J Ind Microbiol Biotechnol (2015) 42:543–551

DOI 10.1007/s10295-014-1572-7

BIOENERGY/BIOFUELS/BIOCHEMICALS

Bioethanol production from the dry powder of Jerusalem

artichoke tubers by recombinant Saccharomyces cerevisiae
in simultaneous saccharification and fermentation
Yi‑Zhou Wang · Shan‑Mei Zou · Mei‑Lin He ·
Chang‑Hai Wang 

Received: 30 September 2014 / Accepted: 18 December 2014 / Published online: 21 January 2015
© Society for Industrial Microbiology and Biotechnology 2015

Abstract  It has been found that recombinant Saccharo- fuels [16], people are eager to look for new energy to add
myces cerevisiae 6525 can produce high concentration of or replace the normal energy. In recent years, bioethanol
ethanol in one-step fermentation from the extract of Jeru- has attracted an increasing number of attentions as one of
salem artichoke tubers or inulin. However, the utilization the most important renewable and environmentally friendly
rate of raw materials was low and the fermentation process petroleum-based liquid fuels [5]. At present, derivatives
was costly and complicated. Therefore, in this study, after from food crops such as corn grain and sugar cane are gen-
the optimum processing conditions for ethanol production erally utilized in bioethanol production. However, it may
in fed-batch fermentation were determined in flask, the lead to huge competition between food provision and their
recombinant S. cerevisiae 6525 was first used to produce use for the production of bioethanol due to the limited sup-
ethanol from the dry powder of Jerusalem artichoke tubers ply of these crops, and some environmental problems such
in 5-L agitating fermentor. After 72 h of fermentation, as serious destruction of vital soil resources and food short-
around 84.3 g/L ethanol was produced in the fermentation ages may also be caused [9, 20]. In this case, an increasing
liquids, and the conversion efficiency of inulin-type sug- number of people around the world show great interest to
ars to ethanol was 0.453, or 88.6 % of the theoretical value look for new and cheap carbohydrate sources for bioetha-
of 0.511. This study showed high feasibility of bioethanol nol production [17].
industrial production from the Jerusalem artichoke tubers Jerusalem artichoke, as a cheap and inexpensive carbo-
and provided a basis for it in the future. hydrate alternative feedstock, has been studied for bioeth-
anol production since the 1950s. The tuber of Jerusalem
Keywords  Bioethanol · Saccharomyces cerevisiae · SSF · artichoke is rich in inulin which is present even more than
Jerusalem artichoke · Fed-batch fermentation 50 % in the dried tuber [19]. As a reserve carbohydrate,
inulin is a linear polymer of d-fructose containing a termi-
nal d-glucose which can be hydrolyzed to fructose, sucrose,
Introduction glucose and polymers of fructo-oligosaccharides by inuli-
nase [4]. Moreover, not only Jerusalem artichoke can resist
With the less and less energy on the earth and the pollu- many plant pests and diseases, but also grow well in barren
tion of the environment, especially the global warming and land with its growing traits of drought tolerance and saline
energy crisis problems caused by the combustion of fossil tolerance [2]. With these advantages, Jerusalem artichoke
can be used as a cheap and inexpensive carbohydrate alter-
Y.-Z. Wang and S.-M. Zou contributed equally to this work and
native feedstock for bioethanol production in addition to
should be considered co-first authors. its usual uses as single-cell oil or oligofructose syrup [10,
23, 28]. Remize et al. [22] used the extract from Jerusalem
Y.‑Z. Wang · S.‑M. Zou · M.‑L. He · C.‑H. Wang (*)  artichoke tuber as the substrate to produce ethanol by Sac-
College of Resources and Environmental Sciences,
charomyces cerevisiae strain. Ethanol was also produced
Nanjing Agricultural University, Nanjing 210095,
People’s Republic of China from Jerusalem artichoke tubers using recycled immo-
e-mail: chwang@njau.edu.cn bilized cells of Kluyveromyces fragilis [14]. Therefore,

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544 J Ind Microbiol Biotechnol (2015) 42:543–551

