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1. INTRODUCTION
Fermentation is the oldest form of preservation, essentially consisting of
transformation of the simple raw materials into value added products by utilizing the
phenomenon of the growth of microorganisms or their activities on various substrates.
As a science, the food fermentation has an element of biological sciences especially
the microbiology, genetics and biochemistry, as a technology the food technology,
chemical engineering along with integral component of sciences involved in food
toxicity, acceptability and food nutrition. The term fermentation is derived from the
Latin word ‘Fervere - to boil’ thus describing the actions of yeasts on extracts of fruits
or malted grain. Later it was Pasteur in 1856, who discovered the microbial
association of fermentation. Traditionally, fermented foods and beverages are known
to human being since 5000 years. But scientifically fermentation process was first
applied to wine fermentation signifying a gentle bubbling or boiling condition due to
production of carbon dioxide, during breakdown of sugar or it could be without
liberation of gases (Pedersen, 1979; Dirar, 1993).
Fermentation is a method of preservation through formation of acidulants,
alcohols, antibacterial compounds and improving digestibility of protein and
carbohydrates thus improving the nutritional strength and flavour. Besides this
fermentation also helps to reduce or eliminate antinutrients, natural toxicants, cooking
time and fuel requirements. Fermented foods and beverages enhance the pleasure of
eating and their nutritional roles include indirect contributions through subjective
enhancement of appetite. Fermented foods have several health benefits. It aids in
digestion, promote healthy flora in digestive tract, produce beneficial enzymes, offer
better nutrition and help to absorb vitamins and minerals more effectively from foods.
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Some fermented foods regulate the level of acidity in the digestive tract, also act as
antioxidants and provide anti-carcinogenic effect. (Parvez et al., 2006)
In Indian context, fermented foods confines to mainly milk, cereal based foods
and alcoholic beverages (Lewis and Johar, 1953; Veen et al., 1967; Reddy and
Salunkhe, 1980; Reddy et al., 1982; Salunkhe et al., 1985; Joshi et al., 1990;
Varadaraj et al., 1993; Deshpande and Jha, 2008; Sarkar, 2008; Riat and Sadana,
2009; Kumbhar et al., 2010; Khetra et al., 2011), while globally, wider application
include vegetables, cereals, fruits, meats and fish (Hesseltine, 1965; Ebine 1971;
Banigo and Muller, 1972; Sheih and Beuchat, 1982; Svanberg and Sandberg, 1988;
Haggblade and Holzapfel, 1989; Uzogara et al., 1990; Hugas et al., 1993; Mwesigye
and Okurut, 1995; Iwuoha and Eke, 1996; Runge et al., 1996; Jay,1996; Sankaran,
1998; Girado and Mazas, 2000; Katz, 2001; Ishikawa et al., 2003; Lee and Lucey,
2004; Montano et al., 2004; McKinley, 2005; Wen et al., 2006; Panda et al., 2007;
Yucel and Uren, 2008; Cai, 2010; Fu et al., 2010; Harlow and Phillips, 2011; Mousavi
et al., 2011; Wegkamp et al., 2011). The impact of fermentation process on the
formation of nutrients, both qualitatively and quantitatively has received very less
attention. The knowledge as well as information on nutritional strength and its
implications, increased shelf life of products create an awareness in the consumers
and the scientific evaluation of products through clinical trials creates scientific
evidence and acceptance. Such attempts in the research on fermentation of foods are
welcome for health progression of human beings.
The fermentation process may be struck fermentation, malolactic fermentation
or alcoholic fermentation; where a partial fermentation occur in the first type while
lactic acid bacteria is utilized in the second type and yeast is most commonly used in
alcoholic fermentation. While conducting specific fermentation, efforts are made to
3
control the type of microorganism and the environmental conditions to produce the
desired product. As a result of fermentation, important class of foods termed as
fermented foods are produced. Depending upon the regional tastes and likings, there
are quite large variations in methods of preparation of fermented foods.
The scope of food fermentation ranges from producing alcoholic beverages,
fermented meat, milk and vegetable products to genetic engineering producing super
foods to carry out efficient fermentation to treatment and utilization of waste and
overall producing nutritious and safe products with appealing qualities. (Lim, 1991;
Bhutani and Joshi, 1995; Joshi and Bhutani 1995). It is especially true in certain
regions or countries of the world and in earlier times when bountiful harvest yield
larger quantity of fresh foods than the population could consume (Mukerjee, 1987).
For such foods, fermentation proved its place and is a potent tool to prevent the
postharvest losses. The most common fermented products were rice wine, sugar cane
wine, pickles, fish sauce, fermented noodles, tempeh and oncom. (Lim, 1991). The
preservative effect of fermentation has been due to the formation of ethanol, carbon
dioxide, specific antimicrobial substances like bacteriocins, reutrin etc. and the major
effect is the lowering of pH such as by lactic or acetic acid bacteria; as illustrated by
the bread making and alcoholic beverages and the yeast fermentation results in the
formation of carbon-di-oxide. The preservative effect is more pronounced in the sour
breads where besides yeast, lactic and acetic acid bacteria is the causative factor for
producing acid (Reed, 1981). Many fermented foods have good keeping qualities and
may be kept without refrigeration for periods longer than the fresh unfermented foods.
But like any other food, the fermented foods are also subjected to normal spoilage and
to prevent this, one or more combinations of the preservative techniques are applied
such as low temperature storage, thermal processing or addition of preservatives.
4
Production of fermented foods
Fermented foods and beverages are known to human being 5000 years back.
