Essential Genes Encoding Conserved Metabolic Pathway Function in Autotrophic Solventogenic Clostridial Species

Abstract

Essential genes coding for the metabolic pathway of solventogenic autotrophic Clostridia were sequenced, and functionality was confirmed. The present invention utilizes a comparative inter-species approach to develop the minimum set of essential genes for metabolic function and estimate productivity in species of suspected solventogenic capability.

What is claimed is: 
     
         1 . An isolated and purified polynucleotide encoding a polypeptide sequence with a P value of 1×10 −20  or smaller when compared to SEQ ID NO. 1. 
     
     
         2 . An isolated and purified polynucleotide encoding a polypeptide sequence with a P value of 1×10 −20  or smaller when compared to the sequence of SEQ ID NO. 2. 
     
     
         3 . An isolated and purified polynucleotide encoding a polypeptide sequence with a P value of 1×10 −20  or smaller when compared to the sequence of SEQ ID NO. 3. 
     
     
         4 . A method of producing ethanol comprising: isolating and purifying anaerobic, ethanologenic microorganisms carrying the polynucleotide of  claim 1 ; fermenting syngas with said microorganisms in a fermentation bioreactor. 
     
     
         5 . A method of producing ethanol comprising: isolating and purifying anaerobic, ethanologenic microorganisms carrying the polynucleotide of  claim 2 ; fermenting syngas with said microorganisms in a fermentation bioreactor. 
     
     
         6 . A method of producing ethanol comprising: isolating and purifying anaerobic, ethanologenic microorganisms carrying the polynucleotide of  claim 3 ; fermenting syngas with said microorganisms in a fermentation bioreactor. 
     
     
         7 . A method of confirming high titer autotrophic solventogenesis function of a potentially commercially viable microorganism, said method comprising: sequencing the genome of said potentially commercially viable microorganism; comparing a resulting sequence of the genome of the microorganism to SEQ ID NO. 1. 
     
     
         8 . A method of prescreening natural isolates with suspected autotrophic solventogenesis function for high ethanol titer potential, said method comprising: isolating and enriching a sample containing said natural isolates; subjecting said sample to a polymerase chain reaction using at least one set of degenerate primers capable of hybridizing to one or more of the genes of  claim 1 ; separating the amplified product of the polymerase chain reaction based on size; and determining the presence of said genes based on the results of said separation.