A Proteogenomic and Biochemical Classification of Novel Thermocellulolytic Bacillus licheniformis and Bacillus altitudinis isolates with Bio-Industry Potential Isolated from Yellowstone National Park
The industrial fermentation industry relies heavily upon unique microorganisms to carry out production of relevant bioproducts. In the case of second generation biofuels, there is a need for novel cellulolytic thermophiles. They have the potential to survive many environmental stressors while successfully generating high value bio-end-products. Yellowstone National Park is home to thousands of natural hot springs, each capable of sustaining microbiomes where these thermophiles of interest might be found. In a recent field study under permit# YELL-2015-SCI-6074, several of these hydrothermal areas were sampled using a non-invasive approach to isolate unknown culturable bacteria and fungi. From this, a total of 5 phenotypically unique cellulolytic colonies were identified as potential candidates. These 5 strains were subjected to biochemical assays, growth curve studies, temperature assays, and high throughput genomic sequencing. The 5 unknowns were classified as: Bacillus licheniformis YNP1-TSU, Bacillus licheniformis YNP2-TSU, Bacillus licheniformis YNP3-TSU, Bacillus altitudinis YNP4-TSU and Bacillus licheniformis YNP5-TSU. Thermophilic conditions were preferred by all isolates and peak growth was obtained from a range of 45°C to 60°C. Bio-reactor conditions were established under a defined cellulosic media to optimize all downstream testing. Cellulase activity from each strain was calculated and Bacillus licheniformis YNP5-TSU was found to be the most active at 0.1044 IU/ml. A proteomic study of Bacillus licheniformis YNP5-TSU also revealed that optimal cellulase production occurred only when a mature biofilm was able to develop and was dependent on the presence of magnesium. Through these classification methods all 5 cellulolytic thermophiles have proven to be potential candidates for further testing in fermentation analysis; providing significant evidence that novel cellulolytic thermophiles remain overlooked inside Yellowstone National Park.^
Joshua A OHair,
"A Proteogenomic and Biochemical Classification of Novel Thermocellulolytic Bacillus licheniformis and Bacillus altitudinis isolates with Bio-Industry Potential Isolated from Yellowstone National Park"
ETD Collection for Tennessee State University.