Egyptians were skilled in the art of preparing fermented beverages and leavened bread
in 3200 B C. The global market value of food fermentation based food industry
estimated to be around 250 million US dollars. The marketed fermented products
show that alcoholic beverages and cheese are the major products. Interestingly, more
than 10% of this accounts for alcoholic beverages (Fig 1) with projected increase of
US $ 27 billion in 1989 to 40 billion in 1990 (Knorr, 1995) as observed by Houwink
(1984), “ Moneywise biotechnology is mainly beer and wine”.
Fig 1: World markets for some of the food fermenation derived products
Source: Joshi and Pandey, 2004
But to-day the fermentation based foods has gained much importance and
research is in progress. The global scenario on the fermented products is given in
Table 1 and they are based on soybean, barley, milk, fish, meat, rice, grape and a few
traditional fruits (Oener et al., 1993; Jun et al. 1999). Ham et al. (1997) developed a
beverage drink based on mountain edible herbs. Lee et al. (2002) studied the sensory
effect of traditional Korean medicinal rice wines brewed with functional herbal
powders or extracts. Kirova and Ionkova (1999) studied the influence of some herbs
on the activity of the microbial lactic starter for Bulgarian yoghurt.
0 5000
10000 15000 20000 25000
US $ (million)
5
Table 1: Oriental fermented foods
Product and Country / region
Substrate Microorganism(s) involved
Nature and use Reference
A. Cereal based fermented foods Ang-kak, China Rice Monascus purpures Dry red powder used as
colourant Palo et al., 1960; Hesseltine, 1965
Bagni, Caucasus Darassum, Injera, Mongolia Ethiopia
Millet Millet Wheat, barley, teff, maize etc.
Candida guilliermondii Liquid drink Liquid drink Bread substitute
Stewart and Getachew, 1962; Oda et al., 1983; Chavan and Kadam, 1989; Zegeye, 1997.
Jalebies, India, Nepal, Pakistan
Wheat flour Saccharomyces bayanus Syrup filled confectionery Riat and Sadana, 2009
Kanji, India
Rice and carrots Hansenula anomala Sour liquid added to vegetables
Kamla et al., 2001
Kenkey, Ghana Maize - Steamed eaten with vegetables
McKay and Baldwin, 1990; Halm et al., 1993; Jespersen et al., 1994
Lao-chao, China, Indonesia
Rice Rhizopus oryzae Rhizopus chinensis Chalmydomucor oryzae, Saccharomyces species
Soft, glutinous eaten with vegetables
Wang and Hesseltine, 1970
Mahewu, South Africa Merissa, Sudan
Maize Sorghum
Lactic acid bacteria Saccharomyces species
Liquid drink Liquid drink
Chavan and Kadam, 1989; Odunfa et al., 2001
Minchin, China Wheat gluten Paecilomyces species Aspergillus species Clasdosporium species Fusarium syncephalastrum
Solid condiment Wood, 1977
6
Miso, Japan, China
Rice and soybean
Aspergillus oryzae Torulopsis etchelisii Saccharomyces rouxii
Paste, soup base Hesseltine, 1965; Ebine 1971; Beuchat, 1978a; Beuchat, 1978b; Sheih and Beuchat, 1982
Nan, India, Pakistan, Afghanistan, Iran
Unbleached wheat flour
Saccharomyces cerevisiae Lactic acid bacteria
Solid snack food Riat and Sadana, 2009
Ogi Nigeria, West Africa
Maize Lactic acid bacteria Cephalosporium species Fusarium species Aspergillus species Penicillium species Saccharomyces cerevisiae
Paste, staple breakfast food
Akinrele, 1964
Puto Philippines
Rice Lactic acid bacteria, Saccharomyces cerevisiae
Solid snack food Sanchez, 1975; Sanchez 1977
Torani India
Rice Hansenula anomala Candida guilliermondii Candida tropicals Geotrichum. candidum
Seasoning for vegetables Padmaja and George; 1999
B. Legume based fermented foods Chee-fan, China Soybean whey,
curd Mucor species Aspergillus glaucus
Solid, eaten fresh like cheese
Padmaja and George; 1999
Dawadawa, West Africa, Nigeria
African locust bean
Lactic acid bacteria, yeast
Solid eaten fresh or in stews
Reddy et al. 1982; Beuchat, 1983
Hamanatto, Japan Soybeans, wheat flour
Aspergillus oryzae Streptococcus species Pediococcus species
Raisin like flavouring agent for meat or fish or eaten as a snack
Yokotsuka, 1991
Kecap, Indonesia & nearby regions
Soybeans, wheat
Aspergillus oryzae Lactobacillus species Hansenula species
Liquid, condiment and seasoning agent
Sardjono and Sudarmadji, 1992; Roeling et al., 1994; Roling et al., 1999;
7
Saccharomyces species Kenima, Nepal Sikkim and Darjeeling
Soybeans Solid snack food Reddy et al. 1982
Ketjap, Indonesia Black soybeans Aspergillus oryzae
Syrup, seasoning agent Wood, 1977