Jerusalem artichoke is a potential biomass source for etha- heated at 70 °C until reaching a constant weight in an oven
nol fermentation. and were milled to powder. The powder was stored at ambi-
Currently, the most commonly used strain in bioethanol ent temperature. The total sugar comprised about 62 % of
fermentation is S. cerevisiae which can produce high con- dry powder according to the phenol–sulfuric acid method
centration of ethanol from reducing sugar like glucose or [6].
fructose and has high ethanol tolerance [1, 11]. However,
ordinary S. cerevisiae cannot use the inulin directly to pro- Yeast strains
duce bioethanol. It needs pretreatment of hydrolyzing the
inulin included in Jerusalem artichoke tubers to fructose The recombinant S. cerevisiae 6525 with the exo-inuli-
and glucose by inulinase or acids before the process of nase gene from Penicillium janthinellum was already
ethanol fermentation [7], called SHF (separate hydrolysis constructed in our laboratory [25, 26]. It is chose as the
and fermentation), which is long-running and complicated. producer of ethanol from the dry powder of Jerusalem arti-
To produce bioethanol effectively and economically with choke tubers in this study. The recombinant S. cerevisiae
S. cerevisiae from Jerusalem artichoke, a recombinant S. 6525 was found to be able to produce 1.1 U mL−1 inulinase
cerevisiae strain which can produce exo-inulinase was con- activity within 72 h and 14.0 % (v/v) ethanol in fermenta-
structed successfully in our previous studies [21, 26]. Using tion medium containing inulin and 1 % (w/v) (NH4)2SO4 in
the recombinant S. cerevisiae, Jerusalem artichoke tuber a 5-L fermentor [25].
can be utilized to produce bioethanol in SSF (simultane-
ous saccharification and fermentation) with exo-inulinase Media
which can hydrolyze inulin to fructose effectively [3, 24].
Ethanol fermentation using the recombinant S. cerevisiae The strains were kept at 4 °C in medium YPD containing
from the raw extract of Jerusalem artichoke tubers contain- (in g/L) yeast extract, 10; peptone, dextrose, 20; and pep-
ing 26 % total sugars has also been carried out in our pre- tone, 20. The YPI medium which was used for exo-inuli-
vious studies [8]. The final ethanol concentration reached nase production by the recombinant S. cerevisiae 6525
102.1 g/L under the optimum conditions. However, using contained (in g/L): yeast extract, 10; peptone, 20; and inu-
pure inulin or raw extract of Jerusalem artichoke tubers as lin, 20. The medium used for ethanol fermentation by the
substrate in previous studies is too costly and complicated, recombinant S. cerevisiae 6525 contained 240 g/L dry pow-
and has low feasibility of bioethanol industrial production. der of Jerusalem artichoke tubers, the fermentation medium
In our present study, to develop cost-effective and sus- composition was varied based on the fermentation experi-
tainable technologies for industrial bioethanol production ments. The pH value of the fermentation medium was
and increase the utilization rate of Jerusalem artichoke between 5.3 and 5.6. For flask study, pH was adjusted to
tubers, the dry powder of Jerusalem artichoke tubers was 5.5.
used directly to produce bioethanol by the recombinant S.
cerevisiae. In addition, the fermentation medium contain- Preparation of the exo‑inulinase produced by the
ing the dry powder of Jerusalem artichoke tubers usually recombinant S. cerevisiae 6525 carrying the inulinase gene
has a high viscosity, which is detrimental to high-gravity
ethanol fermentation. Therefore, the optimum cooking pre- 200  μl of recombinant S. cerevisiae 6525 inoculum was
treatment of medium before fermentation and the optimum added to 100 mL YPI medium, cultured at 30 °C and
fermentation conditions in flask based on fed-batch fermen- 180 rpm for 72 h to produce enough exo-inulinase. The cell
tation were investigated in this study. Then, scale-up test in density was 1 × l08 cells/mL after counting the cells using
5-L agitating fermentor was carried out based on the opti- a hemocytometer. The cell culture was used as both the
mum conditions of flask fermentation, which could provide exo-inulinase and ethanol producer.
a basis for our next pilot fermentation test in 50- and 500-L
agitating fermentor for industrial production. Cooking pretreatment of fermentation medium

