Food and Nutrition Sciences, 2015, 6, 935-943

Published Online July 2015 in SciRes. http://www.scirp.org/journal/fns

http://dx.doi.org/10.4236/fns.2015.610097

Feasibility of Using Corncob as the Substrate

for Natural Vinegar Fermentation with
Physicochemical Changes during the
Acetification Process
Kaustav Chakraborty, Joyjit Saha, Utpal Raychaudhuri, Runu Chakraborty*
Department of Food Technology and Biochemical Engineering, Jadavpur University, Kolkata, India
Email: *[email protected]

Received 15 July 2014; accepted 28 July 2015; published 31 July 2015

Copyright © 2015 by authors and Scientific Research Publishing Inc.

This work is licensed under the Creative Commons Attribution International License (CC BY).
http://creativecommons.org/licenses/by/4.0/

Abstract
Vinegar made from corncob, an agricultural waste product of corn, is of interest in terms of its po-
tential as a new functional condiment with physiological characteristics derived from polyphenols.
The advent of food out of waste using lignocellulosic substrates is a modern biotechnological ap-
proach to enhance sustainability. The fermentation parameters for production of fermented
vinegar from corncobs were optimized. The ethanolic fermentation was completed by Saccharo-
myces cerevisiae in 52 hours at 36˚C producing 3.16% (v/v) ethanol. The acetic acid fermentation
was carried out by Acetobacter aceti in 48 hours at 34˚C for producing vinegar having 3.91% (w/v)
acetic acid. Total phenolic compound of corncob wine and vinegar was reported as 49 mg/100ml
and 43 mg/100ml respectively. Total flavonoid content was determined to be about 12 mg/100ml
of corncob wine and 9 mg/100ml of corncob vinegar. Corncob wine and vinegar showed highest
antioxidant activity with 39% and 37.94% respectively. The structural change during the two step
fermentation was also confirmed by scanning electron microscope (SEM). The Hunter color value
of the corn cob vinegar was also evaluated. With the current trend of shifting to natural products,
corncob vinegar with better health benefits and economic significance, certainly scores high above
its synthetic counterpart.

Keywords
Corncob, Vinegar, Antioxidant, Color, SEM

*
Corresponding author.

How to cite this paper: Chakraborty, K., Saha, J., Raychaudhuri, U. and Chakraborty, R. (2015) Feasibility of Using Corncob
as the Substrate for Natural Vinegar Fermentation with Physicochemical Changes during the Acetification Process. Food
and Nutrition Sciences, 6, 935-943. http://dx.doi.org/10.4236/fns.2015.610097
K. Chakraborty et al.

