A Proteogenomic and Biochemical Classification of Novel Thermocellulolytic Bacillus licheniformis and Bacillus altitudinis isolates with Bio-Industry Potential Isolated from Yellowstone National Park

Joshua A OHair, Tennessee State University

Abstract

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.^

Subject Area

Bioengineering|Microbiology|Bioinformatics

Recommended Citation

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" (2018). ETD Collection for Tennessee State University. Paper AAI10786967.
https://digitalscholarship.tnstate.edu/dissertations/AAI10786967

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