CyTA - Journal of Food

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Effect of mixed fermentation (Jiuqu and

Saccharomyces cerevisiae EC1118) on the quality
improvement of kiwi wine

An-Jun Chen, Yun-Yun Fu, Cheng Jiang, Jiang-Lin Zhao, Xiao-Ping Liu, Lu Liu, Ji
Ma, Xing-Yan Liu, Guang-Hui Shen, Mei-Liang Li & Zhi-Qing Zhang

To cite this article: An-Jun Chen, Yun-Yun Fu, Cheng Jiang, Jiang-Lin Zhao, Xiao-Ping Liu, Lu
Liu, Ji Ma, Xing-Yan Liu, Guang-Hui Shen, Mei-Liang Li & Zhi-Qing Zhang (2019) Effect of mixed
fermentation (Jiuqu and Saccharomyces�cerevisiae EC1118) on the quality improvement of kiwi
wine, CyTA - Journal of Food, 17:1, 967-975, DOI: 10.1080/19476337.2019.1682678

To link to this article: https://doi.org/10.1080/19476337.2019.1682678

© 2019 The Author(s). Published with Published online: 11 Nov 2019.

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CYTA – JOURNAL OF FOOD
2019, VOL. 17, NO. 1, 967–975
https://doi.org/10.1080/19476337.2019.1682678

Effect of mixed fermentation (Jiuqu and Saccharomyces cerevisiae EC1118) on the

quality improvement of kiwi wine
An-Jun Chen a, Yun-Yun Fua, Cheng Jianga, Jiang-Lin Zhaoa, Xiao-Ping Liua, Lu Liua, Ji Mab, Xing-Yan Liua, Guang-
Hui Shena, Mei-Liang Lia and Zhi-Qing Zhanga
a
College of Food Science, Sichuan Agricultural University, Ya’an, China; bSichuan Nongxingyuan Agricultural Development Co. Ltd., Ya’an,
China

ABSTRACT ARTICLE HISTORY

The aim of this study was to investigate the effect of mixed fermentation of Jiuqu and Saccharomyces Received 5 August 2019
cerevisiae on the quality of kiwifruit wine. The results showed that total titratable acidity, methanol and Accepted 14 October 2019
organic acids of kiwi wine fermented by Jiuqu + Saccharomyces cerevisiae EC1118 were significantly KEYWORDS
lower (p < 0.05) than that in other groups. There was no significant difference in the content of flavor Kiwi wine; mixed
substances in kiwi wines fermented by a mixed group and Saccharomyces cerevisiae EC1118 alone, but fermentation; Jiuqu;
the sensory evaluation showed a significantly large difference. On the sensory scale, kiwi wine fermen- Saccharomyces cerevisiae;
ted by mixed fermentation had a unique presentation and typical bouquet, better than those fermented quality
by Jiuqu and Saccharomyces cerevisiae EC1118 alone. In conclusion, mixed fermentation of Jiuqu +
Saccharomyces cerevisiae EC1118 could improve the quality of the kiwi wine, which provides a promising PALABRAS CLAVE
Vino de kiwi; fermentación
application prospection for kiwi wine production. mixta; Jiuqu; Saccharomyces
cerevisiae; calidad

Efecto de la fermentación mixta (jiuqu y Saccharomyces cerevisiae EC1118) en la

mejora de la calidad del vino de kiwi
RESUMEN
El presente estudio se propuso investigar el efecto de la fermentación mixta con jiuqu
y Saccharomyces cerevisiae en la calidad del vino de kiwi. Los resultados mostraron que la acidez
valorable total, el metanol y los ácidos orgánicos del vino de kiwi fermentado con jiuqu +
Saccharomyces cerevisiae EC1118 fueron significativamente menores (p < 0.05) que en otros grupos.
No se detectaron diferencias significativas en el contenido de sustancias aromatizantes presentes en
los vinos de kiwi fermentados ya sea por el grupo mixto o por Saccharomyces cerevisiae EC1118 solo,
pero en la evaluación sensorial se constató una diferencia significativamente grande. En este
sentido, en la escala sensorial se constató que el vino de kiwi fermentado por fermentación mixta
tuvo una presentación única y un aroma típico, mejor que los vinos fermentados con jiuqu solo
o con Saccharomyces cerevisiae EC1118. En conclusión, la fermentación mixta del vino de kiwi con
jiuqu + Saccharomyces cerevisiae EC1118 podría mejorar la calidad, lo cual implica una prospección
de aplicación prometedora para su producción.

