Most of the presently available probiotics foods are based on milk. But there are several problems associated with consumption of milk such as lactose intolerance and increase in the level of the cholesterol level of the consumers. Lactose intolerance and the cholesterol content are two drawbacks related to the consumption of milk and milk products. Currently, products are usually marketed in the form of fermented milk and yoghurt. It has also been suggested that fruit juice could serve as a good medium for cultivating probiotics (MattilaSandholm, 2002). The use of probiotics in the fruit and vegetable juice industry offer to consumers with special needs (vegetarian people with allergic reactions to milk proteins) the possibility to experience the positive effect of probiotic bacteria (Luckow and Delahunty, 2004). There is scanty of information on the lactic acid fermentation of fruits and vegetables juices especially with the respect to optimization of fermentation as probiotic juices and their evaluation as probiotic juices in the light of guidelines for such juices. Similarly, the storage behaviour and safety of such juices have not been documented. Recently, the antioxidant and antimicrobial phenomenon in fermented foods is being advocated, can these juices be designated having these characteristics, is not known (Stevans and Sheldon, 1992; Subrota et al., 2015).
DEVELOPMENT OF PROBIOTIC JUICE BY LACTIC ACID FERMENTATION OF FRUITS AND VEGETABLES Thesis by ANURADHA THAKUR (H-2013-21-M) Submitted to 1985 COLLEGE OF HORTICULTURE Dr YASHWANT SINGH PARMAR UNIVERSITY OF HORTICULTURE AND FORESTRY, NAUNI SOLAN – 173 230 (HP), INDIA in Partial fulfilment of the requirements for the degree of MASTER OF SCIENCE (FOOD TECHNOLOGY) FOOD SCIENCE AND TECHNOLOGY 2015 Dr V K Joshi Retd Professor & Head Department of Food Science and Technology College of Horticulture Dr Y S Parmar University of Horticulture and Forestry, Nauni, Solan – 173 230 CERTIFICATE - I This is to certify that the thesis titled, “Development of probiotic juice by lactic acid fermentation of fruits and vegetables”, submitted in partial fulfillment of the requirements for the award of degree of MASTER OF SCIENCE (FOOD TECHNOLOGY) to Dr Yashwant Singh Parmar University of Horticulture and Forestry, (Nauni) Solan (HP) - 173230 is a bonafide research work carried out by Ms Anuradha Thakur (H-13-21-M) daughter of Shri Madan Singh under my supervision and that no part of this thesis has been submitted for any other degree or diploma The assistance and help received during the course of this investigation have been fully acknowledged Place: Nauni, Solan Dated: July, 2015 (Dr V K Joshi) Chairman Advisory Committee CERTIFICATE - II This is to certify that the thesis titled, “Development of probiotic juice by lactic acid fermentation of fruits and vegetables” submitted by Ms Anuradha Thakur (H-13-21-M) daughter of Shri Madan Singh to Dr Yashwant Singh Parmar University of Horticulture & Forestry, (Nauni), Solan (HP) - 173230 India, in partial fulfilment of the requirements for the degree of MASTER OF SCIENCE FOOD TECHNOLOGY has been approved by the Student’s Advisory Committee after an oral examination of the same in collaboration with the external examiner Dr V K Joshi (Retd Professor & Head) Chairman, Advisory Committee External Examiner Members of Advisory Committee Dr N S Thakur (Professor) Dr (Mrs) Nivedita Sharma (Professor) Dr R K Gupta (Professor) Professor and Head Department of Food Science and Technology Dean’s Nominee Dean College of Horticulture Dr Y S Parmar University of Horticulture and Forestry Nauni, Solan (HP) ACKNOWLEDGEMENTS Well, the list of the people I need to thank will not fit to a single acknowledgement section I just mention some people whose contribution is obvious Firstly, I would like to express my sincere thanks to “Almighty God” The creator of this universe, who has given knowledge to man to think, finds out and explores the world and his creation and without God blessings it was not possible for me to complete this research work, and who permitted me to profound undertake these journey The journey is still continuing with many more destinations to cover At the onset of acknowledging the help of those who have contributed to the realization of this manuscript, I would like to express my profound gratitude and thanks to my esteemed teacher and chairman of my Advisory Committee, Dr VK Joshi, retired Professor and Head of Department of Food Science and Technology for his inspiring vision, valuable suggestions, everlasting patience, logical and necessary criticism, advice and constant encouragement at every step of research work, and finalization of this manuscript It was great opportunity for me to work under his guidance It gives me adnascent pleasure in expressing