1. Introduction
Vinegar, a weak form of acetic acid, is a versatile product that has been used in all corners of the world for over
thousands of years as acidic seasoning by virtue of its physiological effects. Its acclivity in terms of use makes
its study worthwhile. One of the prime qualities of vinegar to reduce pH aids in disinfection of food, processing
equipment, neutralizes smell and prevents growth of pathogenic bacteria like Escherichia coli in food. It was
also found that addition of vinegar to rice reduced glycemic index of rice by 20% - 35% [1]. Natural vinegar is
best known for its health values since ages. The vinegar available in the market is mostly synthetic that lacks
any food or medicinal value. Natural vinegar could be obtained from different raw materials having good fer-
mentation potential. Vinegar produced from different natural substrate have unique potential like plum vinegar
have high antioxidant potential than red wine, black vinegar from brown rice have dihydrosinapic acid and rice
shochu distilled vinegar shows strong LOOi-scavenging activity. Natural vinegar has long been used as a sea-
soning and preserving agent and is produced by a fermentation process from wine, apple juice, honey, rice,
malted barley etc. In north eastern regions of Asia, such as China and Japan, vinegar is traditionally produced
from cereals, primarily rice. Cereal or starch based source—natural vinegar is obtained by a three stage fermen-
tation process [2]. Among the vinegar produced, alcohol vinegar accounts for the largest percentage, which is
produced from diluted purified ethanol, but the interest for natural wine vinegars is now growing among the
consumers. However, alternative processes have been extensively explored because of the high production cost
associated with the utilization of natural fruit sources. Vinegar production from agricultural wastes could be a
possible source because it has the potential to realize cheaper production of vinegar with low environmental im-
pact by the effective utilization of renewable resources such as agricultural wastes. Among various agricultural
wastes, corncobs are regarded as a promising agricultural resource because corn is widely cultivated and grown
as choicest crop especially for its huge consumer acceptability and proteinaceous vegetable source. Utilization
of waste in wine and vinegar production would go a long way in contributing considerably to the economy of
emerging natural vinegar market. Corn is one of the major crops of India. The production of corn in India was
12,000,000 Mt against the total world production of 602,026,822 Mt [3]. It is mainly used to produce glucose,
fructose, high fructose corn syrups and other products. Corn cobs are mainly used as a fuel in furnaces or used
as manure in the soil. The method of fermentation process has been identified as an economically viable option
for conversion of these substrates to fermentation products, namely ethanol and xylitol. The conversion of glu-
cose and xylose to ethanol by culture has been successfully obtained using a Saccharomyces cerevisiae. In the
present study, projecting corncob vinegar as a suitable method for waste utilization economically, needs re-
search in screening of variety of corn, suitable yeast strain and standardization of fermentation methods. The
quality characteristics of the Corncob wine (CCW) and Corncob vinegar (CCV) were monitored during fermen-
tation by measuring pH, color and acidity. Of particular interest was the ability to retain antioxidant properties
throughout the fermentation process. Total phenolic and flavonoid content and antioxidant (DPPH) properties of
the Corncob wine and vinegar obtained were also profiled along with structural feasibility for fermentation and
acetification by Scanning Electron Microscope (SEM).

2. Material and Methods

2.1. Chemicals and Instruments
Dextrose, KH2PO4, K2HPO4, MgSO4∙7H2O, FeSO4∙7H2O, Urea, (Merck, India) were used. Yeast extract, Pep-
tone (Himedia, India), 2,2-Diphenyl-1 picryl—hydrazyl (DPPH) (Himedia, India), Gallic acid (SD fine Chem
Ltd India), Folin-Ciocalteu’s phenol reagent (Merck India), AlCl3 (Merck India), NaNO2 (Merck India) were
used. A Level 1 pH-meter with combined pH electrode (Elico, LI 120) with buffers of pH 3.0 and 7.0 was em-
ployed for the pH of corncob wine and vinegar samples. All the spectrophotometric analyses were implemented
using a spectrophotometer (Hitachi U-2000). Quartz cuvettes with optical length of 10 mm were used. The fol-
lowing samples were subjected to research: Corncob wine (CCW) and Corncob vinegar (CCV). The corn cob
was procured as agricultural waste from West Bengal, India.

2.2. Alcoholic Fermentation

Corn cob was washed with water and cut into pieces. They were pulped using mixer. After the above treatments,
30 g of the pulp sample and 70 ml of distilled water were dispensed in 500 ml conical flasks, plugged with cotton

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and placed on a rotary shaker for intimate mixing. The treated pulp sample obtained in this manner was then
subjected to fermentation using stock culture of wine yeast Saccharomyces cerivisiae (NCIM 3315) procured
from National Chemical Laboratory (NCL) Pune, India. The culture media consisted of 1.0 glucose, 0.3 malt
extract, 0.3 yeast extract and 0.5 peptone all in g/100mL. For the organisms to grow, the temperature was kept at
30˚C, pH 6.5 with a incubation period of 45 hours. The alcohol obtained was subjected for analysis of alcohol
content using GC analysis. 5 mL of CCW sample was centrifuged (Remi C-24, Mumbai, India) at 3500 rpm for
10 minutes. Gas Chromatography of supernatant was used to determine the ethanol concentration. Gas chroma-
tography (GC) analysis was carried out using Agilent Technologies: GC system—7890A gas chromatography,
column—Agilent JK WDB-624 with column ID—250 μm, length—60 m and film length—1.4 μm. The chro-
matograph was fitted with column (28 × 31 × 16 cm) with column gas flow rate 1.4615 ml/min. Helium was
used as a carrier gas and the solvent used is dimethyl sulfoxide (DMSO). The percentage of composition of the
ethanol was calculated by the GC peak areas.