Khaman, India Bengalgram - Solid, cake like breakfast food
Rajalakshmi and Vanaja, 1967; Ramakrishnan et al., 1976; Lakshmi, 1978
Meitauza, China, Taiwan
Soybean cake Actinomucor elegans Solid, fried in oil or cooked with vegetables
Zhu et al., 2008;
Meju, Korea
Soybeans Aspergillus oryzae Rhizopus species
Paste, seasoning agent Joo et al., 1992; Kwon and Kim, 2005
Natto, Northern Japan
Soybeans Bacillus natto Solid cake, as a meat substitute
Hesseltine, 1965; Wang and Hesseltine, 1970; Kiuchi et al., 1976; Reddy et al., 1982
Ontjom, Indonesia Peanut cake intermediate
Neurospora Solid, roasted or fried Beuchat, 1976; Beuchat, 1978a; Beuchat, 1978b; Veen et al., 1968
Bongkrek, Central Java
Coconut press cake
Rhizopus oligosporus Roasted of fried in oil, meat substitute
Cox et al., 1993; Garcia et al., 1999
Papadam, India Blackgram Saccharomyces species Solid, crisp condiment Lancaster et al. 1982 Soybean milk Soy sauce, China, Japan, Philippines and Oriental countries
Soybeans and wheat flour
Lactic acid bacteria, Aspergillus oryzae Aspergillus sojae Lactobacillus species Saccharomyces rouxii
Liquid drink, Liquid seasoning agent for meat, fish and cereals
Horitsu et al., 1990; Horitsu et al., 1991; Takenawa and Ise, 1998; Chumchuere and Robinson, 1999; Chun et al., 2008
Sufu, China, Taiwan
Soybean whey and curd
Aspergillus elegans, Micrixalus silvaticus
Solid, soybean cake, condiment
Wai, 1968; Wang and Hesseltine, 1970
8
Mucor substilissimus Tempeh, Indonesia and nearby regions
Soybeans Rhizopus species Fried in oil, roasted, as a meat substitute
Hesseltine and Wang, 1967, Steinkraus, 1978
Warries, India Blackgram flour Candida species Saccharomyces species
Spongy, spicy condiment Reddy et al., 1982
C. Mixed fermented foods Banku, Ghana Maize and
cassava Lactic acid bacteria Used as a staple food Ansah et al., 1987
Burukutu, Savannah regions of Nigeria
Sorghum and cassava
Lactic acid bacteria Candida species Saccharomyces cerevisiae
Liquid creamy drink Faparusi et al., 1973
Dhokla, India
Bengalgram and wheat
Leuconostoc mesenteroides Streptococcus faecalis Trichosporon candida Trichosporon Pullulans
Spongy condiment Reddy et al., 1982; Salunkhe et al., 1985
Dosai, India
Blackgram and rice
Yeasts Leuconostoc mesenteroides
Spongy breakfast food Battacharya and Bhat,1997
Idli India Blackgram and rice
Leuconostoc mesenteroides Trichosporon candida Trichosporon pullulans
Spongy, moist breakfast food
Lewis and Johar, 1953; Veen et al., 1967; Reddy and Salunkhe, 1980; Reddy et al., 1982
D. Tuber crops based fermented foods Chickwangue Congo Gari West Africa
Cassava roots Corynebacterium manihot, Geotrichum candidum, Lactobacillus plantarum, Leuconostoc mesenteroides. Candida species
Pasta, staple food, granular wet paste eaten as a staple with stews
Nwankwo et al., 1989; Okafor and Ejifor, 1990; Cereda and Mattos, 1996; Sokari and Karibo, 1996; Osho and Dashiell, 2002; Oboh et al., 2002
Lafun West Africa &
Cassava roots Bacillus subtilis, Lactobacillus,
Paste, staple food Ijabadeniyi, 2007; Nwabueze and Odunsi, 2007
9
Nigeria Leuconostoc, Streptococcus, Candida
Fufu, Africa
Cassava roots Lactobacillus species Leuconostoc species Saccharomyces cerevisiae
Paste, eaten with soup, sauce or stew
Akindahunsi and Oboh, 1999; Fagbemi and Ijah, 2006
Poi, Hawaii
Taro corms Lactobacillus species Candida vini Geotrichum candidum
Semi solid dish taken with fish or meat
Achi and Akubor, 2000; Huang et al., 2003; Brown et al., 2005
Tape, Indonesia and nearby
Cassava roots or rice
Saccharomyces cerevisiae, Hansenula anomala, Chalmydomucor oryzae, Mucor species, Endomycopsis fibuliger
Soft, solid eaten as a staple
Beuchat, 1983
E. Fermented fish products Bagoong, Philippines
Fish - Paste seasoning agent Veen, 1953
Fish sauce, South east Asia
Fish Bacteria Liquid seasoning agent Uchida et al., 2005
Katsuobushi, Japan Whole fish Aspergillus glaucus Solid dry, seasoning agent Graikoski, 1973 Prahoc, Cambodia
Fish - Paste, seasoning agent Sochivi and Viryak, 2011
F. Fermented milk products Acidophilus Milk, Australia
Cow’s milk Lactobacillus acidophilus Fermented milk drink Jay,1996
Cultured butter milk, Scandinavian and European
Cow’s or buffalo’s milk
Lactococcus lactis. Leuconostoc dextranicum Leuconostoc citrovorum
Fluid with suitable viscosity, of typical clean refreshing acid taste with pleasant flavor and aroma.
