Stabilized chlorine dioxide for contamination control in;fermentation

• US 8,846,357 B2
• Filed: 12/20/2012
• Issued: 09/30/2014
• Est. Priority Date: 12/20/2012
• Status: Expired due to Fees

• Current Assignee: Sustainable Technology Corporation
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Leana, Maria C, Lefebvre, Brian G
• Primary Examiner(s): Kosson, Rosanne
• Application Number:
  US13/721,103
• Publication Number:
  US 20140178956A1
• Time in Patent Office: 649 Days
• Field of Search: 435/161, 426/9, 127/37
• US Class Current: 435/161
• CPC Class Codes: A61L 2/18 Liquid substances or solutions comprising solids or dissolved gases
• CPC Class Codes: C08H 8/00 Macromolecular compounds derived from lignocellulosic materials pretreatment thereof B27N
• CPC Class Codes: C08L 97/02 Lignocellulosic material, e.g. wood, straw or bagasse manufacture of articles made from lignocellulosic material by dry processes B27N
• CPC Class Codes: C12H 1/00 Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages simulating ageing by flavouring C12G3/06
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 1/22 Processes using, or culture media containing, cellulose or hydrolysates thereof
• CPC Class Codes: C12N 1/36 Adaptation or attenuation of cells
• CPC Class Codes: C12N 2500/05 Inorganic components
• CPC Class Codes: C12N 9/2437 Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
• CPC Class Codes: C12P 7/065 with microorganisms other than yeasts
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

Though chlorine dioxide is generally used to control bacterial contamination, a method was developed which allows the use of stabilized chlorine dioxide (SCD) for controlling contamination during fermentation that uses the bacteria Zymomonas as the biocatalyst, even though Zymomonas is sensitive to chlorine dioxide. Parameters were identified for inoculating a composition for fermentation with Zymomonas cells after a time period has elapsed following SCD addition.

Claims

1. A fermentation method for controlling bacterial contamination in a fermentation process for making ethanol comprising a Zymomonas biocatalyst comprising:

a) providing a first fermentation medium lacking a cellulosic biomass hydrolysate selected from the group consisting of;

i) a defined medium, ii) a medium containing a non-cellulosic biomaterial sugar source, and iii) a clarified cellulosic biomass hydrolysate;

b) inoculating the first fermentation medium of a) with Zymomonas ethanologen cells to form a seed culture;

c) growing the Zymomonas cells in the seed culture of b);

d) providing a second fermentation medium or a saccharification reaction slurry;

e) inoculating the second fermentation medium or the saccharification reaction slurry of d) with the seed culture of c) to form a fermentation broth; and

f) growing the Zymomonas ethanologen cells in the fermentation broth under conditions whereby ethanol is produced;

wherein;

the second fermentation medium contains cellulosic biomass hydrolysate; and

wherein;

stabilized chlorine dioxide is added to at least one of i) the first fermentation medium of step a), ii) the saccharification reaction slurry of step d), or iii) the second fermentation medium of step d), and

wherein;

if the stabilized chlorine dioxide is added to the first fermentation medium of step a) the first fermentation medium is maintained at a temperature of at least 33°

C. for at least 8 hours prior to inoculation with the Zymomonas ethanologen; and

wherein;

if the stabilized chlorine dioxide is added to the saccharification reaction slurry of step d) the saccharification reaction slurry is maintained at a temperature of at least 33°

C. for at least 6 hours prior to inoculation with the Zymomonas ethanologen; and

wherein;

if the stabilized chlorine dioxide is added to the second fermentation medium of step d) the fermentation medium is maintained at a temperature of at least 33°

C. for at least 6 hours prior to inoculation with the Zymomonas ethanologen; and

wherein;

any one of the seed culture, the inoculated second fermentation medium or the fermentation broth comprising Zymomonas cells contains less than 5 g/L of lactic acid during the period of the growth of the Zymomonas cells.

2. The method of claim 1 wherein the seed culture, the inoculated fermentation medium and the fermentation broth are maintained at a temperature of less than 40°

C.

3. The method of claim 1 wherein the saccharification reaction slurry comprises at least one cellulase.

4. The method of claim 1 wherein the source of bacterial contamination is lactic acid producing bacteria.

5. The method of claim 1 wherein the cellulosic biomass is selected from the group consisting of corn cobs, corn husks, corn stover, grasses, wheat, wheat straw, barley straw, hay, rice straw, switchgrass, waste paper, sugar cane bagasse, sorghum, and woody plant cellulosic components.

6. The method of claim 1 wherein the concentration of stabilized chlorine dioxide in the seed medium of step a), the saccharification reaction slurry of step d), or the fermentation medium of step d) is initially at least 10 mg/kg.

7. The method of claim 1 wherein the saccharification reaction slurry of step d) or the fermentation medium of step d) comprising the stabilized chlorine dioxide contains at least 20% solids based on the dry weight of biomass to total mixture weight.