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(Carica Papaya) PROTEASE in CREAM CHEESE PREPARATION by RESPONSE SURFACE METHODOLOGY

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(Carica Papaya) PROTEASE in CREAM CHEESE PREPARATION by RESPONSE SURFACE METHODOLOGY OPTIMIZATION OF CRUDE PAPAYA (Carica papaya) PROTEASE IN CREAM CHEESE PREPARATION BY RESPONSE SURFACE METHODOLOGY by Nabindra Kumar Shrestha Department of Food Technology Central Campus of Technology Institute of Science and Technology Tribhuvan University, Nepal 2019 Optimization of Crude Papaya (Carica papaya) Protease in Cream Cheese Preparation by Response Surface Methodology A dissertation submitted to the Department of Food Technology, Central Campus of technology, Tribhuvan University, in partial fulfilment of the requirements for the degree of B. Tech. in Food Technology by Nabindra Kumar Shrestha Department of Food Technology Central Campus of Technology Institute of Science and Technology Tribhuvan University, Nepal May, 2019 ii Tribhuvan University Institute of Science and Technology Department of Food Technology Central Campus of Technology, Dharan Approval Letter This dissertation entitled Optimization of Crude Papaya (Carica papaya) Protease in Cream Cheese Preparation by Response Surface Methodology presented by Nabindra Kumar Shrestha has been accepted as the partial fulfilment of the requirement for the B. Tech. degree in Food Technology. Dissertation Committee 1. Head of Department ___________________________________________ (Mr. Basanta Kumar Rai, Assoc. Prof.) 2. External Examiner ___________________________________________ (Mr. Shyam Kumar Mishra, Assoc. Prof.) 3. Supervisor ___________________________________________ (Mr. Bunty Maskey, Asst. Prof.) 4. Internal Examiner ___________________________________________ (Mr. Yadav K.C., Asst. Prof.) Co-supervisor ___________________________________________ (Mr. Ram Shovit Yadav, Asst. Prof.) May, 2019 iii Acknowledgements I would like to express my heartfelt gratitude to my respected guide Asst. Prof. Mr. Bunty Maskey for his kind support, excellent guidance, encouragement and constructive recommendations throughout the work. I am really thankful to Prof. Dr. Dhan Bahadur Karki (Campus Chief, Central Campus of Technology) and Assoc. Prof. Basanta Kumar Rai (Chairperson, Department of Food Technology) and Assoc. Prof. Geeta Bhattarai (Chairperson, Central Department of Food Technology) for providing opportunity and making available all the facilities to carry out the best in this dissertation work. I cannot stay from thanking all my classmates for their help in preparing me for this through all those experiences and memories which will never be forgotten in my whole life. Special thanks must be bestowed to my friends Anjana Shrestha, Bandana Panta, Bishesh Vishwokarma, Jitendra Shahi, Sandeep Lamichhane, Sanil Joshi, Sira Shrestha and all my beloved juniors for their support during the completion of this work. I would like to thank all those whose perceptions, observation and inputs have helped me directly or indirectly. I also express my sincere thanks to my teachers, staff of library and laboratory for their direct and indirect co-operation and suggestions without whom this work could not have been possible. No words in this world can ever express the love, encouragement, inspiration and moral support given by my parents and family members during my tough days. Date of submission: May, 2019 _____________________ (Nabindra Kumar Shrestha) iv Abstract The effect of papaya latex as crude papaya protease on physicochemical, microbial and sensorial characteristics of cream cheese prepared were examined. Enzyme concentration, pH of milk and temperature of milk on milk clotting activity were optimized by surface response methodology. Also, the protease activity of the crude enzyme was also measured using optimized conditions. Finally, the cheese prepared by using crude papaya protease was compared with that prepared using rennet and commercial papain for physicochemical, microbial and sensorial properties. Numerical optimization study revealed maximum milk clotting activity at pH 6.5, temperature 70℃ and enzyme concentration 1% (1 ml/1000 ml milk) using papaya protease as coagulant. Protein, ash and calcium showed no significant (p>0.05) difference among the cheeses made using different coagulants. However, significantly (p<0.05) higher levels of moisture and ash, and lower levels of fat were observed in the cheese produced by papaya protease compared to that made using rennet. Papaya protease significantly enhanced the spreadability of cheese while the other sensory properties were similar to the control except aftertaste. Microbiologically, all samples were not significantly different and were free of coliforms. However, the presence of TPC and yeasts and molds was found. The results revealed that the papaya latex as crude papaya protease may have potential application for the manufacture of cream cheese and further could be utilized as a milk coagulant in cheese making. v Contents Approval Letter .................................................................................................................. iii Acknowledgements ............................................................................................................. iv Abstract ................................................................................................................................ v List of Tables .............................................................................................................. xiii-xiv List of Figures .................................................................................................................... xv List of Abbreviations ................................................................................................ xvi-xvii 1. Introduction .................................................................................................................. 1-4 1.1 General introduction .................................................................................................... 1 1.2 Statement of the problem ............................................................................................. 2 1.3 Objectives .................................................................................................................... 3 1.3.1 General objective ................................................................................................ 3 1.3.2 Specific objectives .............................................................................................. 3 1.4 Significance of the study .............................................................................................. 3 1.5 Limitations of the study ............................................................................................... 4 2. Literature review ........................................................................................................ 5-46 2.1 History and development of cheese ............................................................................. 5 2.1.1 Scientific and technological developments ......................................................... 7 2.2 Cheese production ........................................................................................................ 8 2.2.1 Cheese production- a Nepalese scenario ............................................................. 8 2.3 Varieties and classification .......................................................................................... 9 2.4 Cream cheese ............................................................................................................. 13 vi 2.4.1 History of cream cheese .................................................................................... 14 2.4.2 Major brands of cream cheese .......................................................................... 14 2.4.3 Uses of cream cheese ........................................................................................ 14 2.5 Quality of milk in relation to cheese making ............................................................. 15 2.5.1 The amount and composition of milk constituents ........................................... 15 2.5.1.1 Milk protein (casein) and its effect ....................................................... 16 2.5.1.2 Milk fat ................................................................................................. 16 2.5.1.3 Milk salt ............................................................................................. 17 2.5.2 Abnormal milk .................................................................................................. 18 2.5.2.1 Mastitis milk ......................................................................................... 18 2.5.2.2 Colostrum ........................................................................................... 19 2.5.2.3 Late lactation milk ............................................................................. 19 2.5.3 Changes in the milk after production ................................................................ 19 2.5.3.1 Changes in casein micelles and salt equilibrium ............................... 19 2.5.4 Inhibitory substances ......................................................................................... 20 2.5.4.1 Natural inhibitory systems in milk ....................................................... 20 2.5.4.2 Antibiotic residues ................................................................................ 20 2.5.4.3 Residues of detergents and sanitizers ................................................ 21 2.5.5 Microbiological quality of milk .......................................................................
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