Shakila, 2012
10
Dahi, India, Persia
Cow’s or buffalo’s milk
Lactococcus lactis, Streptococcus salivarius, Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus bulgaricus, lactose fermenting yeasts, Mixed culture (not defined)
Fermented milk Varadaraj et al., 1993
Kefir Caucasus
Sheep’s, cow’s, goat’s mixed milk, fermentation in skin bag or in wooden barrels
Lactococcus lactis, Lactobacillus delbrueckii, Saccharomyces kefir, Torula kefir, micrococci, spore forming bacilli
Fermented milk drink with an alcohol
Jay,1996
Kumiss Asiatic steppes
Mare’s, or camel’s milk fermentation in skin bag
Lactobacillus delbrueckii, Lactobacillus bulgaricus, Lactobacillus acidophilus, Torula kumiss, Saccharomyces lactis, micrococci, spore forming bacilli lactis, micrococci, spore forming bacilli
Fermented milk drink with an alcohol
Jay,1996
Lassi India
Cow’s or buffalo’s milk
Streptococcus salivarius Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus bulgaricus
A fermented milk consumed as a beverage after dilution with water
Sarkar, 2008; Kumbhar et al., 2010; Khetra et al., 2011
Leben, Labneh Lebanon and Arab
Goat’s or sheep’s milk, fermentation in skin bag/
Lactococcus lactis, Streptococcus salivarius, Streptococcus thermophilus, Lactobacillus delbrueckii,
Sandwich spread Kadamany, et al. 2002
11
earthenware Lactobacillus bulgaricus lactose fermenting yeasts.
Shrikhand (chakka) India
Cow’s or buffalo’s milk
Streptococcus salivarius, Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus bulgaricus
Sweetened milk product Derived from chakka – partly Dehydrated yoghurt)
Sarkar, 2008; Deshpande and Jha, 2008
Yoghurt (bioyoghurt) Middle Asia, Balkans
Cow’s milk, goat’s or mixed milk
Streptococcus salivarius, Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus bulgaricus, Micrococcus and other lactic acid cocci, yeasts, molds
Drink, dietary Supplements for infant consumption
McKinley, 2005; Katz, 2001; Lee and Lucey, 2004
G. Fermented meat products Chorizo Spanish
Pork Lactobacillus species Lactobacillus sake Lactobacillus plantarum
Sausage Rovira et al., 1997
Salami Europe
Pork Lactobacillus, Micrococcus
Sausage Campanini et al., 1993
Sausage Central Europe, U.S.A
Mammalian meat, generally pork and/or beef, less often poultry
Lactic acid bacteria (lactobacilli, pediococci), Catalase positive cocci (Staphylococcus carnosus,) Staphylococcus xylosus, Micrococcus varians) sometimes yeasts and/or moulds
Ground meat with a skin around it
Hugas et al., 1993; Lucke, 1998
H. Fermented fruits and vegetable products Burong mustala Mustard Lactobacillus brevis, Salad, side dish Lee, 1994; Rhee et al., 2011
12
Philippines Pediococcus cerevisiae Dakguadong Thailand
Mustard leaf Lactobacillus plantarum Salad, salt side dish Lee, 1994; Rhee et al., 2011
Dhamuoi Vietnam
Cabbage, various vegetables
Leuconostoc mesenteroides, Lactobacillus plantarum
Salad, side dish Lee, 1994; Rhee et al., 2011
Fermented Fruit Beverages International
Peach, melon, grape, watermelon, cashew apple, pomegranate, pineapple, banana orange, papaya, passionflower fruit
Bifidobacterium, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus delbrueckii
Liquid drink Beng et al., 1994; Runge et al., 1996; Wei et al., 2009; Chanprasartsuk et al., 2010; Fu et al., 2010; Wegkamp et al., 2011; Mestry et al., 2011; Kretschmer and Moll, 2011. Pereira et al., 2011; Mousavi et al., 2011;
Gundruk, Nepal Mixed vegetables
Lactobacillus plantarum, Pediococcus pentosaceus
Dried vegetable product Tamang et al., 1988
Kimchi, Korea, East Asia
Korean cabbage, radish, various vegetables, red pepper, mustard leaf
Leuconostoc mesenteroides, Lactobacillus brevis, Lactobacillus plantarum
Fermented in brine Kim et al., 1999; Pyo et al., 2000; Sim and Han, 2008; Han et al., 2009; Han et al., 2011; Cho, 2010; Lee and Lee, 2010;
Pickled fruit and vegetables, Indian sub-continent South East Asia East Asia
Cucumber, lemon, mango, ginger, red pepper, Japanese radish,
Lactobacillus plantarum, lactic acid bacteria, Leuconostoc species, Lactobacillus casei, Leuconostoc mesenteroides
Pickle Girado and Mazas, 2000; Ishikawa et al., 2003; Ozcelik and Ulu, 2002; Montano et al., 2004; Wen et al., 2006; Panda et al., 2007; Yucel and Uren,
13
Africa Europe
onion, garlic, carrot, Sweet potato, cabbage, mustard, red turnip, in broccoli, cauliflower, cabbage
2008; Panda et al., 2009; Sosinska and Obiedzinski, 2011;
Sauerkraut, International
Cabbage,
Leuconostoc mesenteroides, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei
Used in salads, consumed raw or cooked with meat or sausages
Kuensch et al., 1992; Yu et al., 2001; Kristek et al., 2004; Kusznierewicz et al., 2008; Champagne et al., 2009; Wiander and Korhonen 2011
Vinegar, International
Blueberries, Ginger, kiwi, orange, orange, apple, mango, passion fruit, carrot, tomato, pomegranate, pumpkin
Yeast , Acetobacter Cooking Hur and Lee, 1998; Yae et al., 2007; Su and Chien, 2007; Zhang, 2008; Sahara et al., 2010; Marques et al., 2010
I. Beverages – alcoholic and non-alcoholic Beer, whisky, International