Fermentation medium was cooked at the optimum tem-

Materials and methods perature before fermentation to decrease the medium vis-
cosity and accelerate the release of inulin in the dry pow-
Materials der of Jerusalem artichoke tubers. 100 mL of fermentation
medium containing 240 g/L dry powder of Jerusalem arti-
Jerusalem artichoke was harvested in maturity from Dafeng choke tubers in the 250-mL flask was heated at 30, 50, 70,
in China at the end of November. The tubers were washed 80, 90 °C in water rocker (150 r/min) for 30 min and heated
in running water and cut into pieces. Then, they were at 115 °C for 20 s in high-pressure steam sterilization,

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J Ind Microbiol Biotechnol (2015) 42:543–551 545

Table 1  Four kinds of fed-batch methods and the concentration of 5, 10, 15 and 20 % (v/v) of seed culture were inocu-
the dry powder of Jerusalem artichoke tubers in initial time, 24, 48 lated into the fermentation medium in a 250-mL flask with
and 72 h
100 mL of fermentation medium containing 240 g/L dry
Fed-batch method Initial time 24 h 48 h 72 h powder of Jerusalem artichoke tubers and 10 g/L yeast
(g/L) (g/L) (g/L) (g/L) extract, respectively. The flask was cultured at 30 °C and
① Fed-batched at 24 h 240  180 rpm, for 96 h at anaerobic conditions. As fermentation
② Fed-batched at 24 240
300 300 300
280 300 300 proceeded, we fed-batched in two equal amounts at 24 and

③ Fed-batched at 24 240
and 48 h 48 h to make the final concentration of dry powder reach
270 300 300 300 g/L.
and 48 h The initial medium pH was adjusted at a unit of 0.5–4.0,
④ Fed-batched at 24, 240 270 300 330 4.5, 5.0, 5.5 and 6.0, respectively. 10 % (v/v) seed culture
48 and 72 h
was inoculated into a 250-mL flask with 100 mL fermenta-
tion medium containing 240 g/L dry powder of Jerusalem
artichoke tubers and 10 g/L yeast extract. The flask was
cultured at 30 °C and 180 rpm, for 96 h at anaerobic con-
respectively. After cooling the medium to room tempera- ditions. As fermentation proceeded, we fed-batched in two
ture, 10 % (v/v) seed culture was inoculated into the 250- equal amounts at 24 and 48 h to make the final concentra-
mL flask with the final volume of 100 mL fermentation tion of dry powder reach 300 g/L.
medium. The flask was incubated at 30 °C and 180 rpm, for 10 % (v/v) seed culture was inoculated into a 250-mL
96 h at anaerobic conditions. flask of fermentation medium (pH 4.5) with the bottle load
of 75, 100, 125 and 150 mL (the final fermentation medium
Fed‑batch fermentation for ethanol production volume was 30, 40, 50 and 60 %), respectively. The flask
was incubated at 30 °C and 180 rpm, for 96 h at anaerobic
10 % (v/v) seed culture was inoculated into a 250-mL flask conditions. As fermentation proceeded, we fed-batched in
with 100 mL fermentation medium containing 240 g/L dry two equal amounts at 24 and 48 h to make the final concen-
powder of Jerusalem artichoke tubers. The flask was incu- tration of dry powder reach 300 g/L.
bated at 30 °C and 180 rpm, for 96 h at anaerobic condi- 10 % (v/v) seed culture was inoculated into a 250-mL
tions. As the fermentation proceeded, additional Jerusalem flask with 125 mL fermentation medium (pH 4.5) contain-
artichoke dry powder was supplemented. Four kinds of fed- ing 240 g/L dry powder of Jerusalem artichoke tubers and
batch methods are shown in Table 1: fed-batched at 24 h, 10 g/L yeast extract. The flask was incubated at 28, 30, 32,
the final concentration reached 300 g/L; fed-batched at 34 and 36 °C, respectively, 180 rpm, for 96 h at anaerobic
24 h, the concentration reached 280 g/L, then fed-batched conditions. As fermentation proceeded, we fed-batched in
at 48 h again, the final concentration reached 300 g/L; fed- two equal amounts at 24 and 48 h to make the final concen-
batched in two equal amounts at 24 and 48 h, the final con- tration of dry powder reach 300 g/L.
centration reached 300 g/L; and fed-batched in three equal
amounts at 24, 48 and 72 h, the final concentration reached Scale‑up test in 5‑L agitating fermentor
330 g/L. The final ethanol concentration and residual total
sugar in the fermented media were determined as described After fermentation condition was optimized in flask,
below. fermentation curves were investigated using 5-L agitat-
ing fermentor (i.d. 15.1 cm, height 28.0 cm, 3.0 L work-
Ethanol production from the dry powder of Jerusalem ing volume, BIOTECH-5JG, Zhenjiang, China) under the
artichoke tubers in 250‑mL flask optimum conditions. The initial medium (2.5 L) contain-
ing 240 g/L dry powder of Jerusalem artichoke tubers and
10 g/L yeast extract, 10 g/L peptone, 10 g/L corn steep liq- 10 g/L yeast extract was used with the initial pH adjusted
uor, 10 g/L NH4NO3, and 10 g/L NH4H2PO4 were added to 4.5. The fermentation medium was cooked at 115 °C
to the fermentation medium, respectively, and no nitrogen for 20 s before being used. 10 % (v/v) seed culture was
source was added to the control group in the experiment. inoculated into the fermentation medium, and the culture
10 % (v/v) seed culture was inoculated into a 250-mL flask was carried out at 34 °C and 180 rpm, for 96 h at anaerobic
containing 100 mL fermentation medium. The flask was conditions. As fermentation proceeded, we fed-batched in
cultured at 30 °C and 180 rpm, for 96 h at anaerobic con- two equal amounts at 24 and 48 h so as to make the final
ditions. As fermentation proceeded, we fed-batched in two concentration of dry powder reach 300 g/L. The final etha-
equal amounts at 24 and 48 h to make the final concentra- nol concentration and residual total sugar in the fermented
tion of dry powder reach 300 g/L. liquids were determined as described below.