1. Introduction a popular kiwi-based product in Asia that increases its com-

Kiwifruit (Actinidia chinensis Planch), originates in China and
belongs to Actinidiaceae family (Luh & Wang, 1984), is
a popular fruit for its pleasant tasted and profuse nutrition,
being especially rich in vitamin C, organic acids, tannins, car-
bohydrates, pectin, essential amino acids, trace elements as
well as inositol and lutein (Nishiyama et al., 2004; Tavarini,
Degl’Innocenti, Remorini, Massai, & Guidi, 2008; Wei &
Guohua, 2015). There are a lot of health benefits associated
with kiwi such as antioxidant and anti-cancer activity, improv-
ing immunity, reducing blood pressure and blood fat, protect-
ing against diabetes and treatment of burns (Leontowicz et al.,
2016). In recent years, the production of kiwi fruit has been
remarkably increased due to the development of cultivation
techniques and production management all over the world,
which in turn lead to its overproduction (López-Vázquez et al.,
2012). There is an urgent demand to find a solution for utilizing
this overproduction for economic benefits. Kiwi wine is
mercial value.
Saccharomyces cerevisiae is one of the most commonly used
yeast for the fermentation of kiwi juice in China; however, there
are a few problems associated with S. cerevisiae fermented kiwi
wine such as lack of ample fruitiness, high acidity, low typical-
ity, high methanol content. Some researchers believed that
S. cerevisiae is not the ideal one for producing kiwi wine
(Kang et al., 2011). Mixed microorganism fermentation may
improve the quality of kiwi fruit wine as there are many suc-
cessful examples on other fruit: Englezos et al. (2016) reported
that compared to pure fermentation, mixed fermentation with
Starmerella bacillaris and S. cerevisiae improved the quality of
Barbera wine because of the synergistic effect of the two
microorganisms, the glycerine content can be increased to
8.8–9.8 g/L and acetic acid content can be reached up to 0.4
g/L (Gobbi et al., 2013). Garcia et al. (2002) found that mixed
fermentation with Debaryomyces vanriji and S. cerevisiae

