Beruflich Dokumente
Kultur Dokumente
Some substances in the solutions can have inhibitory effect on the fermentation by microorganisms since the
used cultivation media are complex. Their composition is not completely defined as it varies due to factors like
techniques used, climate, type of employed fertilizers, water availability etc.
Dilution- Molasses must be diluted to below 25° brix as yeast start to ferment quickly at this concentration
Sedimentation- is performed to prevent any incrustation in the pipelines or distillation towers due to ash
content in molasses greater than 10%. Special chelating agents can also be employed to remove the solids
causing incrustation
Addition of org & inorganic compounds- This is done to offset the negative effect of salts which in turn
increase the osmotic pressure. Yeast strains resistant to salts are also developed
Microfiltration- To remove the impurities that stick to the surface of the biocatalyst when immobilized cells
are used
• Nitrogen source:
Urea is the most suitable. Gaseous
ammonium increases the pH of the
medium & ammonium sulfate can
lead to incrustation
• Phosphorous source:
Diammonium phosphate is used
pH : 4-4.5
Temperature: 30°C
MICROORGANISMS INVOLVED
Saccharomyces cerevisiae
Convert hexose into pyruvate by Glycolysis which is finally
reduced to ethanol generating 2 moles of ATPs under
anaerobic conditions
Can tolerate high concentrations of ethanol up to 150g/L
Ethanol production is coupled with yeast cell growth, which
means yeast must be produced as a co-product
By products like glycerol, organic acids are also produced
Zymomonas mobilis
Anaerobe, gram-negative bacteria which produces ethanol via ED
pathway converting 1 mol of hexose into 2 mol of ethanol, but releasing
only 1 mol of ATP
Lower cell yield due to lower energy yield of bacterium, increasing the
amount of ethanol from the substrate (97%)
High ethanol tolerance (100g/L) & higher optimum production temp.
Drawbacks
Highly specific substrate spectrum : glucose, fructose & sucrose
decreasing ethanol yield
It’s biomass is not acceptable to be used as animal feed, which
generates a problem for biomass disposal
Continuous ethanol fermentation is oscillatory which can increase the
average residual sugar but decrease the ethanol yield
MICROORGANISMS INVOLVED
• During the first 12-24 h, yeast cells multiply rapidly aerobically by consuming oxygen present in the
mash
• Decrease in alcohol formation along with insignificant yeast growth at the final stage ( 48-72h)
BATCH FERMENTATION- MELLE BOINOT PROCESS
Molasses or Yeast
Propagation Yeast reutilization
sugarcane juice
Fermenter
wort
Adjustment of Wine
Weighing & Decantation &
pH using H2SO4 Fermentation
sterilization centrifugation
& brix to 14-22
Yeast reuse results in a decrease in new growth with more sugar available for ethanol production & an increase in the
yield from 2 to 7%.
Traditional yields – 1-3 g/L
High concentration of yeasts (44g) & high supplementation of yeast extract (28g/L)at low ethanol content (60g/L)if
employed in glucose based medium, ethanol productivity is as high as 21g/L
Operating procedures for fermentation include
The stillage represents one of the distillation product • Washing and disinfection of the fermenter
streams during the subsequent ethanol recovery step that • Filling up of the fermenter with the culture medium and
contains a significant amount of water and a much reduced sterilization of such medium
amount of ethanol. The addition of stillage to the culture • Inoculation of microbial cells
broth can lead to lower water consumption & reduction of • Fermentation
stillage volume to be treated. • Unloading of bioreactor content at the end of the
cultivation process.
CONTINUOUS FERMENTATION
Strain : S.cerevisiae
Sterilization: 100°C, 30 min
Viscosity : 1000-2000 cps
METABOLIC ENGINEERING
S. cerevisiae cannot utilize lactose directly, whereas the yeast Kluyveromyces lactis can
utilize lactose but cannot perform an efficient alcohol fermentation
To develop an efficient lactose fermenting yeast, the β-galactosidase gene from K. lactis,
along with the cloned lactose permease gene, was introduced into S. cerevisiae, leading
to the fermentation of lactose.