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546 J Ind Microbiol Biotechnol (2015) 42:543–551

Table 2  Analysis of the dry powder of Jerusalem artichoke tubers

Dried weight (%) Total sugar (%) Reducing sugar (%)

The dry powder of Jerusalem artichoke tuber 100 61.9 ± 2.4 5.9 ± 1.2

Table 3  Effect of different Cooking method Viscosity of medium (mPa s) Total sugar after cooking (g/L) Ethanol (g/L)
cooking pretreatments on
ethanol fermentation from 30 °C 30 min 1.78 × 105 125.17 ± 2.28 53.34 ± 4.04
the dry powder of Jerusalem
50 °C 30 min 1.24 × 105 131.12 ± 2.63 56.00 ± 3.61
artichoke tubers
70 °C 30 min 1.55 × 104 134.05 ± 2.95 61.34 ± 3.51
80 °C 30 min 1.33 × 104 138.16 ± 2.09 63.00 ± 2.65
90 °C 30 min 1.29 × 104 139.02 ± 2.11 62.34 ± 1.53
115 °C 20 s 1.32 × 104 144.08 ± 2.05 66.34 ± 3.06

Analytical methods high (5.4 ton/ha) [13]. The tuber of Jerusalem artichoke is

rich in inulin which can be used as substrate for ethanol pro-
Ethanol concentration was determined using a biosensor duction. The previous studies have indicated that the extract
(SBA-40C) from Biology Institute of Shandong Academy of Jerusalem artichoke tubers or inulin can be fermented into
of Science (Jinan, China). Viscosity was determined using high concentration of ethanol by the recombinant S. cerevi-
a rotational viscosimeter (NDJ-1) from Shanghai Jinke siae 6525 [8, 25]. However, the utilization rate of raw mate-
Company (Shanghai, China). The total sugar was measured rials was low and the fermentation process was costly and
according to the phenol–sulfuric acid method [6]. Reducing complicated. To increase the utilization rate of Jerusalem
sugar, mainly fructose in the fermentation liquids detected artichoke tubers, this study used the dry powder of Jerusa-
in our previous researches [25, 26], was quantified by the lem artichoke tuber directly to produce ethanol. After analy-
3,5-dinitrosalicylic acid method [15]. sis of the dry powder of Jerusalem artichoke tubers used in
Statistical analysis of ethanol concentration, total sugar this study, it can be observed from the results in Table 2 that
and reducing sugar was processed by SPSS 20.0. The com- the dry powder was containing 61.9 ± 2.4 % (w/w) of total
parisons were tested after analysis of variance (one-way sugar and 5.9 ± 1.2 % (w/w) of reducing sugar.
ANOVA) using Duncan’s test. Data are presented as the Since the fermentation liquor containing the dry powder
mean  ± S.E. for each treatment. Different letters indicate of Jerusalem artichoke tubers has a high viscosity, cooking
statistical difference at p < 0.05. pretreatment of fermentation medium was carried out in
The ethanol yield in this study was evaluated by the con- this study before the process of fermentation. The results in
version efficiency of inulin-type sugars to ethanol: Table 3 showed that the viscosity of medium in treatments
heated at 70, 80, 90 °C for 30 min and 115 °C for 20 s was
Yp = P/(C × 62 %)
much lower than the other treatments. Although the viscos-
where P was the total ethanol produced (g/L), C was the ity of medium did not continue to decrease at the treatment
final concentration of the dry powder of Jerusalem artichoke cooked at 115 °C for 20 s, the sugar concentration dis-
tubers in medium (g/L), 62 % was the total sugar in the dry solved in the medium was higher than the other treatments.
powder of Jerusalem artichoke tubers determined experi- As can be seen in Fig. 1, after 48 h of fermentation, the fer-
mentally by the method described in the “Materials and mentation medium heated at 70, 80 and 90 °C, which could
methods” section. And the ethanol yield in the figure was provide germicidal effect similar to pasteurization, had rel-
expressed as the percentage of the theoretical value of 0.511. atively higher ethanol yield than the treatments heated at 30