my heartfelt gratitude to the worthy members of my Advisory Committee: Dr NS Thakur (Professor, Department of Food Science & Technology), Dr (Mrs) Nivedita Sharma (Professor, Microbiology, Department of Basic Science) and Dr RK Gupta(Professor, Statistics, Department of Basic Science) for their valuable suggestions and guidance with their scientific acumen during the investigation and thesis preparation I earnestly acknowledge assiduous effort and sanguine gestures of Dr Anju Dhiman, Prof and Head Dept of Food Science and Technology, Mrs Surekha Attri, Dr KS Thakur, Dr Rakesh Sharma, Dr Devina Vaidya, and Dr Manisha Kaushal and all teachers of my department for their sincere help and valuable suggestions I cordially acknowledge the assistance extended by Smt Hemlata Ji, Smt Shalini Parmar Ji, Sh Tapender Ji, Sh Manglesh Ji, Sh Khemchand Ji, Sh Hemchand Ji, Sh R L Junta Ji, Sh Lekh Ram Ji, Sh Raj Kumar Ji, Smt Kaushalya Ji, Sh Prakash Ji, Sh.Rohit Ji, Sh Narender Ji, Dipti di, Manju di and all staff members of Department of Food Science and Technology Every effort is motivated by an ambition and all ambition has an inspiration behind I owe this place to my ever loving family members, Papa (Sh Madan Singh), Mamma (Smt Mangla Devi), Archana (sister),Vicky (Brother) and Vandu (Bhabhi) for their selfless sacrifice, heartfull blessings, firm faith and supporting me psychologically, emotionally and morally which steered the life of this cipher to the present shape I cant’ forget to mention the care and love showered upon me by Late Sh Kaliram Thakur (Dadaji), Late Smt Manti Devi, all tau ji’s and tai ji’ s, all chacha ji’ s and chachi ji’ s, all my cousins brother especially Devu bhai, Teshu bhai, cousins sisters, all bhabi ji’s, niece and nephew There are certain people in everyone’s life whose best wishes and blessings paved the way towards success and they are one of them I cannot forget to express by the same occasions my deep and sincere thanks to my seniors Vikas sir, Navven sir, Abhimanyu sir, Sangeeta di, Priyanka di, Apoorva di, Nisha di, my classmates Jagriti, Pritika, Hamid, Dharmu sir, Babu, Niku, Abhishek, Aarti and my juniors Mona, Isha, Sheilja and Akriti for their continous support during entire course of my study I have special word of thanks for my friends Monika di, Shilpa, Neha, Rajni, Nitika, Cherry, Jagriti, Ankush, Deepak, Vikku and Late Ritu, who made my every moment living, enjoyable, stress free with their internal support and caring nature, helped me to feel better and encouraged me to carry out my work in time I would like to thank DPT computers for their intense endeavour and kind cooperation in typesetting this manuscript I solely claim all the responsibilities for the shortcomings and limitations in this work Thanks to one and all and to those whose names could not appear but who at one stage or the other has helped me in some ways to achieve the goal Place: Nauni Date: (Anuradha Thakur) CONTENTS Chapter Title Page INTRODUCTION 1-3 REVIEW OF LITERATURE 4-28 MATERIALS AND METHODS 29-40 RESULTS AND DISCUSSION 41-157 SUMMARY AND CONCLUSION 158-164 LITERATURE CITED 165-175 ABSTRACT 176 APPENDICES I-IV BRIEF BIO-DATA LIST OF TABLES Table Title Page 2.1 Physico-chemical composition of apple 2.2 Physico-chemical composition of pear 2.3 Physico-chemical composition of orange 2.4 Physico-chemical composition of cabbage 2.5 Physico-chemical composition of cucumber 2.6 Physico-chemical composition of carrot 10 2.7 Characteristics of lactic acid bacteria 11 2.8 Key genera and species of microbes studied and used as probiotics 14 2.9 Effect of lactic acid fermentation on several diseases 25 4.1 Physico-chemical characteristics of fruits (apple, pear, orange) juices 42 4.2 Physico-chemical characteristics of vegetable (cabbage, cucumber and carrot) juices 43 4.3 Effect of lactic acid fermentation on TSS, pH and titratable acidity of apple juice 46 4.4 Effect of lactic acid fermentation on reducing sugar, total Sugar and total phenols of apple juice Effect of lactic acid fermentation on antioxidant activity and ascorbic acid of apple juice Effect of lactic acid fermentation on viable cell counts, LAB counts of apple juice 48 4.5 4.6 50 52 4.7 Effect of lactic acid fermentation on coliform counts, yeast and mold counts of apple juice 53 4.8 Effect of lactic acid fermentation on antimicrobial activity of apple juice 55 4.9 Comparison of quality of lactic acid bacteria of probiotic apple juices on the basis of physico-chemical parameters 56 4.10 Comparison of quality of lactic acid bacteria of probiotic apple juices on the basis of microbiological quality 57 4.11 Effect of lactic acid fermentation on TSS, pH and titratable acidity of pear juice Effect of lactic acid fermentation on reducing sugar and total sugar of pear juice Effect of