CONTACT An-jun Chen chen_anjun@sicau.edu.cn College of Food Science, Sichuan Agricultural University, Xinkang road 46, Ya’an, China
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/tcyt.
© 2019 The Author(s). Published with license by Taylor & Francis Group, LLC.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
968 A.-J. CHEN ET AL.
significantly improved the concentration of several acids, alco-
hol, ester, and terpinol, and therefore better volatile profile of
grape wine was detected by mixed fermentation. Minnaar,
Jolly, Paulsen, Du Plessis, and Van Der Rijst (2017) reported
that the titratable acid of Kei-apple juice fermented by
Schizosaccharomyces pombe and S. cerevisiae yeasts decreased
by 70% compared to single culture fermentations. According
to these reported researches, kiwi wine produced with mixed
fermentation may improve its final quality.
Jiuqu is a kind of fermentation starter for the production of
Chinese sweet rice wine (CSRW), which is a traditional Chinese
alcohol beverage, possessing a desirable flavor and high nutri-
tional value, including peptides, oligosaccharides, vitamins,
amino acids, and organic acids (Jiang et al., 2016; Shen et al.,
2010; Shen, Ying, Li, Zheng, & Hu, 2011; Wei et al., 2017). The
starters are the mixtures of yeasts, molds, and bacteria (Bora,
Keot, Das, Sarma, & Barooah, 2016; Lv, Weng, Zhang, Rao, & Ni,
2012), which produce plenty of enzymes for cellular metabo-
lisms and subsequent small molecule generation, which con-
tribute to final quality of the products (Cai et al., 2018).
Therefore, the aim of this work was to investigate the
organic acid, flavor substance, methanol and sensory quality
of kiwi wines produced by the mixed fermentation of Jiuqu
and S. cerevisiae.
2. Materials and methods
2.1. Kiwi handling procedure and microorganisms
Fifteen kilograms kiwi fruit were purchased from the market
(Ya’an, Sichuan, China) at commercial maturity and with uniform
shape and size, the pH was 2.93 ± 0.06, total acidity was 14.27 ±
0.46 g/L citric acid, soluble solids content was 9.63 ± 0.55 °Brix
and organic acid content was 31.28 ± 2.03 mg/g FW (Fresh
Weight). The fruits were pulped with a domestic juicer.
Pectinase (60 mg/L) (40 u/mg, from Aspergillus niger, Beijing
Solarbio Science & Technology Co., Ltd., China) was added to
the pulp, and incubated at 40°C for 4.5 h, and then centrifuged at
4000 rpm for 10 min (Heraeus Multifuge X3R, Thermo Scientific,
USA), the supernatant was obtained with four layers of nylon,
followed by addition of SO2 (80 mg/L). Twenty percent (v/w) of
sucrose was added to kiwi juice and then stored at 4°C until use.
Jiuqu was purchased form Angel Yeast Co., Ltd (Yichang,
Hubei, China). S. cerevisiae EC1118 was obtained from
Xinmiao Winery (Ya’an, Sichuan, China).
2.2. Fermented kiwi juice and fermentation procedure
The Kiwi juice was divided into three groups as follows:
(1) Kiwi juice fermented by S. cerevisiae EC1118: 600 mL
of kiwi juice and 2 g dried S. cerevisiae EC1118 was
added to a sealed jar and fermented at room tem-
perature according to the manufacturer’s instructions.
(2) Kiwi wine fermented by Jiuqu: 600 mL of kiwi juice
and 6 g Jiuqu was added to a sealed jar and fermen-
ted at room temperature (22°C) according to the
manufacturer’s instructions.
(3) Kiwi wine fermented by Jiuqu + S. cerevisiae EC1118:
600 mL of kiwi juice and 6 g Jiuqu and 2 g dried
S. cerevisiae EC1118 was added to a sealed jar and
fermented at room temperature according to the
manufacturer’s instructions.
Transit operation for all kiwi wines was carried out after
fermenting for 8 days to remove dregs and then further
fermented at 22°C for 8 days. The raw wine was obtained
after removing dregs. All raw wines were filled with glass jars
and aged for 60 days at room temperature. All treatments
were conducted in triplicate. Kiwi wines were monitored by
recording the loss in mass.
2.3. Physico-chemical parameters
The analyses included the alcohol percentage, total titratable
acidity (TTA), total sugar (TS) and ascorbic acid content (AA)
of wine, according to the methods described by Chung, Son,
Park, Kim, and Lim (2008). Clarity and chromaticity were
measured by UV-1800PC visible spectrophotometer (UV-
1600PC, MAPADA, China) at 680 nm, 520 nm, respectively.
All treatments were conducted in triplicate.
Determination of condition and method validation of
HPLC-DAD and GC-FID of organic acids and methanol
content
Organic acids were quantified using a published HPLC-DAD
method (Barreca, Bellocco, Caristi, Leuzzi, & Gattuso, 2011;
Coelho et al., 2018; Mesquita & Monteiro, 2018), with slight
modification. The HPLC-DAD determination of organic acids
was performed using an Agilent HPLC model (Agilent
Technologies, PaloAlto, CA, USA). For the test method of
organic acids, an Agilent Eclipse plus C18 reserved-phase
column (5μm, 250 mm × 4.6 mm) and an Agilent Eclipse
plus C18 guard column (5μm, 20 mm × 4.6 mm) were used.
The sample injection volume was 10 μL, the temperature
was 30°C, the detection wavelength was 210 nm, the mobile
phase consisted of deionized water-dipotassium phosphate
(A: 100:0.02, v/v) and methanol (B), and the flow rate was
1 mL/min. Initial condition was A-B (98:2, v/v), linearly chan-
ged to A-B (95:5, v/v) at 8 min and then run for 10 min.
Standard solution of organic acids with different concen-
trations (0.0025–0.64 g/L) was prepared. The standard curves
of different organic acids were plotted with the concentration
of the standard solution as the abscissa and the peak area as
the ordinate. For the sample determination, the sample was
diluted 10 times, and passed through the 0.22 μm organic
filter membrane and then injected into HPLC-DAD.
Gas chromatograph (7890A/59750) (Agilent Technologies,
Palo Alto, CA, USA) was used for the determination of
methanol. Chromatographic separation was performed on
an Agilent HP-INNOWAX capillary column (0.25 mm ×
30 m). Helium was used as the carrier gas at a flow rate of
0.5 mL/min, the split ratio was 50:1, the flow rate of hydro-
gen was 40 mL/min, with a corresponding airflow rate of
40 mL/min, and makeup gas flow rate was 25 mL/min,
accompanied with hydrogen flame ionization detector.
Detector and injector temperatures were set at 220°C. The
following GC oven temperature program was applied: 40°C
for 4 min, 3.5°C/min to 96°C, 96°C hold for 2 min, 20°C/min
to 200°C, 200°C hold for 10 min; the sample injection volume
was 1 μL. Detection and quantitation limits, precision, recov-
ery and linearity were assessed in terms of analytical meth-
ods established in previous studies (Coelho et al., 2018;
Gupta, 2015; Mesquita & Monteiro, 2018).
Standard solution of methanol of different concentrations
(0.01–0.64 g/L) was prepared. The internal standard of
4-methyl-2-pentanol was added to the standard solution and
 CYTA - JOURNAL OF FOOD 969