and 50 °C, but no significant difference of the ethanol yield
was observed between them. And the highest ethanol yield
Results and discussion of 87.2 % was achieved at the treatment cooked at 115 °C
for 20 s. It was possible that the medium cooked at 115 °C
The most suitable cooking pretreatment of fermentation for 20 s could improve the lixiviating effect and kill more
medium for ethanol production other harmful microorganisms. In this way, the fermenta-
tion efficiency was improved under the sterile conditions.
Jerusalem artichoke is a non-food crop which grows exten- Thus, in this study, the fermentation medium was cooked at
sively in many parts of China and the yield of its tuber is very 115 °C for 20 s before fermentation.

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J Ind Microbiol Biotechnol (2015) 42:543–551 547

Fig. 1  Effect of different cook-

ing pretreatments on ethanol
fermentation from the dry
powder of Jerusalem artichoke
tubers. Ethanol yield was
expressed as the percentage of
the theoretical value of 0.511

The most suitable fed‑batch method for ethanol 160

Total sugar of method 1
fermentation 140
Ethanol of method 1
Total sugar of method 2
Ethanol of method 2
120 Total sugar of method 3
Ethanol, Total sugar(g/L)

Zhang et al. [27] used the Saccharomyces sp. W0 to produce Ethanol of method 3
Total sugar of method 4
ethanol from the hydrolysate of inulin, and fed-batched inu- 100 Ethanol of method 4
lin at 48 and 96 h. Under this condition, 14.6 mL ethanol
80
in 100 mL fermented medium was produced within 120 h
of fermentation. In our study, as fermentation proceeded, 60

we fed-batched the dry powder of Jerusalem artichoke

40
tubers to keep a suitable concentration of total sugar in the
medium. The results in Fig. 2 showed that the method 3 20

with fed-batched in two equal amounts at 24 and 48 h was 0

the most suitable condition for ethanol production by the 0 12 24 36 48 60 72 84 96

recombinant S. cerevisiae 6525. Under the condition, etha- Culture time(h)

nol concentration increased continuously within 72 h of the
fermentation and then decreased. The highest ethanol con- Fig. 2  Time course of fed-batch fermentation for ethanol production
centration of 79.0 g/L was achieved at 72 h and 90.8 % of from the dry powder of Jerusalem artichoke tubers at different fed-
batch methods
total sugar was used. As can be seen in Table 4, the ethanol
yield of method 3 was 83.12 %, which was the highest one
than the other three methods. In accordance with our obser- some metabolites had a protective effect on ethanol pro-
vations, supplement could dissolve in fermentation liquor duction. The method with more than two times fed-batched
quickly and the treatments with fed-batched in two equal would be left behind for further research.
amounts at 24 and 48 h could keep a relatively low viscos-
ity. This demonstrates that the dry powder of Jerusalem arti- Effect of different nitrogen sources on ethanol production
choke tubers can also be fermented into high concentration
of ethanol by the recombinant S. cerevisiae 6525 in fed- After different nitrogen sources were added to the fer-
batch fermentation. The treatment with fed-batched in three mentation medium, the recombinant S. cerevisiae 6525
equal amounts at 24, 48 and 72 h achieved the highest final was inoculated into the fermentation medium in the 250-
dry powder concentration of 330 g/L and ethanol concen- mL flask. It can be observed from the results in Fig. 3a
tration of 83.67 g/L at 84 h, but the ethanol yield was only that when 10 g/L yeast extract was added to the fermen-
79.87 %, which was lower than the treatment with method tation medium, the highest ethanol yield of 83.12 % was
3. It may be possible that in the fermentation anaphase, achieved, which was higher than the other treatments. In