Barley, rye, corn, wheat, rice, sorghum, finger millet
Lactobacillus species, yeasts ,moulds and Bacillus species
Alcoholic drink Banigo and Muller, 1972; Svanberg and Sandberg, 1988; Haggblade and Holzapfel, 1989; Uzogara et al., 1990; Mwesigye and Okurut, 1995; Iwuoha and Eke, 1996; Sankaran, 1998; Steinkraus,
14
1998; Cai, 2010; Harlow and Phillips, 2011;
Coffee, Cocoa, Tea International
Coffee berries, coffee beans, tea leaves, cocoa bean
Lactic acid bacteria, yeasts and acetic acid bacteria
Non Alcoholic drink, powder
Anon, 1992; Kenya Coffee Research Foundation, 1993; Woelore, 1993; Kunz and Murdia, 1994; Fuchs, 1995; Ludwig and Arnold, 1995; Avallone et al., 2001; Cooper, 2006; Oda et al., 2006; Anon, 2008; Camu et al., 2008; Dario and Eskes, 2009; Campos et al., 2011
Wine International
Fruits, rice Saccharomyces cerevisiae, Schizosaccharomyces acidodevoratus, Saccharomyces uvarum, Oenococcus oeni, Lactobacillus plantarum, Leuconostoc oenos
Alcoholic drink Kishkovskaya and Abdullabekova, 1990; Joshi et al., 1990; Edwards et al., 1990; Pena et al., 2005; Cascales et al., 2005; Sacchi et al., 2005; Bayon et al., 2005; Muratore et al., 2007; Fan et al., 2009; Woo et al., 2010; Jeon and Lee, 2011;
15
Development of South African herbal tea was reported by Toit et al. (1998).
Oener et al. (1993) reported about a traditional fermented food “Tarhana” from the
Middle East and Turkey based on yoghurt, mint and spices. Lee et al. (1998) studied
the effect of mixed medicinal herb extracts with antimicrobial activity on the shelf life
of kimchi. Fermented health foods have been developed using oat, corn, maize,
cowpea, rice, wheat, legume, finger millet etc. (Akapunam and Sefadedeh, 1995;
Sharma and Khetarpaul, 1998; Olukoya et al., 2000; Bhatia and Khetarpaul 2001,
Basappa, 2002; Martensson et al., 2002 and 2003). Fermented beverage has been
developed based on custard apple, carrot, orange, maize, jamun, banana etc. (Shukla
et al., 1991; Zulu et al., 1997; Berry, 1998; Rodrigues et al., 2002). Fermented foods
and beverages enhance the pleasure of eating and their nutritional roles include
indirect contributions through subjective enhancement of appetite.
Though, vegetable fermentation has received the attention of researchers, the
vegetable juices, beverages fermentation have seen a limited approach. The
processing as fermented vegetables, (Mukerjee, 1987; Kuensch et al., 1992;
Gerdauskene et al.,1992; Hozova et al., 1993; Kuchta et al., 1994; Nabais and
Malcata, 1995; Lee and Cho, 1996; Pragna and Tanvi, 1997; Uma et al.,1998; Savard
et al. , 2000; Nagy et al., 2002; Viander et al., 2003; Lee et al., 2004; Yunoki et al.,
2004; Roberts and Kidd, 2005; Sim and Han, 2008; Xiaoyang et al., 2009) fruits and
vegetables combination as well as pickling of vegetables (Girado and Mazas, 2000;
Ishikawa et al., 2003; Ozcelik and Ulu, 2002; Montano et al., 2004; Wen et al., 2006;
Panda et al., 2007; Yucel and Uren, 2008; Panda et al., 2009; Sosinska and
Obiedzinski, 2011) are quite common. Still in India, fermented vegetables usage is
less. The research work carried out on the vegetable juices fermentation is given in
Table 2.
16
Table 2: Fermented vegetable beverages
Substrate Microorganism(s) involved Nutrients Reference Cucumber
Bifidobacterium bifidum, Lactobacillus casei, Lactobacillus. plantarum, Lactococcus diacetylactis, Leuconostoc mesenteroides, Leuconostoc oenos, Pediococcus pentosaceus Propionibacterium shermanii Propionibacterium pentosaceus
Not reported Chavasit et al., 1991
Green pea and onion
Lactic acid bacteria Not reported Karoviova et al., 1992
Cabbage, red beet, carrot, celery, tomato, hot pepper
Lactic acid bacteria Increased vitamin-C, thiamine, riboflavin, β-carotene
Afanaseva et al., (1992)
Carrot and cabbage
Lactobacillus delbrueckii Lactobacillus plantarum
Not reported Hybenova et al., 1995
Carrot, celery and pumpkin puree
Lactobacillus plantarum Not reported Kraevska et al., 1996
White cabbages and red beets
Lactobacillus plantarum Leuconostoc mesenteroides Lactobacillus plantarum
Not reported Nowakowskaja and Lipowski, 1996
Artichoke tubers
Aspergillus niger Saccharomyces cerevisiae
Not reported Nakamura et al., 1996
Turnip, black carrot, red beet
Lactobacillus plantarum Lactobacillus brevis Bakers' yeast
Not reported Ozler and Kilic, 1996
Red beet Paracoccus denitrificans Not reported Kolb et al., 1997
Beetroot Lactobacillus plantarum, Lactobacillus cellobiosus, Lactobacillus xylosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus buchnerii Streptococcus lactis
Not reported Gavrilova et al., 1997
Cucumber Saccharomyces rosei Not reported Passos et al., 1997
Carrot Bifidobacterium Not reported Park et al., 1997 Red beet and carrot
Lactic acid bacteria Not reported Rasic, 1998
Beets, carrots, cabbages
Dried lactic acid bacteria Increased vitamin-C
Gorenkov et al., 1998
17
Pumpkin Acetobacter Not reported Hur and Lee, 1998
Carrot, cabbage, peas, cucumber, celery and fermented radish
- Not reported Lee et al., 1998