the final concentration was made to 0.2 g/L. Methanol was
tested according to the method mentioned previously. The
standard curve of methanol was plotted with the concentration
of the standard solution as the abscissa and the peak area ratio
of methanol to 4-methyl-2-pentyl alcohol as the ordinate.
2.4. Validation of the applied method
In this study, the optimal chromatographic condition was
determined for the isolation of organic acids，methanol and
4-methyl-2-pentanol. As shown in Figure 1(a,b), the standard
samples of organic acids and methanol were isolated in the
optimal condition. The standards of different concentrations
of organic acids and methanol had a good linear relationship
with the peak area (R2 > 0.9997), which is in accordance with
the report by Coelho et al. (2018) that the R2 must over 0.99.
According to the results, the established method could be
used to assay organic acids and methanol.
The limits of detection (LOD) and quantification (LOQ) were
used to assay the ability of the established method to detect and
quantify compounds at low concentrations (Magnusson, 2014).
Table 1 shows that the LOD of organic acids was in the range of
0.0004–0.0032 g/L, and LOQ was in the range of 0.0016–0.0063
g/L. The LOD and LOQ of methanol were less than 0.01 g/L and
0.04 g/L, respectively. The results were in agreement with pre-
vious other reports (Chinnici, Spinabelli, Riponi, & Amati, 2005;
Coelho et al., 2018; Eyéghé-Bickong, Alexandersson, Gouws,
Young, & Vivier, 2012). The standard samples of organic acids
and methanol were added to measure their recovery rate, which
was in the range of 93.90%~103.2%, and all RSD of repeatability
was less than 2.77%. Therefore, the method could be used to
detect the organic acids and methanol in this study.
2.5. Analysis of aroma compounds
The wine aroma compounds were isolated and pre-
concentrated following a solid-phase extraction (SPE)

Figure 1. HPLC-DAD chromatogram of organic acid standards (a) and GC-FID chromatogram of methanol and 4-methyl-2-pentanol standards (b). Peak 1: Oxalic
acid, peak 2: Tartaric acid, peak 3: Malic acid, peak 4: Lactic acid, peak 5: Acetic acid, peak 6: Citric acid. Peak 7: Methanol, peak 8: 4-methyl-2-pentanol.
Figura 1. Cromatograma HPLC-DAD de normas de ácido orgánico (a) y cromatograma GC-FID de metanol y estándares de 4-metil-2-pentanol (b). Pico 1: ácido
oxálico; pico 2: ácido tartárico; pico 3: ácido málico; pico 4: ácido láctico; pico 5: ácido acético; pico 6: ácido cítrico; pico 7: metanol; pico 8: 4-metil-2-pentanol.
970 A.-J. CHEN ET AL.

Table 1. The retention time, regression equation, linear range, R2, LOD, LOQ, recovery and repeatability of organic acids and methanol determination.
Tabla 1. Tiempo de retención, ecuación de regresión, rango lineal, R2, LOD, LOQ, recuperación y repetibilidad de ácidos orgánicos y determinación de metanol.
Recovery Repeatability
Retention Range (100%) (% RSD)
Organic acids time (min) Calibration curve (N = 3) (g L−1) (N = 6) R2 LOD (g L−1) LOQ (g L−1) (N = 6) (N = 6)
Oxalic 2.075 Y = 7069.20X+ 8.2963 0.0025–0.64 0.9997 0.0004 0.0016 98.9 0.4
Tartaric 2.187 Y = 666.62X– 2.4895 0.0025–0.64 0.9998 0.0032 0.005 98.3 1.03
Malic 2.443 Y = 451.72X– 0.7297 0.0025–0.64 0.9998 0.0032 0.005 93.9 2.65
Lactic 2.819 Y = 186.63X+ 0.4499 0.0025–0.64 0.9999 0.005 0.0063 97.7 2.76
Acetic 3.089 Y = 316.36X+ 0.7903 0.0025–0.64 0.9997 0.0008 0.0025 93.7 2.12
Citric 3.448 Y = 659.30X+ 0.4944 0.0025–0.64 0.9999 0.0008 0.0032 95.1 0.64
Methanol FID
Methanol 6.388 Y = 1.645X+ 0.0138 0.01–0.64 1 0.01 0.04 103.2 1.94
DAD, photodiode array detector; R2, linear correlation coefficient; LOD, limit of detection limit; LOQ, limit of quantification.
DAD, detector de matriz de fotodiodos; R2, coeficiente de correlación lineal; LOD, límite de detección; LOQ, límite de cuantificación.

procedure (García-Carpintero, Sánchez-Palomo, Gallego, & applied to check the statistical significance of differences
González-Viñas, 2011) and then assayed by gas chromato- among different treatments with Statistics 22.0 (SPSS Inc.,
graphy-mass spectrometry (GC-MS) as previously described Chicago, IL, USA). Origin Lab original 8.5 pro software was
(Velázquez, Zamora, Álvarez, Álvarez, & Ramírez, 2016). used for plotting.