lactic acid fermentation on antioxidant activity and ascorbic acid of pear juice 59 4.12 4.13 4.14 Effect of lactic acid fermentation on viable cell counts and LAB counts of pear juice 61 63 64 Table Title Page 4.15 Effect of lactic acid fermentation on coliform counts, yeast and mold of pear juice Effect of lactic acid fermentation on antimicrobial activity of pear juice Comparison of quality of lactic acid bacteria of probiotic pear juices on the basis of physico-chemical parameters Comparison of quality of lactic acid bacteria of probiotic pear juices on the basis of microbiological quality Effect of lactic acid fermentation on TSS, pH and titratable acidity of orange juice Effect of lactic acid fermentation on reducing sugar and total sugar of orange juice Effect of lactic acid fermentation on antioxidant activity and ascorbic acid of orange juice Effect of lactic acid fermentation on viable cell counts and LAB counts of orange juice Effect of lactic acid fermentation on coliform counts, yeast and mold of orange juice Effect of lactic acid fermentation on antimicrobial activity of orange juice Differentiation of quality of lactic acid bacteria of probiotic orange juices on the basis of physico-chemical parameters Differentiation of quality of lactic acid bacteria of probiotic orange juices on the basis of microbiological quality Evaluation of all probiotic fruits (apple, pear and orange ) juices on the basis of rate of fermentation (%/24hr) and rate of acidification (°B/24hr) and lactic acid bacteria counts(108cfu/ml) Effect of lactic acid fermentation on TSS, pH and titratable acidity of cabbage juice Effect of lactic acid fermentation on reducing sugar, total Sugar and total phenols of cabbage juice Effect of lactic acid fermentation on antioxidant activity and ascorbic acid of cabbage juice Effect of lactic acid fermentation on viable cell counts, LAB counts of cabbage juice Effect of lactic acid fermentation on coliform counts, yeast and mold of cabbage juice Effect of lactic acid fermentation on antimicrobial activity of cabbage juice Differentiation of quality of lactic acid bacteria of probiotic cabbage juices on the basis of physico-chemical parameters Differentiation of quality of lactic acid bacteria of probiotic cabbage juices on the basis of microbiological quality Effect of lactic acid fermentation on TSS, pH and titratable acidity of cucumber juice 66 4.16 4.17 4.18 4.19 4.20 4.21 4.22 4.23 4.24 4.25 4.26 4.27 4.28 4.29 4.30 4.31 4.32 4.33 4.34 4.35 4.36 68 69 69 70 73 75 76 78 80 81 81 82 85 87 88 89 92 93 94 94 97 Table 4.37 4.38 4.39 4.40 4.41 4.42 4.43 4.44 4.45 4.46 4.47 4.48 4.49 4.50 4.51 4.52 4.53 4.54 4.55 4.56 4.57 Title Effect of lactic acid fermentation on reducing sugar, total sugar and total phenols of cucumber juice Effect of lactic acid fermentation on antioxidant activity and ascorbic acid of cucumber juice Effect of lactic acid fermentation on viable cell counts and lactic acid bacteria counts of cucumber juice Effect of lactic acid fermentation on coliform counts ,yeast and mold of cucumber juice Effect of lactic acid fermentation on antimicrobial activity of cucumber juice Differentiation of quality of lactic acid bacteria of probiotic cucumber juices on the basis of physico-chemical parameters Differentiation of quality of lactic acid bacteria of probiotic cucumber juices on the basis of microbiological quality Effect of lactic acid fermentation on TSS, pH and titratable acidity of carrot juice Effect of lactic acid fermentation on total Sugar, reducing sugar and total phenols of carrot juice Effect of lactic acid fermentation on antioxidant activity and ascorbic acid during lactic acid fermentation of carrot juice Effect of lactic acid fermentation on viable cell counts and lactic acid bacteria counts of carrot juice Effect of lactic acid fermentation on coliform, yeast and mold counts of carrot juice Effect of lactic acid fermentation on antimicrobial activity of carrot juice Differentiation of quality of lactic acid bacteria of probiotic carrot juices on the basis of physico-chemical parameters Differentiation of quality of lactic acid bacteria of probiotic carrot juices on the basis of microbiological quality Evaluation of all probiotic vegetables (cabbage, cucumber and carrot) juices on the basis of rate of fermentation (°B/24hr) and rate of acidification (%/24hr) and lactic acid bacteria counts(108cfu/ml) Effect cold storage on physico-chemical parameters of apple probiotic juice Effect of cold storage on microbiological analysis of apple probiotic juice Effect cold storage on antimicrobial activity of apple probiotic juice Effect cold storage on antimicrobial activity of apple probiotic juice Effect of cold storage on physico-chemical parameters of pear probiotic juice Page 99 100 101 104 105 105 