Essential genes encoding conserved metabolic pathway function in autotrophic solventogenic clostridial species

• US 20120003652A1
• Filed: 07/01/2010
• Published: 01/05/2012
• Est. Priority Date: 07/01/2010
• Status: Active Grant

• Current Assignee: Synata Bio, Inc.
• Original Assignee: Coskata, Inc.
• Inventors: Reeves, Andrew, Yin, Fenglin
• Granted Patent: US 8,987,431 B2
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12P 7/06 Ethanol, i.e. non-beverage
• CPC Class Codes: C12P 7/065 with microorganisms other than yeasts
• CPC Class Codes: C12Y 102/02004 Carbon-monoxide dehydrogenase (cytochrome b-561)(1.2.2.4)
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

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.

Claims

1. An isolated and purified polynucleotide encoding a polypeptide sequence with a P value of 1×

10<sup>−

20</sup> or smaller when compared to SEQ ID NO. 1.

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.

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.

2. An isolated and purified polynucleotide encoding a polypeptide sequence with a P value of 1×

10<sup>−

20</sup> or smaller when compared to the sequence of SEQ ID NO. 2.

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.

3. An isolated and purified polynucleotide encoding a polypeptide sequence with a P value of 1×

10<sup>−

20</sup> or smaller when compared to the sequence of SEQ ID NO. 3.

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.