INTRODUCTION

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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

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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.

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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)

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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

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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;

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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

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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

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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

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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

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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

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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,

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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,

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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;

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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.

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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

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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

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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