106 108 110 112 113 115 116 117 118 119 120 122 124 124 126 Table Title Page 4.58 Effect cold storage on microbiological analysis of pear probiotic juice Effect of cold storage on antimicrobial activity of pear probiotic juice Acid tolerance and bile tolerance of lactic acid bacteria of probiotic pear juice Effect of cold storage on physico-chemical parameters of orange probiotic juice Effect of cold storage on microbiological quality of orange probiotic juice Effect of cold storage on antimicrobial activity of orange probiotic juice Acid tolerance and bile tolerance of lactic acid bacteria of probiotic orange juice Effect of cold storage on physico-chemical parameters of cabbage probiotic juice Effect of cold storage on microbiological quality of cabbage probiotic juice Effect cold storage on antimicrobial activity (mm) of cabbage probiotic juice Acid tolerance and bile tolerance of lactic acid bacteria of probiotic cabbage juice Effect of cold storage on physico-chemical parameters of cucumber probiotic juice Effect of cold storage on microbiological quality of cucumber probiotic juice Effect cold storage on antimicrobial activity (mm) of cucumber probiotic juice Acid tolerance and bile tolerance of lactic acid bacteria of probiotic cucumber juice Effect of cold storage on physico-chemical parameters of carrot probiotic juice Effect of cold storage on microbiological quality of carrot probiotic juice Effect cold storage on antimicrobial activity of carrot probiotic juice Acid tolerance and bile tolerance of lactic acid bacteria of probiotic carrot juice Morphological, physiological and biochemical characteristics of lactic acid bacteria of probiotic fruit and vegetable juices Sensory characteristics of apple, pear and orange probiotic juices Sensory characteristics of cabbage, cucumber and carrot probiotic juice 128 4.59 4.60 4.61 4.62 4.63 4.64 4.65 4.66 4.67 4.68 4.69 4.70 4.71 4.72 4.73 4.74 4.75 4.76 4.77 4.78 4.79 129 130 131 133 135 135 137 139 140 141 142-143 144 145 145 148 150 151 152 154 155 156 LIST OF FIGURES Plate Title 2.1 2.2 2.3 3.1 3.2 3.3 3.4 3.5 3.6 4.1 Production growth trend for fruits and vegetables crops Lactic acid fermentation Metabolites of lactic acid bacteria Pictorial representation for preparation of probiotic apple Pictorial representation for preparation of probiotic pear Pictorial representation for preparation of probiotic orange Pictorial representation for preparation of probiotic cabbage Pictorial representation for preparation of probiotic cucumber Pictorial representation for preparation of probiotic carrot Effect of lactic acid fermentation rate of fermentation, rate of acidification, retention of total phenols and vitamin C of apple juice Changes in TSS (°B) , pH , titratable acidity (%) , total sugar (%), reducing sugar (%) and total phenols (mg/100g)(f) during lactic acid fermentation of apple juice Changes in antioxidant activity(%),vitamin C(mg/100g),lactic acid bacteria counts(cfu/ml),antimicrobial activity against S.aureus and E.coli during lactic acid fermentation of apple juice Effect of lactic acid fermentation rate of fermentation, rate of acidification, retention of total phenols and vitamin C of pear juice Changes in TSS (°B) , pH, Titratable acidity (%) , Total sugar (%) , Reducing sugar (%) and Total phenols (mg/100g)(f) during lactic acid fermentation of pear juice Changes in antioxidant activity(%),vitamin C(mg/100g),lactic acid bacteria counts(cfu/ml),antimicrobial activity against S.aureus and E.coli during lactic acid fermentation of pear juice Effect of lactic acid fermentation rate of fermentation, rate of acidification, retention of total phenols and vitamin C of orange juice Changes in TSS (°B) ,pH, Titratable acidity (%),Total sugar (%), Reducing sugar (%) and Total phenols (mg/100g)(f) during lactic acid fermentation of orange juice Changes in antioxidant activity(%),vitamin C(mg/100g),lactic acid bacteria counts(cfu/ml),antimicrobial activity(mm) against S.aureus and E.coli during lactic acid fermentation of orange juice 4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 Between Page(s) 12 15 30-31 30-31 30-31 30-31 30-31 30-31 44-45 46-47 54-55 58-59 58-59 64-65 66-67 72-73 76-77 ... is a bonafide research work carried out by Ms Anuradha Thakur (H-13-21-M) daughter of Shri Madan Singh under my supervision and that no part of this thesis has been submitted for any other degree... - II This is to certify that the thesis titled, “Development of probiotic juice by lactic acid fermentation of fruits and vegetables” submitted by Ms Anuradha Thakur (H-13-21-M) daughter of Shri... investigation and thesis preparation I earnestly acknowledge assiduous effort and sanguine gestures of Dr Anju Dhiman, Prof and Head Dept of Food Science and Technology, Mrs Surekha Attri, Dr KS Thakur,