The utilization of

acid-tolerant bacteria
on ethanol
production from
kitchen garbage

INTRODUCTION
In order to achieve ethanol production from
kitchen garbage under non-sterilized
fermentation, the acidtolerant Zymomonas
mobilis named GZNS1 was selected and
applied in the fermentation system.
The utilization of acid-tolerant bacteria
combing with the utilization of distillerywaste
associated with the process can increase
ethanol production, save energy and reduce
the cost of ethanol production.

Materials and methods

Z. mobilis 10225 was obtained from China Center
of Industrial Culture Collection.
It was cultured on seed media with the
composition (g/L): glucose 100, yeast extract 10,
KH2PO4 1, (NH4)2SO4 1, MgSO4 0.5. The
microorganismwas maintained at 4 C and renewed
every 2 months
The base components of culture medium were 100
g kitchen garbage with 50 mL water, sterilized at
121 C for 15 min. The characteristics of kitchen
garbage shown in Table 1

Characteristic of the kitchen

garbage used in the experiment.

TS/% DS/
%

pH

Total Star Prot

suga ch/% ein/
r/%
%

Fat/
%

Cell
ulos
e/%

SS/%

17.2
2

5.62

62.6
8

18.0
6

2.26

14.6
4

2.58

46.1
2

15.5
6

Cultural conditions
Z. mobilis was firstly cultured for 24 h in the
culture medium under anaerobic conditions in
flasks. With a 10% (V/W) inoculum size, Z.
mobilis was added into the culture media
together with a dosage of glucoamylase (100
U/g wet mass) and protease (100 U/g wet
mass). Ethanol fermentation was conducted
within 150 Ml medium in 250 mL Erlenmeyer
flasks.
Anaerobic cultures were maintained at 30 C for
48 h in a stationary incubator, ethanol and
reducing sugar were measured at certain
intervals.

Ethanol fermentation under non-sterilized and

sterilized
conditions

In non-sterilized process, HCl was utilized

to adjust the pH of media to 4, then the
media were used to produce without
sterilization based on the procedure .
test was carried out without pH adjusted
under sterilized fermentation.
As the sterilized condition is concerned,
all implements such as flasks and media
were sterilized at 121 C for 15 min.

Ethanol fermentation with distillery

waste
The distillery waste from ethanol
production from kitchen garbage was
utilized to adjust the pH and water
content of the fermentation system.

Results
Strain isolation
Low pH of 4 was controlled in the non-sterilized condition to
prevent the potential contamination, thus the predominance of the
acid-tolerant bacteria was of great importance in such process.
Although Z. mobilis could perform fermentation in a wide range of

pH (37), to accomplish such processes successfully, the

acidtolerant bacteria with high growth rate is needed.
To obtain this goal, a special mutant was selected to meet the
special requirement.
Z. mobilis incubated in the seed medium was cultured in the
selection medium with pH of 4.
Then the large colonies were chosen for candidate for ethanol
fermentation under acid condition

Ethanol production under nonsterilized condition

The utilization of distillery waste for

ethanol production

 The reducing sugar under sterilized

process was a little higher at initial
than the nonsterilized one, that
might because other organism inside
the substrate would help to fasten
the hydrolysis of the substrate.
When the energy efficiency was
concerned, the addition of distillery
waste under non-sterilized condition
should be the alternative in the
following experiment (Fig. 4).