2.3. Acetous Fermentation

After fermentation, acetification was carried on using Acetobacter aceti to produce vinegar. Acetobacter aceti
(NCIM 2116) was accomplished by inoculating on liquid medium, having the composition: 1.0 tryptone, 1.0
yeast extract, 1.0 glucose, 1.0 calcium carbonate, 2.0 agar g/100ml and incubating at 30˚C for 48 h at pH 6.0
with shaking for use as the vinegar starter in corncob wine after alcoholic fermentation. Acetic acid concentra-
tion formed by acetic fermentation was quantified by a HPLC system (JASCO, MD 2015 Plus, Multiwave
length Detector) equipped with absorbance detectors set to 210 nm. The column (ODS-3) was eluted with 0.01
(N) H2SO4 as the mobile phase at a flow rate of 0.5 mL/min and a sample injection volume of 20 μL. Standard
acetic acid (Merck, India) was used as an external standard.

2.4. Total Phenol Content (TPC)

The total phenolic content (TPC) for CCW and CCV were quantified using the Folin-Ciocalteu (FC) reagent
was measured according to Di Stefano and Singleton method [4] [5]. A 150 µL of Folin-Ciocatlteu reagent with
aliquot of 20 µL samples was taken in a spectrophotometer cuvette, 600 µL of a 15% Na2CO3 solution and dis-
tilled water were added to make the final volume of 3000 µL. The increase in absorbance was measured at 784
nm. After 2 hours, the concentration of TPC was expressed as mg/100 ml gallic acid equivalent, determined by a
calibration curve graph. Folin-Ciocalteu reagent could also be reactive with other chemicals in wine and vinegar,
including thiols, inorganic ions, vitamin derivatives as well as amino acids. Therefore, although the result of Folin-
Ciocalteu method represents a rough approximation of total phenolic content in most cases, compounds other
than polyphenols need to be considered.

2.5. Total Flavonoid Content

The total flavonoid concentration was measured according to the method of [6]. Firstly 5.0% 75 μL of NaNO2
was added, shortly followed by 10% 150 μL of AlCl3∙6H2O and left for 5 min. After adding 500 μL of NaOH,
the absorbance was measured at 510 nm. The total flavonoid content of samples was measured using a mg cate-
chin equivalent of 100 mL.

2.6. DPPH Radical Scavenging Activity

The effect of the extracted sample on DPPH radical was estimated according to the procedure described by
Brand-William method [7]. The sample extract (0.1 mL) was added to 3.9 mL of DPPH (100 μM) in ethanol that
was prepared daily. After incubation time of 45 min the absorbance was determined at 515 nm. Ethanol solution
of 0.1 mL was used as blank and 3.9 mL of DPPH solution as control. The inhibitory percentage of DPPH was
calculated according to the following Equation (1):
1 − ( absorbancesample absorbancecontrol )  × 100
DPPH scavenging effect ( % ) =
 

2.7. Color Value

Hunter colorimeter (Lab Scan XE, Hunter Associate Laboratories, Inc., Reston, VA) was used for the color

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K. Chakraborty et al.

measurements. By using illuminant C and 2˚ observer angle, the sample was placed in a 1-cm path length optical
glass cell in total transmission mode. The color parameters recorded were: (L*), a* value, b* value, chroma
(saturation, C*) and hue angle (h˚).

2.8. Scanning Electron Microscopy

For the microscopic studies, samples of corncob and vinegar were freeze dried using a Lyophilizerand then
transferred to a desiccator. After the samples were dehydrated, they were mounted over the stubs withdouble-
sided conductivity tape and were observed using a scanning electron microscope (model FEI Quanta -200 MK2)
[8] at a specific magnifications of 800× and 3000×.

