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Production of Herbal Wine from Hibiscus rosa-sinenesis by Using Different

Strains of Saccharomyces cerevisae

Article in Advances in Life Sciences · January 2016

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9434Advances in Life Sciences 5(20), Print : ISSN 2278Advances-3849, 9434inLife-9437,Sciences20165(20), 2016
Production of Herbal Wine from Hibiscus rosa-sinenesis by Using Different Strains
of Saccharomyces cerevisae
SNEHLATA TIWARI, SANGEETA SHUKLA, SONIA MORYA AND KAUSHAL KISHOR
Warner School of Food and Dairy Technology SHIATS, Allahabad, Uttar Pradesh
ABSTRACT
The diversity and the composition of the yeast
micropopulation significantly contribute to the sensory
characteristics of wine. The growth of each wine yeast
species is characterized by a specific metabolic activity,
which determines concentrations of flavour compounds
in the final wine. Effect of different yeast strains of
Saccharomyces cerevisiae i.e. MTCC178, MTCC 180,
MTCC 786 Strains on the enological and sensory
characteristics of Hibiscus wines was determined.
However, it must be underlined that, within each
species, significant strain variability has been recorded.
The wide use of starter cultures, mainly applied to
reduce the risk of spoilage and unpredictable changes of
wine flavour, can ensure a balanced wine flavour, but it
may also cause a loss of characteristic aroma and
flavour determinants. Sensory evaluation scores of
different wines revealed that the must fermented with
the yeast strain MTCC 178 had higher scores for colour,
taste, flavour and overall acceptability. Thus, the
beneficial contribution from the yeast increases when
starter cultures for winemaking are selected on the basis
of scientifically verified characteristics.
Key words Saccharomyces cerevisiae, MTCC178,
MTCC 180, MTCC 786, Hibiscus wines.
The consumption of a variety of local herbs and
vegetables by man is believed to contribute significantly to
the improvement of human health, in terms of prevention or
cure of diseases because plants have long served as a useful
and rational source of therapeutic agents
The Hibiscus rosa-sinensis belongs to the family
Malvaceae. Traditionally flower can be used as anti
asthamatic agents. many chemical constitute such as
cyanidin, quercitin, hentriacontane, calcium oxalate,
thiamin, riboflavin, niacin, and antimicrobial substances are
present in this flower (Zhao J, 2010). The petal have some
protective mechanism against microbial attack in most of
the parts. The H. rosa-sinensis flower petal of large number
of plant species growing in the vicinity of our environment
were screened for their antimicrobial activity. the
antioxidant properties of flavonoids, polyphenolic
Compounds and anthocyanins contained in the flower can
prevent the oxidation of Low-Density Lipoproteins (LDL),
which is associated with the disease.
The characterization of wine yeasts of different
species for by-product formation has underlined that the
yeast species itself is a prominent factor in determining the
wine composition (Houtman et al., 1980; Benda, 1982;
Herraiz et al., 1990; Romano, 1997; Brandolini et al., 2002).
In particular, the characterization of S. cerevisiae, the
principal actor of the winemaking, has revealed that, in
addition to the production of ethanol, this yeast generates
many secondary metabolites that are key determinants of
wine quality (Fleet, 1990; Lema et al., 1996; Lambrechts
and Pretorius, 2000; Fleet and Heard, 1993).
Wine flavour is composed by a wide variety of
compounds with different aromatic properties. It includes
flavour compounds originating from the grapes (varietal
flavour), compounds formed during operations of extraction
and conditioning of must (prefermentative flavour), other
compounds produced by yeasts and bacteria during
alcoholic and malolactic fermentation (fermentative
flavour) and compounds that appear during the ageing
process (post-fermentative flavour), as reviewed by Schreier
(1979), Boulton et al. (1995), and Rapp (1998). Volatiles
identified in wines are usually dominated by fermentation
products, since these compounds are present in the highest
concentrations. Therefore, conversion of grape sugars to
alcohol and other end-products by specific yeast
populations may yield wines with distinct organoleptic
quality. The various yeast species and strains that develop
during the overall fermentative process metabolise grape
juice constituents, principally the sugars, to a wide range of
volatile and non-volatile end-products, which influence and
determine the types and concentrations of many products
that contribute to the aroma and flavour characteristics of
the wine. The major volatile products of yeast metabolism,
ethanol and carbon dioxide, make a relatively small
contribution to wine flavour. Conversely, organic acids,
higher alcohols and esters and to a lesser extent
acetaldehyde constitute the main group of compounds that
form the ‘‘fermentation bouquet’’ (Rapp and Versini, 1991).
When present in excess concentrations, these compounds
may also be regarded as undesirable. Here we report by-
product formation in a large number of natural wine yeasts
in order to emphasize the function of yeast species and
strains in wine flavour.
MATERIALS AND METHODS
The present study was conducted in the research
laboratory of Warner School of Food & Dairy Technology,
Sam Higginbottom Institute of Agriculture, Technology and
Sciences (Deemed University), Allahabad.
The material and methods were adopted during this
investigation are given below -
 Hibiscus rosa-sinensis flowers were collected from
Agronomy department of SHIATS, Allahabad.
 Sugar, KMS, Di-ammonium phosphate were
purchased from local market of Allahabad
 Different types of Saccharomyces cerevisiae strains
MTCC180, MTCC 178, MTCC 786 were collected
from IMTECH, Chandigarh, India.
TIWARI et al., Production of Herbal Wine from Hibiscus rosa-sinenesis by Using Different Strains of Saccharomyces cerevisae 9435