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548 J Ind Microbiol Biotechnol (2015) 42:543–551

Table 4  Effect of four kinds of Fed-batch method Theoretical content of total sugar (g/L) Ethanol (g/L) Ethanol yield (%)
fed-batch methods on ethanol
fermentation from the dry Method 1 186  71.34  75.06 
powder of Jerusalem artichoke
Method 2 186  75.67  79.61 
tubers
Method 3 186  79.00  83.12 
Method 4 205  83.67  79.87 

Fig. 3  Effect of nitrogen
source, inoculation, initial
medium pH, medium volume
and temperature on ethanol
production in fed-batch fermen-
tation from the dry powder of
Jerusalem artichoke. Ethanol
yield was expressed as the
percentage of the theoretical
value of 0.511. a Nitrogen
source, b inoculation, c initial
medium pH, d medium volume,
e temperature

addition, NH4H2PO4, as inorganic nitrogen, was much from 10 to 20 % after 72 h of fermentation (data not
cheaper than other organic nitrogen for ethanol industrial shown), and the ethanol yield of the treatment with 5 %
production, which had a good prospect of utilization and inoculum was lower than the other treatments. Therefore,
will be studied in further research. Finally, for flask study, the inoculation number we chosen in this study was 10 %.
we chose yeast extract as nitrogen source in the experiment.
Effect of initial medium pH on ethanol production
Effect of inoculation on ethanol production
The results in Fig. 3c indicated that the ethanol concen-
It can be seen from the data in Fig. 3b that there was no sig- tration in the medium was the highest while the initial
nificant difference for the ethanol yield at the inoculation medium pH value was 4.5. Under the conditions, 84.33 g/L

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J Ind Microbiol Biotechnol (2015) 42:543–551 549

