigma-Aldrich
D-Sorbitol, BioUltra, ≥99.0% (HPLC)
Sigma-Aldrich
D-Sorbitol, FCC, FG
Sigma-Aldrich
D-Sorbitol, 99% (GC)
Sigma-Aldrich
Urea-12C, 99.9 atom % 12C
Sigma-Aldrich
Sorbitol F solution, 70 wt. % in H2O, Contains mainly D-sorbitol with lesser amounts of other hydrogenated oligosaccharides
Sigma-Aldrich
D-Sorbitol, ≥98% (GC)
Sigma-Aldrich
D-Sorbitol, ≥98% (GC), BioReagent, suitable for cell culture, suitable for plant cell culture
Sigma-Aldrich
D-Sorbitol, ≥98% (GC), BioXtra
Sigma-Aldrich
D-Sorbitol, ≥98% (GC), for molecular biology
Sigma-Aldrich
Urea solution, 40 % (w/v) in H2O
Sigma-Aldrich
Thiourea, ACS reagent, ≥99.0%
Sigma-Aldrich
Thiourea, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Bromophenol Blue, ACS reagent
Sigma-Aldrich
L-(−)-Sorbose, for biotechnological purposes, ≥98.0% (sum of enantiomers, HPLC)
Sigma-Aldrich
Bromophenol Blue, titration: suitable
Sigma-Aldrich
Urea, BioXtra, pH 7.5-9.5 (20 °C, 5 M in H2O)
Sigma-Aldrich
Urea, meets USP testing specifications
Sigma-Aldrich
Urea, BioUltra, for molecular biology, 99% (T)
Sigma-Aldrich
Urea, powder, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Urea, suitable for electrophoresis
Sigma-Aldrich
Urea, puriss., meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%, 99.0-101.0% (calc. on dry substance)
Sigma-Aldrich
Urea, ACS reagent, 99.0-100.5%
Supelco
Urea, 8 M (after reconstitution with 16 mL high purity water)
Sigma-Aldrich
Urea, ReagentPlus®, ≥99.5%, pellets
Sigma-Aldrich
Urea, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99%