2.6. Sensory evaluation

Sensory evaluation of kiwi wine was followed by Friedman test
as described by Sherwood (1993) to compare the differences in
different kiwi wines. A panel of 15 experts who had extensive
experience in tasting kiwi wine were employed for the sensory
test. The kiwi wines were presented in clear flute-shaped wine
glasses. Synthetic ranking test of different kiwi wines was
carried out by experts and graded according to the following
criteria: 1-common, 2-good and 3-perfect. According to the
evaluation results, order and rank sum were attained.
2.7. Statistical analysis
The determinations were conducted in triplicates. One-way
analysis of variance (ANOVA) by Tukey test (p < 0.05) was
3. Results and discussion
3.1. Physico-chemical parameters
Figure 2 shows the fermentation curve of kiwi wine by
different treatments at 22°C. It can be seen that kiwi wine
produced with Jiuqu + S. cerevisiae EC1118 had the fastest
rate of fermentation than that produced by S. cerevisiae
EC1118 and Jiuqu alone. The release of CO2 reached to the
highest level on the third day in mixed fermentation group,
while in S. cerevisiae and Jiuqu fermented group, the peak
value was at fourth and fifth day, respectively. Moreover, the
peak CO2 value in the mixed group was much higher than
that in other groups.
Table 2 shows the physico-chemical parameters of the
produced kiwi wines. For ethanol content, Jiuqu fermented

Figure 2. Fermentation curve of kiwi juice fermented by different processes at 22°C (mean ± SD, n = 3). Kiwi juice fermented by Jiuqu ( ), EC1118 ( ) and
Jiuqu + EC1118 ( ).
Figura 2. Curva de fermentación del jugo de kiwi fermentado por diferentes procesos a 22°C (media ± DE, n = 3). Jugo de kiwi fermentado por Jiuqu ( ),
EC1118 ( ) y Jiuqu + EC1118 ( ).
 CYTA - JOURNAL OF FOOD 971

kiwi wine recorded the highest ethanol content, that was
13.47 ± 0.02%. The ethanol in Jiuqu + S. cerevisiae EC1118
group was 11.94 ± 0.02%, and it was 11.14 ± 0.02% in
S. cerevisiae EC1118 fermented group. But Jiuqu treated
group showed relatively higher residual sucrose content.
The high ethanol and residual sucrose content might be
caused by ethanol production ability of different microor-
ganisms (Hong et al., 2016), the results indicated that the
micro-flora in Jiuqu showed higher ethanol production abil-
ity than S. cerevisiae EC1118.
For TTA content, lower TTA content was observed in
mixed fermentation group than other groups, which may
indicate a low sour taste in the final products.
Clarity serves as one of the key factors in judging overall
wine quality (Valentin, Parr, Peyron, Grose, & Ballester, 2016).
For the clarity parameters, that were transmittance and
absorbency, mixed fermentation showed higher transmit-
tance (98.7 ± 0.21) but lower absorbency (0.029 ± 0.00),
suggesting higher clarity of kiwi wine produced by Jiuqu +
S. cerevisiae EC1118 fermentation.
3.2. Organic acids and methanol
The contents of organic acids from all wines were analyzed
by HPLC-DAD, and their values are presented in Table 3.
Total organic acids were in the range from 16336.69 to
22952.05 mg/L for all the wines. Organic acid significantly
affected the pH of kiwi wine, which contributed the rough
and harsh taste to the final products (Colangelo, Torchio, De
Faveri, & Lambri, 2018). The kiwi wine produced with mixed
fermentation showed significantly lower (p < 0.05) total
organic acids than the kiwi wines produced by pure
fermented groups, which were accord with the titratable
acid content (Ribéreau-Gayon, Glories, Maujean, &
Dubourdieu, 2006, Table 2).
Malic acid was the most abundant organic acids detected
in produced kiwi wines, which amounted to more than 55%
of total organic acids, followed by citric, tartaric and acetic
content. The content of malic and citric acids in kiwi wine
fermented by Jiuqu + S. cerevisiae EC1118 was lower, while
the lactic acid content was higher when compared with that
in kiwi wine fermented by S. cerevisiae EC1118 alone, which
may be caused by the convert of malic acid and citric acid to
lactic acid and CO2 (Suárez-Lepe, Palomero, Benito,
Calderón, & Morata, 2012). Besides, lactic acid has lower
acidity and higher stability than that of malic acid (Zhao,
Fan, Hang, & Yang, 2006), the high content of lactic acid in
mixed fermentation group increased the stability and sen-
sory profile of produced kiwi wine.
Acetic acid is an important microbial growth inhibitor, that
is produced during normal yeast metabolism in biotechnolo-
gical processes (Palma, Guerreiro, & Sá-Correia, 2018). The
content of acetic in Jiuqu group was the highest one, which
was about 4 times than that by mixed fermentation and about
10 times higher than that fermented by S. cerevisiae EC1118
alone. This can be attributed to the high metabolism of
Rhizopus in Jiuqu (Londoño-Hernández et al., 2017).
The methanol content in kiwi wines measured by GC-FID
was listed in Table 3. For all kiwi wines，the methanol con-
tent ranged from 162.64 to 212.05 mg/L, and the order of
methanol content in different fermentations was S. cerevisiae
EC1118 > Jiuqu > mixed fermentation (p < 0.05). The decline
of methanol content in mixed fermentation group was
decreased by 49.41mg/L in comparison with S. cerevisiae