Orange, apple, carrot, capsicum, tomato
- Not reported Stern, 1998
Fruit-vegetable
Lactic acid bacteria strains isolated from kimchi – natural
Not reported Cheigh et al., 1998
Fruit-vegetable
Lactic acid bacteria strains isolated from kimchi - natural
Not reported Kim et al., 1998
Red beet Propionibacteria and lactic acid bacteria
Increased Vitamin B12, folacin
Babuchowski et al., 1999
White cabbage and carrot
Lactobacillus plantarum , Lactobacillus plantarum Lactobacillus delbrueckii
Not reported Karovicova et al., 1999
Radish Natural lactic acid fermentation
Not reported Kim et al., 1999
Onion Saccharomyces cerevisiae Not reported Horiuchi et al., 2000
Carrot, apple, celery, watercress, jujube and lycci
Lactic acid bacteria were isolated from traditionally made dongchimi – natural
Not reported Kim and Choi, 2000
Cabbage, carrot, beet and onion
Lactic acid bacteria Not reported Gardner et al., 2001
Cucumber Lactobacillus plantarum Not reported Lu et al., 2001 Red beet Paracoccus denitrificans Not reported Tomczak and
Czapski, 2002 Cabbage Lactobacillus plantarum
mixed culture of Lactobacillus plantarum and Saccharomyces cerevisiae
Not reported Karovicova et al., 2002
Cabbage and garlic
Lactobacillus plantarum Not reported Kohajdova and Karovicova, 2003a
Sugar beet Lactobacillus acidophilus , Lactobacillus plantarum Lactobacillus delbrueckii
Not reported Klewicka et al., 2004
18
Carrot Lactic acid bacteria Not reported Demir et al., 2004
Tomato Lactic acid bacteria Not reported Kohajdova et al., 2004b
Carrot Leuconostoc mesenteroides Lactobacillus pentosus
Not reported Bergqvist et al., 2005
Beet Lactobacillus plantarum, Lactobacillus delbrueckii, Lactobacillus casei Lactobacillus acidophilus
Not reported Yoon et al., 2005
Cabbage, carrot, celery and beetroot
Lactobacillus plantarum Not reported Karovicova and Kohajdova, 2005
Beetroot and carrot
Brewer’s yeast autolysate Increased vitamin-C
Rakin et al., 2005
Cabbage Lactobacillus plantarum, Lactobacillus casei Lactobacillus delbrueckii
Not reported Yoon et al., 2006
Red beetroot Lactic acid bacteria Not reported Czyzowska et al., 2006
Carrot Lactic acid bacteria Not reported Demir, 2006 Cabbage, tomato, pumpkin and courgette
Lactobacillus plantarum Not reported Kohajdova et al., 2006
Multi-vegetable
Propionibacterium Not reported Radyko et al., 2006
Carrot, apple and tomato
Lactobacillus bulgaricus and Streptococcus thermophilus derived from yoghurt and Lactobacillus plantarum from pickles – natural
Not reported Feng and Ying, 2006
Ginger Yeast Not reported Kuboi, 2006 Beetroot and carrot
Lactobacillus acidophilus brewer's yeast autolysate
Increased vitamins
Rakin et al., 2007
Carrot Commercial starters and isolated lactic acid bacteria strains
Not reported Wiander and Ryhanen, 2007
Cucumber and onion
Lactobacillus plantarum Not reported Kohajdova et al., 2007
Sweet potato Lactobacillus plantarum Not reported Smita et al., 2007
Carrot Bifidobacterium lactis Bifido bifidum strains
Decreased α and β carotenes
Yang, 2007
Celery and beetroot
Bifidobacteria culture Not reported Moraru et al., 2007
Carrot Leuconostoc mesenteroides Not reported Jo et al., 2008 Onion Saccharomyces cerevisiae Not reported Jose-Maria et
al., 2008
19
Carrot and beet
Leuconostoc mesenteroides isolated from carrot juice
Not reported Son et al., 2008
Mixture of vegetables
Lactic acid bacteria Not reported Furuse et al., 2008
Cabbage, radish and celery
Lactic acid bacteria Not reported Buruleanu et al., 2008
Carrot, tomato
Yeast and acetobacter Not reported Zhang, 2008
Carrot Lactobacillus rhamnosus Lactobacillus bulgaricus
Not reported Filomena et al., 2008
Sugar beet Lactic acid bacteria Not reported Visser et al., 2009
Tomato Lactobacillus Streptococcus thermophilus
Not reported Hua et al., 2009
Carrot and amla
- Not reported Sahota et al., 2009
Carrot and beetroot
Lactic acid bacteria Not reported Buruleanu et al., 2009
Onion Lactic acid bacteria isolated from various fruits and vegetables – natural
Not reported Choi et al., 2009
Tomato - Not reported Jang et al., 2010 Tomato Bifidobacterium breve,
Bifidobacterium longum, Bifidobacterium infantis
Not reported Koh et al., 2010
Cabbage, carrots and beetroot
Lactic acid bacteria Not reported Manea and Buruleanu, 2010
Watermelon and carrot
Lactobacillus acidophilus Not reported Mestry et al., 2011
Cucumber, white and red cabbage
Lactobacillus acidophilus Not reported Buruleanu et al., 2012
20
The Table 2 reveals that mainly cucumber, carrot, red beet, pumpkin,
cabbage, radish, ginger, onion and a combination of celery and beetroot; cabbage and
onion; cabbage, carrot, beet and onion; beetroot and carrot; green pea and onion;
carrot, celery and beetroot; carrot, celery and pumpkin have been studied. The main
emphasis is on the microorganisms used. Literature clearly brings out the fact that the
work on nutrient formation and flavour changes has been attempted by very few
researches. Afanaseva et al. (1992) reported that sauerkraut juice had high amounts of