Dried Crushed flower of H. rosa-sinensis

0
Extraction of flower ( boiling at 79.5 C for 10 mins.)

Filtration

0
In Filtrate addition of sugar (24 Brix)
+
Addition of KMS (150 ppm)
+
Di-ammonium phosphate(0.3 gm/L).

Keeping for an hour

Addition of different strains of S. cerevisiae (@ 2%

MTCC178 MTCC180 MTCC 786

Fermentation (for 10 days at room temperature)

Filtration

Maturation (at room temperature)

Bottling

0
Pasteurization (at 82 C for 20 mins.)

Cooling

Storage

Flow Chart for making Herbal wine

Maintenance of yeast culture fermentation process. An amount of 2 ml of the yeast was

Three yeast strains namely Saccharomyces cerevisiae
MTCC178, S. cerevisiae MTCC 180, S.cerevisiae MTCC
786,were procured from institute of Microbial Type Culture
Collection, IMTECH, Chandigarh, India. All the cultures
were grown at 300 c under agitation(150 rpm) for 24h in
yeast extract peptone dextrose broth (YEPD) medium and
adjusted to pH 5.0. All the strains were sub-cultured on
YEPD(1% (w/v) yeast extract 2% (w/v) peptone, 2% (w/v)
dextrose; agar slants at fortnight intervals and stored in
refrigerator at 4±0 C, until further use.
Fermentation of must prepared from Hibiscus rosa-
sinensis
Preparation of Must
The Hibiscus petals were manually sorted and washed
in cold water to remove dirt. About 80 gm of the hibiscus
petals powder boiled in 1000 ml of water at a starting
temperature of 800 C for 10 minutes.
Fermentation of Must
The extracts obtained were then ameliorated or
enhanced, using of table sugar to raise the sugar level to
24° Brix, which is in the range of sugar levels tolerable
by most yeast for alcohol production (Bisson, 2001).
Brewer’s yeast Saccharomyces cerevisiae was used in the
added for each 1 litre of must. Sulfur dioxide was added to
the must in the form of sodium metabisulphite at a
concentration of 0.4 g/l to stabilize the must. The must were
poured into three different sterile plastic fermenting
containers and seeded aseptically with 2 ml yeast culture
and stirred for even distribution of the yeast. A vent was
made on the lid and stuffed with sterile cotton wool to allow
for the exchange of gases. The musts were subjected to
primary fermentation at 28 ± 2°C for eight days to produce
the fermented must. The yeasts were filtered out to obtain
the young wine.
Clarification of Young Wine
Each type of the young wines were fined using
bentonite. Bentonite was added slowly without foaming to
the wine while stirring and allowed to stand for ten days
before filtering out using cheese cloth (Dharmadhikari,
2000a; Zoecklein et al., 1990).
Bottle Ageing of Unclarified and Clarified Fermented
Must
The unclarified and clarified fermented musts were
bottled in one litre transparent glass bottles and well corked.
Bottle aging was allowed for twenty weeks to allow the
development of characteristic flavour of the wines
(Dharmadhikari, 2000).
9436 Advances in Life Sciences 5(20), 2016