Table 2. The effect of different processes on the physico-chemical parameters of kiwi wine.
Tabla 2. Efecto de diferentes procesamientos en los parámetros fisicoquímicos del vino de kiwi.
Sample (n = 3)
Parameters measured Jiuqu Yeast EC1118 Jiuqu + Yeast EC1118
Alcohol (% v/v) 13.47 ± 0.02a 11.14 ± 0.02c 11.94 ± 0.02b
TTA (g/L) 10.51 ± 0.01a 10.15 ± 0.01b 9.14 ± 0.03c
Content of Vc (g/L) 0.50 ± 0.01b 0.61 ± 0.01a 0.49 ± 0.03b
Residual sucrose (g/L) 6.21 ± 0.17a 6.06 ± 0.05ab 5.93 ± 0.07b
Transmittance (T%) 98.4 ± 0.06b 83.2 ± 0.1c 98.7 ± 0.21a
Absorbency (A) 0.040 ± 0.00b 0.169 ± 0.00a 0.029 ± 0.00c
Super indexes a, b, and c in the same row indicate significant differences in different treatments according to the least
significant difference test (p < 0.05). TTA, total titratable acid; Vc, Vitamin C.
Los superíndices a, b y c en la misma fila indican diferencias significativas en los diferentes tratamientos de acuerdo con la
prueba de diferencia menos significativa (p < 0.05). TTA, ácido valorable total; Vc, vitamina C.

Table 3. Organic acids and methanol content of kiwi wines.

Tabla 3. Ácidos orgánicos y contenido de metanol de los vinos de kiwi.
Treatment (n = 3)
Organic acids (DAD 210 nm) Jiuqu Yeast EC1118 Jiuqu + Yeast EC1118
Oxalic (mg/L) 250.11 (±1.76)a 166.59 (±3.99)c 195.59 (±0.49)b
Tartaric (mg/L) 1,839.55 (±2.41)b 2,305.74 (±1.03)a 1,709.38 (±1.89)c
Malic (mg/L) 10,009.46 (±4.32)b 13,196.84 (±3.59)a 9,023.11 (±2.64)c
Lactic (mg/L) 1,182.60 (±2.65)a 439.16 (±2.32)c 517.84 (±0.32)b
Acetic (mg/L) 4,674.50 (±0.77)a 474.52 (±0.52)c 1,267.42 (±2.71)b
Citric (mg/L) 3,936.36 (±2.06)b 6,369.20 (±5.49)a 3,623.32 (±1.32)c
Total organic acids 21,912.59 (±6.17)b 22,952.05 (±6.70)a 16,336.69 (±6.92)c
Methanol FID
Methanol (mg/L) 183.91 (±5.30)b 212.05 (±3.06)a 162.64 (±2.43)c
Super indexes a, b, and c in the same row indicate significant differences in different treatments according to the least significant difference test (p < 0.05).
Los superíndices a, b y c en la misma fila indican diferencias significativas en diferentes tratamientos de acuerdo con la prueba de diferencia menos significativa
(p < 0.05).
972 A.-J. CHEN ET AL.

EC1118 group. Methanol is a kind of toxin substance to
humans (Zhang, Lin, Chai, & Barnes, 2015), its formation
depends on many factors such as raw materials, pectinase,
fermentation temperature (Cabaroglu, 2005). Microbial
metabolism and the degradation of pectin by pectinase
could generate methanol (Amerine & Ough, 1980;
Cordonnier, 1987, Ribéreau-Gayon et al., 2006). At the same
time, Rogerson, Vale, Grande, and Silva (2000) pointed out
that S. cerevisiae can replace glycine to produce methanol in
raw materials. In this study, the methanol content of wine

Figure 3. Content of volatile flavor substances in different kiwi wines. (mean ± SD, n = 3). Kiwi juice fermented by Jiuqu ( ), EC1118 ( ) and Jiuqu + EC1118
( ).
Figura 3. Contenido de sustancias de sabor volátiles en diferentes vinos de kiwi (media ± DE, n = 3). Jugo de kiwi fermentado por Jiuqu ( ), EC1118 ( )
y Jiuqu + EC1118 ( ).

Figure 4. GC-MS chromatogram of different samples fermented by Jiuqu, EC1118 and Jiuqu + EC1118.
Figura 4. Cromatograma GC-MS de diferentes muestras fermentadas por Jiuqu, EC1118 y Jiuqu + EC1118.
 CYTA - JOURNAL OF FOOD 973

Table 4. The details of identified volatile components for each sample.

