MINISTRY OF EDUCATION AND TRAINING HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY FOR HIGH QUALITY TRAINING GRADUATION PROJECT CHEMICAL AND FOOD TECHNOLOGY ACQUISITION OF PROTEIN FROM LIMA BEAN (PHASEOLUS LUNATUS) WITH HIGH RECOVERY EFFICIENCY LECTURER: PhD. PHAM THI HOAN STUDENT: LE DUC HUY HOANG TRAN HUY SKL012527 Ho Chi Minh City, January 2024 HCMC UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY OF CHEMICAL AND FOOD TECHNOLOGY FOOD TECHNOLOGY GRADUATION THESIS CODE: 2023-19116041 ACQUISITION OF PROTEIN FROM LIMA BEAN (PHASEOLUS LUNATUS) WITH HIGH RECOVERY EFFICIENCY INSTRUCTOR: PhD. PHAM THI HOAN STUDENTS: LE DUC HUY HOANG 19116041 TRAN HUY 19116034 HO CHI MINH CITY – 01/2024 THE SOCIALIST REPUBLIC OF VIETNAM Independence – Freedom– Happiness -------- GRADUATION THESIS ASSIGNMENT Student name: Le Duc Huy Hoang Student ID: 19116041 Student name: Tran Huy Student ID: 19116034 Major: Food Techonology Class: 19116CLA Supervisor: Pham Thi Hoan, Ph. Email: hoanpt@hcmute.vn Date of assignment: 21/08/2023 Date of submission: 22/01/2024 1.
Thesis title: Acquisition of protein from Lima bean (Phaseolus lunatus) with high recovery efficiency. Thesis assignment: - Synthesize scientific documents and articles, combine and experiment to select an appropriate technological process in acquisition of protein from Lima bean with high recovery efficiency. - Conduct surveys and evaluate some product properties such as chemical compositions, structure and molecular weight of proteins obtained from Lima protein. - Proposing directions for applying protein from Lima bean in the field of food technology.
The content and requirements of the graduation thesis have been approved by the Chair of the Food Technology program Ho Chi Minh City, January 22th , 2024 CHAIR OF THE PROGRAM SUPERVISOR (Sign with full name) (Sign with full name) ACKNOWLEDGEMENTS First of all, allow us to express our most respection to all the teachers are working at Ho Chi Minh City University of Technology and Education in general, and to all the Asscociate Professors, Doctors of Philosophy and Masters at the Faculty of Chemical and Food Technology in particular. Throughout the process of learning and growing up at university, undergo a thorough and complete training program, along with the enthusiastic teaching and support of teachers, we have been equipped with knowledge from basic to advanced, from general to academic in the field we are pursuiting. The studies acquired from the university environment along with the practical sharing from the experiences of the teachers will certainly be valuable tools on the upcoming working path in larger and realistic atmospheres. Most especially, we would like to express our deep appreciation to Doctor.
Pham Thi Hoan, a lecturer working at the Faculty of Chemical and Food Technology, who has accepted and instructed us to complete this graduation thesis. In addition to provide support and direction for the thesis, we also received a lot of support from the instructor in terms of chemicals, machines and equipments needed to complete the experiments. Besides, we would also like to send an appreciation to Mrs. Ho Thi Thu Trang - faculty laboratory manager.
Respect to her dedicated help with tools, laboratory rooms and instructions for operating of various equipments, we were able to complete our graduation thesis. Finally, we would like to show our love to fellow classmates, who worked on the graduation thesis together, along with family and friends, for their support and encouragement throughout the thesis process in the laboratory. Due to our lack of experiences, knowledge, and also limited time, there might be some mistakes during processing the thesis. We are looking forward to be forgiven and always get ready to hear your contributions to assist us in improving our topic.
Finally, we would like to send our best wishes to the Faculty of Chemical and Food Technology. Wishing health and success to everyone. Sincerely thank, Le Duc Huy Hoang and Tran Huy. i DECLARATION We declare that the Graduation Thesis “Acquisition of protein from Lima bean (Phaseolus lunatus) with high recovery efficiency” is the result of our study and has never been published in other studies before.
We took research ethics seriously during the implementation process of the thesis. All the results of this project are the result of our researches and experiences, and all references in this project are properly referenced in accordance with rules. We are solely responsible for the accuracy of the numbers, statistics, and other aspects of our graduation thesis. Ho Chi Minh city, January.
,2024 (Sign with full name) ii TABLE OF CONTENTS ACKNOWLEDGEMENTS. ii ASSESSMENT FORM GRADUATION THESIS OF FOOD TECHNOLOGY .iii ASSESSMENT FORM GRADUATION THESIS OF FOOD TECHNOLOGY. iv ASSESSMENT FORM GRADUATION THESIS OF FOOD TECHNOLOGY. v TABLE OF CONTENTS.
vi LIST OF FIGURES. ix LIST OF TABLES. x LIST OF ABBREVIATIONS. Research object and Scope of research.
Scope of the research. Scientific and practical significance. Overview of material. Introduction to legume.
Introduction to Lima bean. Classification of Lima bean. Nutritions of Lima bean.2 Overview of protein. Introduction to protein.
Introduction to legume protein. Introduction to Lima protein. Recovery methods of protein. Isoelectric point precipitation.
Organic solvent cause precipitation. Research situations of the topics. MATERIALS AND METHODS. Materials, chemicals, devices and equipments.
Equipment and Devices. Research framework and protein collection process. Diagram of protein collection. Determine the extraction yield of protein.
Determine the protein recovery yield of protein. Determine the physicochemical properties of protein. Determine the functional properties of protein. RESULTS AND DISCUSSIONS.
Determine the appropriate extraction regime. Determine the physicochemical properties of protein. Determination of chemical compositions of Lima powder and Lima protein. Determination of secondary structure of protein (FTIR).