vitamins such as C, β-carotene, thiamine, and riboflavin. Montano et al. (2004)
reported that on a dry basis, the fermented product was found to have a higher content
of riboflavin, α-tocopherol and but a lower thiamine level than the unfermented
product. Flavour component studies were carried out by Pyo et al. (2000) in mustard
leaf kimchies, Ishikawa et al. (2003) in Japanese pickle, Kim et al. (2009) in garlic
and Ku et al. (2010) in ginger, during fermentation process. Secondly, the use of
natural and commercial cultures has been studied by few of the researchers. Natural
culture such as lactic acid bacteria was isolated by kimchi isolates (Cheigh et al.,
1998; Kim et al., 1998), dongchimi isolates (Kim and Choi, 2000), yoghurt and pickle
isolates (Wang and Dong, 2006) and isolated from fruits and vegetables (Choi et al.,
2009). Commercial cultures were used for vegetable fermentation by the researchers
Rakin et al. (2005) to ferment beetroot and carrot; and by Wiander and Rhyanen
(2007) for carrot fermentation. These products are in ready to drink form but may
have a shelf life of few weeks; further preservation of these juices is limited, so
further preservation of these juices needs the attention. In India, the fermentation
process is more adopted for cereal, legume based products, Ayurvedic based
pharmaceutical industries and alcoholic beverages. But the fermentation of the
products leads to better release of nutrients, stability of vitamins, flavors and provide
21
convenience of ready to eat or drink status. Vegetables being good source of vitamins
and minerals, further availability can be improved by fermentation process. In
general, the literature on vegetable fermentation is relatively less as compared to other
staple foods. In India the attempt on vegetable based products is limited. In the recent
years it is gaining the attention of consumers and researchers.
The fermentation leads to preservation of juices for short periods as well
provide protective and nutritive properties to the products (Campbell, 1994). Lactic
acid bacteria synthesizes vitamins (Crittenden et al., 2003) and antimicrobials
(Cleveland et al., 2001) and increase their contents in fermented products. Vegetables
are strongly recommended in the human diet since they are rich in antioxidants,
vitamins, dietary fibres and minerals. The major part of the vegetables consumed in
the human diet are fresh, minimally processed, pasteurized or cooked by boiling in
water or microwaving. Minimally processed and, especially, fresh vegetables have a
very short shelf-life since subjected to rapid microbial spoilage and the above cooking
processes would bring about a number of not always desirable changes in physical
characteristics and chemical composition of vegetables (Zia-ur-Rehman et al., 2003;
Zhang and Hamauzu, 2004). Hence the strategic approach of fermentation of
vegetable juices aids in improving the minor nutrients and this area is still open for
research. The vegetables routinely used in our country especially different types of
gourds, roots, tubers are not attempted at all. Therefore there is a need to develop
vegetable based fermented foods and study their impact on human health benefits.
Therapeutic value of fermented foods
Fruits and vegetables are rich in functional components such as vitamins and
antioxidants. Recently, the use of vegetable materials for the production of dietary
22
beneficial drinks has increased in the food and health industry (Luckow and
Delahunty, 2004). Fruit and vegetable juices could serve as a good medium for
functional ingredients such as probiotics and prebiotics (Shah, 2001; Yoon et al.,
2005). These products are recognised as healthy foods and can be consumed
frequently by a significant percentage of consumers. The investigation of vegetable
juices with synbiotic properties is followed with growing interest by the food
industry, which has high expectations for food products that meet the consumer
demand for a healthy lifestyle. Therefore, obtaining a fermented juice from these
sources requires the use of specially selected cultures to ensure proper progress of the
fermentation process and a guaranteed consistent products with beneficial
organoleptic features. However, very little literature is available on vegetable
fermented beverages.
The present study includes two vegetables-ashgourd and bittergourd for which
commercial yeast and mixed lactic cultures were used as starter cultures for
fermentation. Alcoholic fermentation was adopted for ashgourd fermented juice
(beverage) and lactic acid fermentation was adopted for bittergourd fermented juice
(beverage). The comparison of starter cultures from commercial and natural sources
as well as standard cultures were compared for the functional, nutritional and quality
parameters of the fermented juices (beverages). The optimisation of process
conditions for fermentation such as starter cultures and fermentation period was
achieved by using statistical software, with response surface methodology (RSM).