Tabla 4. Detalles de los componentes volátiles identificados para cada muestra.
Content (mg/L)
Volatile flavor Jiuqu + EC1118 Jiuqu EC1118
Ethyl esters
Ethyl caproate 1.83 ± 0.81 0.94 ± 0.25 1.88 ± 0.53
Ethyl 2-furoate 0.16 ± 0.01 0.24 ± 0.07 0.49 ± 0.07
Ethyl benzoate 0.43 ± 0.20 0.32 ± 0.07 0.33 ± 0.09
Diethyl succinate 2.65 ± 0.28 2.44 ± 0.40 3.56 ± 1.07
Ethyl caprylate 14.69 ± 2.92 5.08 ± 2.53 15.02 ± 1.39
Ethyl phenylacetate 0.18 ± 0.03 0.20 ± 0.01 0.17 ± 0.03
Phenethyl acetate 4.91 ± 1.09 8.26 ± 0.49 3.00 ± 0.60
Ethyl nonanoate 0.17 ± 0.04 0.00 ± 0.00 0.16 ± 0.05
Ethyl 3-phenylpropionate 0.07 ± 0.04 0.00 ± 0.00 0.00 ± 0.00
9-Hexadecenoic acid, ethyl ester 1.25 ± 0.52 0.91 ± 0.14 0.00 ± 0.00
Ethyl caprate 3.25 ± 0.36 1.97 ± 0.38 4.70 ± 0.24
Ethyl myristate 0.39 ± 0.06 0.36 ± 0.13 0.41 ± 0.23
Ethyl palmitate 1.35 ± 0.55 2.19 ± 1.13 1.09 ± 0.12
Subtotal 31.35 22.89 30.80
Other esters
Butanoic acid, 4-hydroxy- 0.00 ± 0.00 0.07 ± 0.04 0.00 ± 0.00
Carbonochloridic acid, octyl ester 1.74 ± 0.45 0.00 ± 0.00 0.00 ± 0.00
Methyl benzoate 0.54 ± 0.11 0.75 ± 0.22 0.95 ± 0.12
Ethyl isopentyl succinate 0.28 ± 0.08 0.00 ± 0.00 0.21 ± 0.09
Isopropyl palmitate 0.00 ± 0.00 0.00 ± 0.00 0.61 ± 0.32
Subtotal 2.56 0.82 1.78
Acohols
3-(Methylthio)propanol 0.11 ± 0.05 0.11 ± 0.04 0.00 ± 0.00
Benzyl alcohol 0.15 ± 0.04 0.11 ± 0.03 0.14 ± 0.05
1-Octanol 1.86 ± 0.25 0.00 ± 0.00 0.00 ± 0.00
Phenethyl alcohol 28.84 ± 2.20 47.16 ± 11.42 25.95 ± 7.16
6-Octen-1-ol,3,7-dimethyl-, (3R)- 0.22 ± 0.06 0.17 ± 0.03 0.21 ± 0.08
Decyl alcohol 0.13 ± 0.02 0.00 ± 0.00 0.08 ± 0.02
1-Pentadecanol 0.16 ± 0.06 0.07 ± 0.04 0.00 ± 0.00
(±)Dihydrafarnesol 0.15 ± 0.04 0.18 ± 0.05 0.21 ± 0.03
Subtotal 31.62 47.79 26.60
Acids
Octanoic acid 3.87 ± 0.63 0.00 ± 0.00 3.31 ± 0.82
Nonanoic acid 0.37 ± 0.12 0.31 ± 0.30 1.51 ± 1.15
Decanoic acid 1.60 ± 0.09 0.91 ± 0.10 4.79 ± 0.63
Lauric acid 0.11 ± 0.05 0.18 ± 0.05 0.18 ± 0.07
Tetradecanoic acid-1-13c 0.09 ± 0.06 0.14 ± 0.08 0.00 ± 0.00
Palmitic acid 2.15 ± 1.07 2.69 ± 0.60 0.00 ± 0.00
Oleic acid 0.47 ± 0.10 1.50 ± 1.06 1.44 ± 1.24
Subtotal 8.66 5.73 11.22
Volatile furans and phenols
3-Methyl-4-isopropylphenol 0.17 ± 0.07 0.07 ± 0.05 0.26 ± 0.09
4-Hydroxy-3-methoxystyrene 1.00 ± 0.70 0.46 ± 0.11 0.33 ± 0.06
2,6-Di-tert-butyl-4-methylphenol 0.13 ± 0.02 0.14 ± 0.03 0.12 ± 0.04
2,4-Di-tert-butylphenol 0.17 ± 0.05 0.18 ± 0.06 0.14 ± 0.08
3,4-Epoxytetrahydrofuran 0.15 ± 0.08 0.24 ± 0.13 0.00 ± 0.00
Subtotal 1.63 1.09 0.85
Other compounds
Furfural 1.66 ± 0.44 1.86 ± 0.69 2.55 ± 0.29
Benzaldehyde 0.00 ± 0.00 0.27 ± 0.06 0.00 ± 0.00
Phenylacetaldehyde 0.00 ± 0.00 0.09 ± 0.06 0.10 ± 0.03
1-Nonanal 0.23 ± 0.07 0.18 ± 0.09 0.00 ± 0.00
2-Methyltetrahydrothiophen-3-one 0.00 ± 0.00 0.00 ± 0.00 0.33 ± 0.07
Acetophenone 0.24 ± 0.02 0.00 ± 0.00 0.00 ± 0.00
2-Undecanone 0.00 ± 0.00 0.00 ± 0.00 0.10 ± 0.06
Damascenone 0.00 ± 0.00 0.14 ± 0.02 0.00 ± 0.00
Bicyclo[2.2.1]hept-2-ene,1,7,7-trimethyl- 0.05 ± 0.03 0.00 ± 0.00 0.15 ± 0.12
Cyclodecane 0.00 ± 0.00 0.00 ± 0.00 0.12 ± 0.03
Subtotal 2.19 2.54 3.35