Determination of molecular weight of protein (SDS-PAGE). Determine the functional properties of protein. Determine the water/oil absorption capacity (WAC - OAC). Determine the foam capacity and stability (FC – FS).
Determine the emulsifying activity index and stability (EAI – ESI). Determine the solubility. CONCLUSIONS AND RECOMMENDATIONS. 68 viii LIST OF FIGURES Figure 2.
Production of grain legumes in the 12 European countries where the total harvest exceeded 100,000 t in 2013. Data from Eurostat (2016) (faba bean, pea, soybean, lupins) and FAOSTAT (2016) (lentil, chickpea, common bean). Baby Lima beans. Fordhook Lima beans.
Speckled Lima bean. Diagram of protein collection. Scheme of experimental design 1. Scheme of experimental design 2.
Complete research diagram of protein collection. Results of FTIR from Lima protein (PCS10) and Soy protein (SPI). Results of SDS-PAGE from Lima and Soy protein sample. Calibration curve between logarithm of molecular weight and travel distance of proteins.
Influence of pH of PCS10 suspension on foam capacity. Influence of pH of SPI suspension on foam capacity. Result of EAI of Lima protein (PCS10) and Soy protein (SPI). Result of ESI of Lima protein (PCS10) and Soy protein (SPI).
Result of the solubility of Lima protein. 55 ix LIST OF TABLES Table 2. Description of commercially available Lima bean varieties. Proximate composition of Lima bean samples (%).
The mineral content of the Lima bean samples (mg/100g). The amounts of amino acids in Lima bean seeds. Results of experimental design 1. Results of experimental design 2.
Chemical compositions of Lima powder and Lima protein. Results of WAC and OAC from proteins. 49 x LIST OF ABBREVIATIONS SPI: Soy Protein Isolate WHC: Water Holding Capacity OHC: Oil Holding Capacity FC: Foam Capacity FS: Foam Stability EAI: Emulsion Ability Index ESI: Emulsion Stability Index RPM: Round per minute SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis FTIR: Fourier-transform infrared spectroscopy xi ABSTRACT Currently, the demand for using plant-based protein sources to partially or entirely replace animal protein in human daily meals is increasing. Lima bean (Phaseolus lunatus) is a rich source of plant-based protein and abundant in nutrients but their researches in protein are not fully exploited, especially in Viet Nam.
This research was conducted to extract protein from Lima bean, determine physicochemical properties (chemical compositions, molecular weight and secondary structure of the protein), as well as the functional properties (oil absorption, water absorption, foaming ability, emulsification capacity and solubility) of the extracted protein. The research results showed that the protein acquisition process, using a combination of solvent precipitation and isoelectric point precipitation at pH 4.5, tempereture of 4°C, achieved recovery efficiency at 82.14% higher than that in the other samples. The collected sample includes the moisture, protein, fat, ash, and total carbohydrate content of 10. The molecular weight and secondary structure of Lima bean protein also reflected similarities with commercially available soy protein isolate (SPI) on the market.
Lima bean protein exhibited water absorption capacity nearly similar to the control sample (soy protein isolate - SPI), with values of 3. Foaming ability and foam stability were also similar to soy protein isolate, reaching the highest values at pH 2 and 10; emulsification capacity and emulsion stability were highest at pH 10 and lowest at pH 4; solubility of Lima bean protein was higher than SPI at pH levels outside the isoelectric point of the protein (pH 2, 6, 8, and 10) and reaching the highest value at pH 10. The results of this study indicate that Lima bean protein can be applied in various manufacturing fields such as meat processing technology, ice cream production or bakery and confectionery production, etc. The effectiveness of Lima bean protein in specific applications will be further clarified in our research.
Problem statement The currently trend in human dietary habit involves the combination or partial replacement of plant-derived protein with animal-derived protein (Alves & Tavares 2019). This approach is gradually becoming popularity because there are some evidences can prove that a balanced diet incorporating both protein sources can contribute to lowering the risk of common human ailments, including cardiovascular disease, type 2 diabetes, and issues related to overweight and obesity (Ferrari et al. Throughout history and up to the present day, the nutritional contributions from plants have consistently held a significant role in the daily diets of human beings. Around the world, legumes play an important part in humanity diet.
Legume is considered the second most important food source for humans in a variety of locations of the world, after cereals, which are an essential part of the human diet. Especially in developing countries , they serve as an affordable source of protein when animal protein is lacking. Similar to grains and oil seeds, legumes are a great source of protein (22. Fiber, vitamins, minerals and complex carbohydrates can all be found plentiful in legumes, which play an important role in human diet, contribute to the health of body (Du et al.
Moreover, the food industry makes extensive use of some species of legumes, such soybean and common beans, because of their numerous functional qualities (like viscosity, emulsion and gelling abilities or foaming), which provide texture and sensory qualities (David Betancur-Ancona et al. Legumes also have a high lysine level, which makes them ideal for intergrating with other foods. The nutrient-dense seeds have an important role in biodiversity, sustainable agriculture, food security, and minimizing environmental change (da Silva et al. Lima bean (Phaseolus lunatus L) are a noteworthy member of the Legume family, Fabaceae, a high protein content species range from 21-26% (M.
Sandoval-Peraza et al., 2020), also known as “butter bean” in some regions, are renowned for their nutritional value and versatility in culinary applications. The Lima bean seeds grow from specific areas will have distinct characteristics from others. Originating from South America, particularly the Andes region, Lima beans have become a globally recognized legume. Nowadays, there are two popular types of protein: Protein Concentrate (PC) and Protein Isolate (PI) are available under powder form.
The way of deriving them are from alkaline- 1 based protein extraction processes.