ethanol was achieved after 72 h of fermentation, and the

ethanol yield was over 88.73 %. Much lower ethanol yield
was obtained at the pH values of 4.0, and no significant dif-
ference for the ethanol yield was found at the pH values
of 5.0, 5.5 and 6.0. This means that the initial medium pH
value of 4.5 was the most suitable pH for ethanol produc-
tion by the recombinant S. cerevisiae 6525 and was also the
optimal pH of the recombinant exo-inulinase, which is con-
sistent with the results in Wang et al. [26]. However, the
reason for this phenomenon still needs to be further studied
from the impact of the medium pH on the growth of recom-
binant S. cerevisiae 6525, different processes of respiration
and exo-inulinase production, especially the formation of
some by-products in the process of fermentation.
Fig. 4  Time course of fed-batch fermentation for ethanol production
Effect of medium volume on ethanol production from the dry powder of Jerusalem artichoke tubers with recombinant
S. cerevisiae 6525 in 5-L agitating fermentor
As can be seen in Fig. 3d, treatments with medium vol-
ume of 30, 40, 50, 60 % produced 79.0 ± 2.4, 83.7 ± 1.5,
86.3 ± 1.5 and 84.3 ± 3.1 g/L ethanol after 72 h of fermen- tubers was used for ethanol production by the recombinant
tation, respectively. The highest ethanol yield of 90.83 % S. cerevisiae 6525 in 5-L agitating fermentor as described
was achieved at the treatment with medium volume of in “Materials and methods”. The results in Fig. 4 showed
50 %. It indicated that excess oxygen supply was detrimen- that the process of ethanol production proceeded very
tal to ethanol production by the recombinant S. cerevisiae rapidly in the first 36 h and the ethanol concentration still
6525. Thus, the most suitable medium volume we tempo- increased slowly from 36 to 60 h with fed-batched in two
rarily chose would be 50 % in this study. equal amounts at 24 and 48 h. The highest ethanol concen-
tration of 84.3 g/L was obtained at 72 h, and the conver-
Effect of temperature on ethanol production sion efficiency of inulin-type sugars to ethanol was 0.453,
or 88.6 % of the theoretical value of 0.511 [12]. The total
The flask was incubated at 28, 30, 32, 34 and 36 °C, sugar in the fermentation liquids was decreased continu-
respectively. It was found that when the cultured tempera- ously after the start of the fermentation, while the reduc-
ture was 34 °C, the ethanol concentration in the fermenta- ing sugar in the fermentation liquids was increased slightly
tion medium reached the highest. It can also be seen from within 12 h of fermentation and then still kept a relatively
the result in Fig. 3e that the most suitable cultured tempera- lower concentration until 60 h of fermentation, which
ture for ethanol production by the recombinant S. cerevi- helped prevent substrate inhibition. After that, the reduc-
siae 6525 was 34 °C, at which the highest ethanol yield of ing sugar was decreased until the end of fermentation. At
93.98 % was obtained. The reason for this phenomenon the end of the fermentation, only 14.6 g/L total sugar and
may be related to the inulinase activity, the growth of 5.5 g/L reducing sugar were remained in the fermentation
recombinant S. cerevisiae 6525 and the solubility of the dry liquids.
powder of Jerusalem artichoke tubers, which needs further At present, as the dry powder of Jerusalem artichoke
studies. tubers is rich in inulin and has a much lower processing
cost for industrial production, more and more researchers
Scale‑up test in 5‑L agitating fermentor are interested in using the dry powder of Jerusalem arti-
choke tubers as substrate for ethanol production. Zhang
In the previous studies [18], the flour of Jerusalem arti- et al. [27] used the tuber meal of Jerusalem artichoke
choke tubers was used to produce ethanol by A. niger 817 [the concentration of the tuber meal in the fermentation
and S. cerevisiae 1200 in simultaneous saccharification and medium was 50 % (w/v)] to produce ethanol by Saccha-
fermentation, and 20.1 % of ethanol was obtained within romyces sp. W0 in 5-L agitating fermentor, 12.05 mL etha-
120 h of fermentation after complementing flour at 15 and nol in 100 mL fermented medium was obtained within
24 h. However, the process of fermentation was compli- 144 h, and the ethanol productivity was 0.319 ± 0.9 g of
cated and time consuming, and the utilization rate of sugar ethanol/g of sugar. However, because the fermentation
was only 80 %. In our study, after fermentation condition medium containing the dry powder of Jerusalem artichoke
was optimized in flask, dry powder of Jerusalem artichoke tubers usually has a high viscosity, the utilization rate of

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550 J Ind Microbiol Biotechnol (2015) 42:543–551

the sugar included in dry power is too low and the process 2. Bajpai PK, Bajpai P (1991) Cultivation and utilization of Jerusa-
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syrup production. Enzyme Microb Tech 13:359–362
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In our study, the recombinant S. cerevisiae 6525 was first A, Cervelli C, Ruffoni B (2011) Amplified fragment length poly-
used to produce ethanol from the dry powder of Jerusalem morphism markers for DNA fingerprinting in the genus Salvia.
artichoke tubers. The result indicated that proper cooking Plant Biosyst 145:274–277
4. Byun SM, NAHM BH (1978) Production of fructose from Jerusa-
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ter performance for high-gravity ethanol fermentation. In formation under very high gravity fermentation condition. Bio-
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Acknowledgments We gratefully acknowledge the support 14. Margaritis A, Bajpai P (1981) Repeated batch production of etha-
from the Hi-Tech Research and Development Program of China nol from Jerusalem artichoke tubers using recycled immobilized
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Conflict of interest  The authors declare that they have no conflict 16. Mohan SV, Babu VL, Sarma PN (2008) Effect of various pre-
of interest. treatment methods on anaerobic mixed microflora to enhance
biohydrogen production utilizing dairy wastewater as substrate.
Ethical approval  This article does not contain any studies with Bioresour Technol 99:59–67
human participants or animals performed by any of the authors. 17. Mohanty SK, Behera S, Swain MR, Ray RC (2009) Bioethanol
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