title: High-throughput screening and characterization of xylose-utilizing, ethanol-tolerant thermophilic bacteria for bioethanol production.
abstract: OBJECTIVE
  To develop a high-throughput assay for screening xylose-utilizing and ethanol-tolerant thermophilic bacteria owing to their abilities to be the promising ethanologens.
  
  
  RESULTS
  Based on alcohol oxidase and peroxidase-coupled enzymatic reaction, an assay was developed by the formation of the coloured quinonimine to monitor the oxidation of ethanol in the reaction and calculate the concentration of ethanol. This assay was performed in 96-well microtitre plate in a high-throughput and had a well-linear detection range of ethanol from 0 up to 2·5 g l(-1) with high accuracy. The assay was then verified by screening soil samples from hot spring for xylose-utilizing and ethanol production at 60°C. Three isolates LM14-1, LM14-5 and LM18-4 with 3-5% (v/v) ethanol tolerance and around 0·29-0·38 g g(-1) ethanol yield from xylose were obtained. Phylogenetic and phenotypic analysis showed that the isolates clustered with members of the genus Bacillus or Geobacillus subgroup.
  
  
  CONCLUSIONS
  The developed double enzyme-coupled, high-throughput screening system is effective to screen and isolate xylose-utilizing, ethanol-producing thermophilic bacteria for bioethanol production at the elevated temperature.
  
  
  CONCLUSIONS
  Our research presented a novel high-throughput method to screen thermophilic bacteria for producing ethanol from xylose. This screening method is also very useful to screen all kinds of ethanologens either from natural habitats or from mutant libraries, to improve bioethanol production from lignocellulosic feedstocks.
authors: Key Laboratory of Food Nutrition and Safety, Tianjin University of Science & Technology, Tianjin, China.
authors: X
authors: X
authors: Qi
authors: Y
authors: Y
authors: Zhang
authors: R
authors: R
authors: Tu
authors: Y
authors: Y
authors: Lin
authors: X
authors: X
authors: Li
authors: Q
authors: Q
authors: Wang
mesh_terms: D000429:Alcohol Oxidoreductases / Q000032:analysis; D001419:Bacteria / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D056804:Biofuels / Q000382:microbiology*; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation; D057166:High-Throughput Screening Assays; D045482:Hot Springs / Q000382:microbiology; D006358:Hot Temperature; D007218:Industrial Microbiology; D009195:Peroxidase / Q000032:analysis; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012988:Soil Microbiology; D014994:Xylose / Q000378:metabolism*
chemical_list: D056804:Biofuels; D012336:RNA, Ribosomal, 16S; D000431:Ethanol; D014994:Xylose; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D009195:Peroxidase