3. Results and Discussion

3.1. Factors Regulating the Production of Ethanol and Vinegar
The fermentation parameters for production of fermented vinegar from corncobs were optimized. The ethanolic
fermentation was completed by Saccharomyces cerevisiae in 52 hours at 36˚C producing 3.16% (v/v) ethanol.
The acetic acid fermentation was carried out by Acetobacter aceti in 48 hours at 34˚C for producing vinegar
having 3.91% (w/v) acetic acid. Glycolitic production of ethanol from glucose and fructose affects the subse-
quent acetic oxidation in different ways. In this study, the alcohol content of wine was analyzed using GC and
found to be about 3.16% (Table 1) which was suitable for the acetic acid bacteria to produce acetic acid has
been reported to be responsible for yeast-acetic acid bacteria antagonism in vinegar production [9]. However, in
this case the alcohol content in vinegar was measured and found to be about 2.01%; the drop in the alcohol con-
tent suggests the effective utilization of the sugar by the wine yeast due to alcoholic fermentation. On the con-
trary, acetic acid production can also be inhibited by low ethanol levels also due to increase of incomplete oxi-
dation of polyols and sugars, mainly gluconic acid synthesis via direct glucose oxidation especially at low pH
values [10]. The vinegar had a significant decrease of 57% which was more as compared to the reported 50%
decrease of alcohol content in the jujube vinegar [11]. The drop in the alcohol content could due to conversion
of alcohol content to acetic acid; the acetic acid (3.19%) confirms the conversion [11]. The acids that may be
present mainly in the wine are tartaric acid, tartarate, malic acid and potassium hydrogen mallate [12]. The in-
crease in acidity in the vinegar was mainly due to acetic acid and other organic acids like citric acid, tartaric acid,
succinic acid, lactic acid and gluconic acid etc. in minor amounts produced by Acetobacter aceti during acetifi-
cation process during the study of the Balsamic and other vinegars [13]. Weak lipophilic acids with low pH
posses a synergic effect which reduces intracellular pH of yeast below the normal physiological values, thereby
inhibiting the yeast growth. So the pH was closely monitored having the mean values of 4.12 and 3.14 in wine
and vinegar respectively, shown in Table 1. As the titrable acidity increases, acetic acid seems to directly affect
enzymatic activities, such as enolase, a key enzyme of glycolysis. The pH was found to be higher than the bal-
samic vinegar (pH—2.2 - 2.6) [13]. The decrease in pH in the vinegar was mainly due to increase in titrable ac-
ids like acetic acid and other organic acids. Fermentation temperature is an important parameter affecting yeast
growth rate. The temperature in this study was kept at 30˚C as yeasts are mesophilic organisms with an optimum
growing temperature included between 20˚C and 40˚C. When temperature decreases below 20˚C in a wine fer-
mentation, ethanol tolerance and growth rate of S. cerevisiae are greatly inhibited [14]. Exothermic metabolic
reactions during the full-fermentation phase may raise the temperature above 40˚C, thus reducing the yeast vital-
ity and ethanol yields causing undesirable stuck fermentations. So a constant monitoring was done to restrict the
temperature to 30˚C - 32˚C.
In most cases of acetous fermentation, it is associated with a decrease in the parameters as far as studies till

Table 1. General composition of CCW and CCV.

Alcohol Acetic acid Total phenols Total flavonoids

Sample pH DPPH %
(%v/v) (g, %) (mg/100mL) (mg/100mL)
CCW 3.16 ND 4.12 49 12 39

CCV 2.01 3.91 3.14 43 9 37.94

ND: not detected. All values are mean of three replications.

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 K. Chakraborty et al.

date are considered. The individualistic reason for decrease has been provided below but one of the key factors
of acetification changes, common to all parameters, is contributed by the type of container used in the acetifica-
tion [15].

