# US 2014/0154727 A1 · Geobacter Strains That Use Alternate Organic Compounds, Methods of Making, and Methods of Use Thereof

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- Number: US 2014/0154727 A1
- Document ID: US20140154727A1
- Status: Published application
- Kind: A1
- Application number: 14/070,748
- Filed: 2013-11-04
- Published: 2014-06-05
- Earliest claimed: 2008-08-08
- Assignees: University of Massachusetts
- Inventors: Derek R. Lovley; Zarath Morgan Summers; Shelley Annette Haveman; Mounir Izallalen
- Web page: https://patentgrep.com/patent/US20140154727A1

## CPC

- C12N 1/20: Bacteria; Culture media therefor
- C12N 1/36: Adaptation or attenuation of cells
- H01M 8/16: Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
- H01M 4/90: Selection of catalytic material
- H01M 8/18: Regenerative fuel cells, e.g. redox flow batteries or secondary fuel cells
- Y02E 60/50: Fuel cells
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- H01M: PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

In preferred embodiments, the present invention provides new isolated strains of a Geobacter species that are capable of using a carbon source that is selected from C3 to C12 organic compounds selected from pyruvate or metabolic precursors of pyruvate as an electron donor in metabolism and in subsequent energy production. The wild type strain of the microorganisms has been shown to be unable to use these C3 to C12 organic compounds as electron donors. The inventive strains of microorganisms are useful for improving bioremediation applications, including in situ bioremediation (including uranium bioremediation and halogenated solvent bioremediation), microbial fuel cells, power generation from small and large-scale waste facilities (e.g., biomass waste from dairy, agriculture, food processing, brewery, or vintner industries, etc.) using microbial fuel cells, and other applications of microbial fuel cells, including, but not limited to, improved electrical power supplies for environmental sensors, electronic devices, and electric vehicles.

## Claims

### Claim at position 1

1-14. (canceled)

### Claim 15

A method of making an isolated adapted strain of an electricigenic microbe that can use an alternative organic compound as an electron donor wherein the alternative organic compound is not preferably used as an electron donor by a parent strain of the electricigenic microbe, comprising the steps of:
a. providing a first vessel that contains a first culture, the first culture comprising the parent strain of the electricigenic microbe, and
a first growth medium that includes
a preferred organic compound as an electron donor and
a corresponding electron acceptor;
b. inoculating a second vessel that contains
a second growth medium that includes
an alternative organic compound that replaces the preferred organic compound as an electron donor and
a corresponding electron acceptor
with an aliquot of the parent strain of the electricigenic microbe to form an adapted culture;
c. monitoring a growth rate of the adapted culture, noting the start of an exponential phase, and continuing until the growth rate is in mid or late exponential phase;
d. setting n=1, where n is an integer from 1 to 100, inclusive;
e. incrementing n by 1 according to the relation n=n+1;
f. transferring an aliquot of the adapted culture to another vessel that contains the second growth medium to inoculate an nth adapted culture;
g. monitoring the growth rate of the nth adapted culture, noting the start of an exponential phase, and continuing until the growth rate is in mid or late exponential phase;
h. repeating steps e-g unless
either the time period from inoculation until the start of the exponential phase of the nth adapted culture is more than 0.4 times the time period from inoculation until the start of the exponential phase of the (n−1)th adapted culture; or
the increase in the growth rate of the nth adapted culture compared to its predecessor is less than that of the (n−1)th adapted culture compared to its predecessor, or
the doubling time of the nth adapted culture in the second growth medium is greater than or equal to the doubling time of the parent strain in the first growth medium; and
l. isolating the adapted strain of the electricigenic microbe from the nth adapted culture.

### Claim 16 (depends on 15)

The method of claim 15 wherein the preferred organic compound is acetate and the alternative organic compound is pyruvate or a C3 to C12 metabolic precursor of pyruvate.

### Claim 17 (depends on 15)

The method of claim 15 wherein the parent strain is Geobacter sulfurreducens strain identified as ATCC deposit ATCC-51573 and the isolated adapted strain is Geobacter sulfurreducens strain identified as ATCC deposit PTA-10250.

### Claim 18 (depends on 17)

The method of claim 17 wherein the preferred organic compound is acetate and the alternative organic compound is lactate.

### Claim 19 (depends on 15)

The method of claim 15 wherein the parent strain is the 1-23 strain of Geobacter sulfurreducens, the preferred organic compound is lactate and the alternative organic compound is glucose.

### Claim 20 (depends on 19)

The method of claim 19 wherein the adapted strain of Geobacter sulfurreducens is the strain identified as ATCC deposit PTA-10250.

### Claim 21 (depends on 15)

The method of claim 15 wherein the parent strain is the 1-23 strain of Geobacter sulfurreducens, the preferred organic compound is lactate and the alternative organic compound is glycerol.

### Claim 22 (depends on 21)

The method of claim 21 wherein the adapted strain of Geobacter sulfurreducens is the strain identified as strain ZMS-353.

### Claim 23 (depends on 15)

The method of claim 15 wherein the parent strain is the 1-23 strain of Geobacter sulfurreducens, the preferred organic compound is lactate and the alternative organic compound is xylose.

### Claim 24 (depends on 23)

The method of claim 23 wherein the adapted strain of Geobacter sulfurreducens is the strain identified as strain ZMS-453.