Biocatalysts for conversion of thermochemical waste streams
View Patent ↗Disclosed herein are microorganisms that have enhanced tolerance to toxic compounds found in thermochemical waste streams. Methods of utilizing carbon found in waste streams are also disclosed. Also presented herein are methods for detoxifying waste streams and methods of bioconversion of toxic waste stream materials into useful products.
1. A non-naturally occurring Pseudomonas cell that is capable of metabolizing at least 82% of the available carbon within 72 hours in a waste stream resulting from the pyrolysis of biomass wherein the Pseudomonas cell overexpresses active chaperone enzymes selected from the group consisting of GroES, GroEL and ClpB wherein the gene encoding for ClpB is operably linked to a tac promoter and has a nucleotide sequence that is greater than 85% identity to SEQ ID NO: 1 and encodes for an active ClpB enzyme having an amino acid sequence that is greater than 90% identity to SEQ ID NO: 2; and wherein the gene encoding for GroES is operably linked to a tac promoter and has a nucleotide sequence that is greater than 85% identity to SEQ ID NO: 3 and encodes for an active GroES enzyme having an amino acid sequence that is greater than 90% identity to SEQ ID NO: 4; and wherein the gene encoding for GroEL is operably linked to a tac promoter and has a nucleotide sequence that is greater than 85% identity to SEQ ID NO: 5 and encodes for an active GroEL enzyme having an amino acid sequence that is greater than 90% identity to SEQ ID NO: 6.
2. The cell of claim 1 wherein the genes are incorporated into the genome of the Pseudomonas cell.
3. The cell of claim 1 capable of an 83 fold or greater survival rate in comparison to the naturally occurring Pseudomonas from which it is derived after 12 hours of growth in a waste stream from the pyrolysis of biomass.
4. The cell of claim 1 able to grow in waste stream solutions containing concentrations of compounds that do not allow for the growth of the naturally occurring Pseudomonas from which it is derived from; the concentrations of compounds selected from the group consisting of greater than 7.5 times the concentration of aldehydes, 1.5 times the concentration of ketones, 3.5 times the concentration of acids, 3.5 times the concentration of phenolics, and 1.5 times the concentration of alcohols.
5. A method for metabolizing waste stream products from the pyrolysis of biomass comprising treating the waste stream products with the Pseudomonas cell of claim 1 .