3.2. Total Phenolic Content

Polyphenols are the most abundant phytochemicals and characterization of the phenolic profile of vinegar is
very important since the phenolic profiles of vinegars are responsible not only for their sensory characteristics
but also for most of the health benefits attributed to them due to its antioxidant activity. Corncob wine and vine-
gar have been reported to have total polyphenol content of 49 mg/100mL and 43 mg/100mL respectively. How-
ever, it is the amount of extractable polyphenols as the portion of polyphenols bound to cell wall or trapped
within the food matrix could not be determined [16]. The antioxidant activity of phenolic compounds is related
to the hydroxyl groups linked to the aromatic ring, which are capable of donating hydrogen atoms with electrons
and stabilizing free radicals. Recently, various health-promoting effects of vinegar like suppression of cardio-
vascular diseases and prevention of appetite increase along with increase in digestion absorption have been re-
ported. These effects are assumed to be due to the phytochemicals in vinegar drinks, such as organic acid, amino
acids, and phenolic compounds. Among them, polyphenol has high antioxidant activity [13]. The total polyphe-
nol content of CCV is found to be higher than vinegars of Japanese apricot, apple and brown rice vinegar that
have been reported to have relatively low polyphenol content below 25 mg/100mL [17] and red ginseng vinegar
(38.06 mg/100mL) [18]. But on the other hand the following vinegars: balsamic vinegar (372.53 mg/100mL),
blackberry vinegar (87.25 mg/100mL), pomegranate vinegar (64.15 mg/100mL), cactus vinegar (48.98 mg/
100mL) and Red persimmon vinegar drink (60 mg/100mL) [18] had high amount of total polyphenol content
than CCV. There are more than a thousand types of polyphenols among other products like green tea etc. But-
principle compounds related to the vinegar phenolic content are gallic acid, a naturally abundant plant com-
pound, derivatives and esters of benzoic acid, tartaric acid and cinnamic acid along with metabolic products of
yeast, such as tyrosol [19]. CCW had a phenolic content of 49 mg/100mL which decreased in the CCV to 43
mg/100mL. The decrease in the phenolic content was in concord with the decrease in the total phenol content by
acetification process which was highest for cider vinegar (40%) and lower for red and white wine vinegars (13%
and 8%) respectively [12]. The result reported in our study was about 12% which was alike of the red wine
vinegars. The decrease in total phenolic content can be attributed to polymerisation or condensation reactions
with other phenols, a similar phenomenon in vinous and berry substrates [20]. Accordingly CCV showed a de-
crease in the total phenolics content but the percent decrease was lower than cider vinegar and higher than white
wine vinegars.

3.3. Total Flavonoid Content

Flavonoid is found in plant pigments that have flavones as a basic structure and occurs in plant leaves, roots,
fruits, and stems. Flavonoids generally contain anthoxanthin, anthocyanin and catechin having antibacterial,
anti-allergic and anti-inflammatory activities. Corncob wine and vinegar had a flavonoid content of 12 mg/
100mL and 9 mg/100mL respectively. The content of flavonoids in corncob wine and vinegar is less correlated
to their antioxidant capacity and has lower ratio flavonoids: polyphenols. It is similar with other vinegars like
balsamic. Therefore, other compounds such as phenolic compounds different from flavonoids and Maillard re-
action products, contribute to their antioxidant capacity. The flavonoid content of corncob vinegar was similar to
9.23 mg/100mL found in the red ginseng vinegar and higher than cactus vinegar having flavonoid content below
5 mg/100mL. But the value is relatively low compared to fruit vinegars such as blackberry vinegar (51.12 mg/
100mL) and others [18].