fermented by Jiuqu + S. cerevisiae EC1118 was significantly
decreased, indicated a higher safety of the kiwi wine by the
mixed fermentation.
The different metabolites in different samples were
mainly caused by the different microorganism flora.
Jiuqu consisted a mixture of yeasts, molds, and bacteria
(Bora et al., 2016; Lv et al., 2012). Since each kind of
microorganisms had their specific metabolic pathways,
they could generate aboundant substances from the
same source. For example, obligately homofermentative
lactic acid bacteria can only convert hexoses to lactic
acid bacteria through Embden–Meyerhof–Parnas
pathway, but obligately heterofermentative lactic acid
bacteria degrade hexoses by the phosphogluconate
pathway, producing not only lactic acid as the end pro-
duct but also ethanol or acetic acid and carbon dioxide
(Buron-Moles, Chailyan, Dolejs, Forster, & Mikš, 2019).
Therefore, with the help of a rich microbial community,
the produced wine had higher quaily but lower methanol
content.
3.3. Volatile profile
The volatile flavor substances in different kiwi wines were
shown in Figure 3. The detail of identified components for
974 A.-J. CHEN ET AL.

each sample was reported in Figure 4 and Table 4. A total of 4. Conclusion

44 volatiles were detected for the three samples. Jiuqu fer-
In this study, kiwi wine was produced in three different culture
mented wine had higher alcohol content than other groups,
conditions S. cerevisiae EC1118, Jiuqu and Jiuqu + S. cerevisiae
which was constant with the results shown in Table 2, that
EC1118. Mixed fermentation showed high quality of the final
Jiuqu fermented wine showed higher ethanol content. But for
products: it showed lower total organic acids but higher lactic
ethyl esters and acids content, Jiuqu fermented wine was
acid content, lower methanol content and higher sensory
significantly lower than S. cerevisiae EC1118 and Jiuqu +
quality. As S. cerevisiae fermented kiwi wine was not satisfying
S. cerevisiae EC1118 groups. The results indicated that Jiuqu
enough for consumers, the present study provided
could help increase the ethanol production, but cannot
a promising strategy to improve the quality of kiwi wine by
increase the volatile profile of the final products. These results
mixed fermentation of Jiuqu + S. cerevisiae EC1118. Even
also confirmed in the mixed fermentation group, there were
though Jiuqu is designed to ferment rice in China, and the
no significant differences between the wines produced by
microbial composition of Jiuqu is not fully understood yet, the
S. cerevisiae EC1118 alone and Jiuqu + S. cerevisiae EC1118.
results indicated that it is useful to utilize combinations of
microorganisms to optimize the kiwi wine production.
Therefore, it is of great value to explore the microbial combi-
3.4. Sensory evaluation nation which is suitable for kiwi wine in further studies.
Friedman Test was carried out and the results of ranking are
mentioned in Table 5. The results of the Friedman Test were
Disclosure statement
F = 20.033, and F’ = 20.372. According to the order of rank
and rank sum of Friedman test approximate threshold table, No potential conflict of interest was reported by the authors.
the threshold of J, P, α (15, 3, 0.05) was 6.400. The results
showed significant difference in the three kinds of wines (F’ Funding
> 6.400). At the significance level of 5%, the least significant
difference (LSD) was 10.37; however, the absolute value of This work was supported by Science and Technology Achievement
Conversion Project of Sichuan Province [Grant No. 2017CC0067],
the rank sum difference of three kinds of wines was greater
Transformation of Technical Achievements in the Production of
than 10.37. Therefore, the conclusion that the quality of Kiwifruit Brandy.
three kinds of wines had significant differences among
each other could be drawn (p < 0.05). Particularly, the sum
rank of kiwi wine fermented by Jiuqu + S. cerevisiae EC1118 ORCID
was 42.5, which was better than that of kiwi wine fermented An-Jun Chen http://orcid.org/0000-0002-8933-6998
by yeast S. cerevisiae EC118 or Jiuqu alone (p < 0.05). The
contents of ethyl ester, organic acids, alcohol, volatile furan
and phenol of kiwi wine fermented by S. cerevisiae EC1118 References
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