Response surface methodology is a collection of statistical and mathematical
technique useful for developing, improving and optimizing process (Myers and
Montgomery, 2002). The principles of the approach are explained by Henika (1972,
1982) and Giovanni (1983). It is a designed regression analysis meant to predict the
23
value of a dependent variable based on the controlled values of the independent
variables (Meilgaard et al., 1991; Resurreccion, 1998). The use of RSM in the process
optimization stage leads to the need for an experimental design, which can generate a
lot of samples for consumer evaluation in a short period of time, and thus laboratory
level tests are more efficient (Moskowaitz, 1994). The product optimization time is
greatly reduced from traditional ‘‘cook and look’’ optimization techniques (Rudolph,
2000). From the parameter estimates, it can be determined which variable contributes
the most to the prediction model, thereby allowing the product researcher to focus on
the variables that are most important to the product acceptance (Schutz, 1983).
Ashgourd (Benincasa hispida), a member of the family Cucurbitaceae is one
of the familiar crops that are grown primarily for its use as a vegetable and usually
recognized for its nutritional and medicinal properties especially in Asian countries.
As a rich source of functionally important bioactives and therapeutics such as
triterpenes, phenolics, sterols, glycosides and soluble dietary fiber, the vegetable has
been widely used for therapeutic treatments. (Aslokar et al., 1992; Sivarajan and
Balachandran, 1994; Prachi and Premavalli, 2010). Ashgourd is widely cultivated
throughout the world and stored in the whole form which is made available
throughout the year. In India, ashgourd is used in curries or is coated with sugar or
syrup and eaten as a sweet and recently Majumdar et al. (2009a, 2009b, 2010) have
reported on the stabilization of ashgourd juice and ashgourd blended juice. However
the fermentation process improves the quality in terms of nutrients and flavour.
Ashgourd has several functional properties. It is antimercurial, antidote for alcoholic
poisoning, laxative, diuretic and it can cure internal haemorrhages and constipation
(The Wealth of India, 1962). In the present study, an attempt has made to develop
ashgourd fermented juice (beverage) by alcoholic fermentation using different sources
24
of yeast which is obtained from natural source., i.e. from grape juice and available
commercially., i.e. wet yeast, dry yeast and standard yeast and the study is oriented
towards the analysis of nutritional profile, functional and quality parameters of the
product.
Bittergourd (Momordica charantia L) also known as bittermelon, balsam pear
or karela is widely cultivated as a vegetable and medicinal herb in many Asian
countries and has been shown to exert hypoglycemic effects in animal models and
humans. Bittergourd contains biologically active chemicals that include crude fat,
crude protein, soluble dietary fiber, minerals, essential oil, flavonoids, phenolic acids,
glycolsides, triterpenes. The immature fruits are a good source of vitamin C and also
provide vitamin A (Xie et al., 1998; Braca et al., 2008; Zhang et al., 2009).
Bittergourd can be preserved by dehydration, steeping preservation and pickling and
quite often it is being taken by the diabetic personnel in the form of juice which is
highly bitter. Though it has antidiabetic properties, because of high bitterness, its
utilization is low. The modification of taste, flavour can be achieved by fermentation
process and thus it can be made more palatable. Therefore an attempt is made to
develop bittergourd fermented juice (beverage) using lactic fermentation. Although a
large number of lactic acid bacteria starters are routinely used in dairy, meat and
baked food fermentations, only a few cultures have been used for bittergourd
fermentation (Yan et al., 2006). In the present study, an attempt is made to develop
nutritionally strengthful, highly acceptable, shelf stable bittergourd fermented juice
(beverage) specially suitable for persons with diabetic condition.
However, the studies on nutrient profile have not received much attention thus
the fermentation conditions followed for vegetable juices and the changes in their
nutrients still needs a radical approach to achieve the best quality product.
25
The developed products will be subjected to clinical trials to examine
scientific evidence for health benefits of vegetable fermented juices. Thus the
outcome of the study will be process optimization and their impact on nutrients
formation with established shelf life and their evaluation for physiological benefits.
In India, though a few fermented vegetable products are reported, (Anand and
Dass, 1971; Tamang et al., 1988; Hassan et al., 1993; Joshi et al., 1993;
Balasubrahmanyam and Varadaraj, 1995; Joshi et al., 1996; Kamla et al., 2001; Panda
et al., 2007; Panda et al., 2009) the studies on increased nutritional strength, flavour
etc has not been given importance. Secondly, the present study on vegetable
fermented juices creates the scientific data on nutritional strength and scientific
evidence for its proper utilisation and thus makes the present study an obvious
approach for growth of research.
26
Objectives of the present study
The main objective of the study is to develop the vegetable based fermented
juices, evaluate the nutritional strength and their health benefits. Vegetable gourds are
known for their fibre content, vitamins and medicinal properties. But has a subtle
flavour with different tastes such as ashgourd, bottle gourd, ridge gourd, snake gourd,
have bland taste while bittergourd is highly bitter and pointed gourd is slightly bitter.
The modification of taste and flavour can be achieved by the fermentation process
with increased availability of nutrients. Thus the objective of the study are
1. Studies on development of fermented products using natural
fermentation.
2. Studies on the preservation of fermented products and shelf life
establishment.
3. Studies on the evaluation of the product for acceptance and their health
benefits.
27
Proposed plan of work
1. Studies on the development of fermented ashgourd and bittergourd
juices (beverages): Fermentation process optimization with respect to
time and starter culture.
2. Studies on nutritional changes during fermentation and storage: Changes
in B vitamins, vitamin C, minerals, antioxidants, flavours and total
phenols.
3. Studies on evaluation of fermented ashgourd and bittergourd juice
(beverage) for the health benefits. Sensory acceptance on 9 point
hedonic scale and clinical trials with normal / diabetic subjects as per the
standard protocol