3.4. Antioxidant Activity

The DPPH free radical scavenging assay has been widely used to evaluate antioxidant capacities. Antioxidant
properties of compounds present in corncob wine and vinegar are important as they retard or prevent oxidation
process. The DPPH capacity of CCW and CCV are indicated in Table 1. Corn cob wine and vinegar obtained
had a yellowish appearance. The antioxidant activity of CCW and CCV were evaluated using DPPH free radi-
cal-scavenging assays. In the DPPH assay, the DPPH of CCW and CCV were found to be 39% and 37.94%, re-
spectively. For production of vinegar using inoculated wines rather than spontaneous acetification process, was

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K. Chakraborty et al.

a contributing factor for a low antioxidant activity and high phenolic content in vinegar. This step was necessi-
tated to maintain flavor profile by high phenolic content. However, the result was in accordance with previously
reported results in the literature where the antioxidant activity of blueberry wine was higher than blueberry
vinegar [21]. After acetic fermentation, the antioxidant activity of CCV was 37.94%, which was higher than the
one of balsamic vinegar [22]. Therefore, it could be concluded that the antioxidant activities of CCW and CCV
though low, but was comparable to other wines and vinegars. The DPPH scavenging activity of the CCV and
CCW was credited to the cumulative presence of higher concentration of polyphenols and other bioactive com-
pounds. The result shows that vinegars are also free radical-scavengers, particularly of the peroxyl radicals,
which are the major propagators of the oxidation chain of fat, thereby terminating the chain reaction [23]. The
CCV showed high scavenging capacity higher than that of vinegars prepared pomegranate vinegar (5.63%) and
blackberry vinegar (12.07%) [18]. However the reported radical-scavenging activity on DPPH radicals was less
than that of the persimmon vinegars made from persimmon Saijo which showed the highest (84.2% ± 1.2%) but
higher scavenging activity than vinegars from polished rice and apple which were found to be 13.5% and 11.2%,
respectively [24]. In our study, the vinegar was produced in laboratory scale involving glass vessel which usu-
ally displayed the lowest values for all the parameters than those produced in wood and oak barrels. Vinegars
produced in oak and cherry barrel had highest amount of total phenols and antioxidants respectively as the stable
anthocyanin-derived compounds are formed due to oxygen permeation through the wood thereby decreasing
monomeric anthocyanins [15]. So the values reported in our study would certainly be enhanced if produced in
pilot plants involving oak and cheery wood barrels.

3.5. Hunter Color Parameters

Color is an important aspect of wine and vinegar quality because of its relevance in sensory analysis and as an
indicator of oxidation and aging processes. Rosy vinegar, for its special color and flavor, is very popular in the
Southeast area of China [25]. Every winery producing high-quality vinegar standardizes its color so that it may
serve to identify its brand. The coordinates L*, a*, b* determines the wine and vinegar color. At this scale, L*
measures luminosity that varies from zero (black) to 100 (pure white); a* and b* values represent the levels of
tonality and saturation, with +a (indicating red), −a (indicating green), +b (indicating yellow) and –b (indicating
blue). According to the values in Table 2, the CCW and CCV was light (L* value of 32.35 and 32.43 respec-
tively). Both CCW and CCV are visually yellowish green in terms of color with values of a* (−4.19), b* (11.19)
for CCW and a* (−4.20), b* (11.26) for CCV, as observed in this work. C* (Chroma) is a parameter that indi-
cates the contribution of a* (redness) and b* (yellowness), respective values of Chroma for wine and vinegar are
11.94 and 12.01 respectively, with Hue angle (color, h˚) of 69.54˚ and 69.54˚ respectively (Table 2). Overall,
the CCW and CCV had a close approach to brewed vinegar, cider vinegar and brown rice vinegar [26]. The
color of the wine and vinegar are reportedly same and color differences are small enough not to be noticed by
eye (Table 2). As a final step in the production of sherry wine vinegars, color is adjusted before bottling by the
addition of a small quantity of caramel, but however in our case it was not done. There is reportedly a strong
correlation with the colour and antioxidant activity. Antioxidant activity of white wines to be markedly lower
compared to the activity of red wines [27]. It was further confirmed that wines from berries being soft with a
thin skin as well as light in color like cloudberries and strawberries had practically low antioxidant activity than
from the wines from berries with strong color and relatively small size with a higher proportion of tough skin
such as black currants, cranberries and crowberries, shown to exert high antioxidant activity [28]. With the yel-
lowish green colour it is shown to have comparably high antioxidant activity as that of white wine vinegars.