Table 1. Average mean values analyzed for various quality attributes of Herbal wine

Treatments
Parameters MTCC 178 MTCC 180 MTCC786
Colour & Appearance 8.30 7.90 8.24
Flavour & Aroma 8.70 8.06 7.76
Consistency 8.84 8.36 7.74
Overall Acceptability 8.32 7.73 7.13

Racking of Wine
The wines were racked off the lees every five weeks.
This was done by the use of a sucker pipette to avoid
contamination and ensure minimum oxygen contact with the
wine. Storage Conditions of Unclarified and Clarified
Fermented Must The bottled unclarified and clarified fermented
musts were kept in a cool dry cabinet at an average temperature
of 28oC with no disturbances for twenty weeks.
Sensory evaluation
The sensory evaluation of wines of different
treatments was conducted by a trained panel of 10 judges.
Each judge was given a set of wines separately in isolated
booths and provided with a glass of fresh water to rinse their
mouth before tasting the next sample. Each sample was
evaluated for various quality attributes, viz. colour,
consistency, aroma, overall acceptability, etc. as per the Fig. 1. Average scores in various quality attributes of
prescribed performa. Judges were asked to rate the samples Herbal wine
on a prescribed sensory evaluation performa with earlier
stated attributes. samples were assessed organoleptically
using a 9-point hedonic scale, where 9 correspond to “like CONCLUSION
extremely” and 1 corresponds to “dislike extremely” The wine produced from H. rosa-sinensis has been found
Statistical analysis to be acceptable, as well as meeting all the standards required
All the assays were carried out in triplicates. The by a good wine in terms of color, flavour, taste, aroma and
experimental results were expressed as mean ± standard overall acceptability for a bouquet wines. H. rosa-sinensis is
deviation. The data were analyzed using one way analysis readily available and it is cheap, thus it can be a good substrate
of variance (ANOVA) . Differences were determined using for wine industry. The use of the Hibiscus flower for wine can
Fisher’s Protected Least Significant Difference (LSD) with be exploited commercially as this would lead to conservation of
foreign exchange. Equally, more studies are needed that would
P ≤ 0.05.
compare quality of the H. rosa-sinensis wine aged for a long
RESULTS & DISCUSSION period of time with commercial wines from other sources. It
Herbal Wine production sounds common but the can be concluded that various yeast strains influenced the
novelty of this work is the use of Hibiscus flower and juice sensory characteristics of the resulting wines. Sensory
which is fermented using a different yeast strains of evaluation scores of different wines revealed that the must
Saccharomyces cerevisiae purchased from IMTECH fermented with the yeast strain MTCC 178 had higher scores
chandigarh for wine production. Below are the findings. for colour, taste, flavour and overall acceptability. All the wines
More recently, wine makers and wine researchers have were of commercial acceptability. It is concluded that the use of
come to realize that Saccharomyces yeasts contribute in appropriate yeast strain for the preparation of Hibiscus wine is
significant measure to the flavour and quality of wine than very important along with other vinification practices as it
previously thought (Romano et al., 1997a). for determining influenced the sensorial quality of wine.
their potential effects on the organoleptic qualities of the final
product. The differences in the composition of wines made ACKNOWLEDGEMENTS
from different yeast species appear to be quantitative rather
I would like to thank prof. (Dr.) Ramesh Chandra
than qualitative (Romano, 1997).The products of fermentation
Dean, Warner school of Food and Dairy technology, Sam
are usually identical, but the relative amounts of the various
higginbottom institute of agriculture technology and
compounds differ between different yeast species. From the
sciences (deemed university), Allahabad, for providing
sensory analysis result shown in Table-1 that there was
guidance and all require facilities and thank to My Advisor
significant difference (P > 0.05) among those three wines in
Dr. Sangeeta Shukla assistant professor, WSFDT, for her
terms of colour, flavor taste and overall acceptability. MTCC
constant co-operation, help, guidance and support during
178 wine is rated higher than MTCC 180 & MTCC 786 wine in
project period.
all attributes
 TIWARI et al., Production of Herbal Wine from Hibiscus rosa-sinenesis by Using Different Strains of Saccharomyces cerevisae 9437
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Received on 17-10-2016 Accepted on 22-10-2016