Methods for increasing the production of ethanol from microbial fermentation

Abstract

A stable continuous method for producing ethanol from the anaerobic bacterial fermentation of a gaseous substrate containing at least one reducing gas involves culturing in a fermentation bioreactor anaerobic, acetogenic bacteria in a liquid nutrient medium; supplying the gaseous substrate to the bioreactor; and manipulating the bacteria in the bioreactor by reducing the redox potential, or increasing the NAD(P)H TO NAD(P) ratio, in the fermentation broth after the bacteria achieves a steady state and stable cell concentration in the bioreactor. The free acetic acid concentration in the bioreactor is maintained at less than 5 g / L free acid. This method allows ethanol to be produced in the fermentation broth in the bioreactor at a productivity of greater than 10 g / L per day. Both ethanol and acetate are produced in a ratio of ethanol to acetate ranging from 1:1 to 20:1.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. patent application Ser. No. 10 / 311,655, filed Mar. 11, 2003, which is a US national phase of International Patent Application No. PCT / US01 / 23149, filed Jul. 23, 2001, which claims the benefit of the priority of U.S. Provisional Patent Application No. 60 / 220,794, filed Jul. 25, 2000, now abandoned.[0002]This invention was made with government support under Grant Nos. DE-FC04-94AL98770, 85X-TA046V, 85X-SX613V awarded by the US Department of Energy, and under Grant No. 96-MRC-1-0077 awarded by the US Department of Agriculture. The government has certain rights in the invention.BACKGROUND OF THE INVENTION[0003]The present invention is directed to improvements in microbial fermentation methods for the production of ethanol from a gaseous substrate containing at least one reducing gas using anaerobic (or facultative) acetogenic bacteria.[0004]Methods for producing ethanol, among other organic acids, al...

AI Technical Summary

Benefits of technology

[0009]In response to the need in the art, the present invention provides novel methods which are continuous, steady state methods and which result in ethanol concentrations greater than 10 g / L and acetate concentrations lower than about 8-10 g / L, while continuing to permit culture growth and good culture stability.

[0010]In one aspect, the invention provides a stable continuous method for producing ethanol from the anaerobic bacterial fermentation of a gaseous substrate. The method comprising the steps of culturing in a fermentation bioreactor anaerobic, acetogenic bacteria in a liquid nutrient medium and supplying to the bioreactor the gaseous substrate comprising at least one reducing gas selected from the group consisting of CO and H2. The bacteria in the bioreactor are manipulated by reducing the redox potential, or increasing the NAD(P)H TO NAD(P) ratio, in the fermentation broth after the bacteria achieves a steady state, e.g., a stable cell concentration, in the bioreactor. The free acetic acid concentration in the bioreactor is maintained at less than 5 g / L free acid. The culturing and manipulating steps cause the bacteria in the bioreactor to produce ethanol in a fermentation broth at a productivity greater than 10 g / L per day. Both ethanol and acetate are produced in the fermentation broth in a ratio of ethanol to acetate ranging from 1:1 to 20:1.

[0016]In another embodiment, the method of the invention includes preventing acclimation of said bacteria in said bioreactor to said amount of cobalt by maintaining a constant cobalt concentration and adjusting one or more parameters, such as gas rate, liquid rate, agitation rate and H2 gas partial pressure.

[0019]In yet another embodiment of the method, a step is provided in which inhibition by molecular acetic acid is reduced by increasing the aqueous feed rate when the molecular acetic acid present in the fermentation broth approaches or exceeds 2 g / L.

[0020]In another embodiment of the method, the manipulating step may include agitating the medium, bacteria and gaseous substrate in the bioreactor at a selected agitation rate. For example, reduction in the agitation rate reduces the amount of CO transferred to the fermentation broth. This reduction in the rate of CO transfer causes an increase in H2 conversion, so that the reducing gas, H2, is present in the bioreactor in excess of the growth requirements of the bacteria. The gas rate may also be similarly reduced to decrease the amount of CO transferred, thereby increasing H2 conversion, so that the reducing gas, H2, is present in the fermentation bioreactor in excess of the growth requirements of the bacteria.

Problems solved by technology

While this ability to produce ethanol is of interest, because of low ethanol productivity the “wild” bacteria cannot be used to economically produce ethanol on a commercial basis with minor nutrient manipulation the above-mentioned C. ljungdahlii strains have been used to produce ethanol and acetyl with a product ratio of 1:1 (equal parts ethanol and acetyl), but the ethanol concentration is less than 10 g / L, a level that results in low productivity, below 10 g / L·day.

In addition culture stability is an issue, primarily due to the relatively high (8-10 g / L) concentration of acetyl (2.5-3 g / L molecular acetic acid) in combination with the presence of ethanol.

As a result, the culture becomes unstable and fails to uptake gas and produce additional product.

Further, early work by the inventors showed difficulty in producing more than a 2:1 ratio of ethanol to acetyl in a steady state operation.

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