3.6. Scanning Electron Microscope (SEM) Studies on Structure Suitability of the Substrate
Scanning electron microscopy of the corn cobs (in raw forms), before the cells adhesion (Figure 1(a) and

Table 2. Hunter color values of CCW and CCV.

Sample L* a* b* Chroma (C) Hue angle

CCW 32.35 −4.19 11.19 11.94 69.54˚
CCV 32.43 −4.20 11.26 12.01 69.54˚
Chroma (C) = [(a*) 2 + (b*) 2]1/2 and hue angle (h˚) = arctan (b*/a*). Values presented are mean of duplicate analysis.

940
 K. Chakraborty et al.

Figure 1. (a) SEM analysis of raw corncob substrate; (b) SEM analysis of raw corncob substrate (closer view); (c) SEM
analysis of acetified corncob substrate fermented corncob substrate (Corncob vinegar); (d) SEM analysis of acetified corncob
substrate fermented corncob substrate (Corncob vinegar) (closer view).

Figure 1(b)), revealed that fermentation and acetification did not occur in a homogeneous form on the material
structure, but it was more favoured in specific region especially in rough and porous structures. In fact, such
structures allow microorganisms, in this case yeast, to attach more firmly to the surface than the smooth struc-
tures. This phenomenon has also been reported in immobilization studies where corn cob was used successfully
as a support material for the production of ethanol [29]. The yeast cells were attached on the materials surface
by adhesion due to presence of a large number of cavities (Figure 1(a) and Figure 1(b)), which provided a
natural entrapment of the cells (Figure 1(c) and Figure 1(d)), a phenomenon that is preferred in the beverage
production. In brief, corn cobs were the most suitable support materials for adhesion of S. cerevisiae cells during
fermentation. This may be due to the greater availability of nutrients (sugars and nitrogenous compounds) near
the surface of the support. In addition, the greater nutrient availability in the cavities of is also an attraction to
yeast cells, which encouraged yeast cells to migrate into the inner parenchyma cells during the process [30]. The
appearance of the yeast cells was observed in the medium after fermentation and acetification (Figure 1(c) and
Figure 1(d)). Adhesion of S. cerevisiae is essentially dependent upon electrostatic interactions between the
support and the normally negatively charged cell surface and the attachment of the yeast cells on the support
may have occurred as a result of hydrogen bonding, entrapment of the cells in corncob pieces, and the van der
waal forces [30]. Because of this, corncob pieces are believed to have natural entrapment of yeast cells into the
porous structure of said materials and due to physical adsorption by electrostatic forces between the cell mem-
brane and the yeast cells. Similar results have been observed in yeast cells when using in apple pieces for wine
preparations [8]. Therefore the structure of corncob is an important advantage considering the economy of the
process and aids in more bioconversion of the masses to produce ethanol and acetic acid.

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4. Conclusion
Corncob vinegar was produced and its quality characteristics along with phytochemical contents were monitored
during fermentation. During wine fermentation, highest alcohol yield was found to be 3.16% and during acetic
acid fermentation, acetic acid content was estimated to be 3.91%. From wine fermentation to acetification, there
was considerable change in alcohol content from 3.16% to 2.01% with a little change in pH. Vinegar produced
from inoculated wine showed total phenolic content and antioxidant activity comparable to other wine vinegars.
Physiochemical changes were assayed and found that all the parameters were decreasing from wine to vinegar
involving total phenols decreasing from 49 mg/100mL to 43 mg/100mL, antioxidants decreasing from 39% to
37.94% and the flavonoids from 12 mg/100mL to 9 mg/100mL. The substrate feasibility was established by mi-
croscopic studies which entails an important factor for yeast and acetobacter fermentation. The results in this
research indicate the possibility that corncob vinegar could be used as a new product with antioxidant activity
that is comparable to other vinegar products.

Acknowledgements
This research work is financially supported by the “UGC-BSR Research Fellowship in Science for Meritorious
Student” under University Grants Commission (UGC), India.

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