Variation of gaba content and physical properties of sprouted mung bean fortified yoghurt

THAI NGUYEN UNIVERSITY OF AGRICULTURAL AND FORESTRY KIMBERLY ANNE A BAUTISTA VARIATION IN GAMMA AMINOBUTYRIC ACID, PHYTIC ACID CONTENT AND PHYSICAL PROPERTIES OF GERMINATED MUNG BEAN-FORTIFIED YOGHURT BACHELOR THESIS Study Mode: Major : Faculty : Batch : Full-time Food Technology Advanced Education Program Office 2017-2021 Thai Nguyen, 27/05/2022 DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Food Technology Student name Kimberly Anne A Bautista Student ID DTN1754190022 Thesis Title Variation of GABA Content and Physical Properties of Sprouted Mung-Bean Fortified Yoghurt Supervisor (s) Associate Professor Dr Nguyen Duc Doan and Dr Condro Wibowo Supervisor’s Signature Abstract: Wibowo Yoghurt and mung beans each have their benefits in the aspect of health The effect of germinated mung beans and storage time on the properties of yoghurt are determined Germinated mung bean-fortified yoghurt having high gammaaminobutyric acid (GABA) and low phytic acid (PA) content with competent physical and sensory qualities is produced in aiding the need to improve health This project followed certain analytical methods for the determination of total dry matter content, protein content, GABA content, PA content, acidity, pH level, and water holding capacity 2-factor ANOVA analysis and Tukey comparison were used for data processing The results concluded that the protein and GABA content rises with the temperature On the contrary, PA and total dry matter content lower when the germination time and temperature increase Further results state that storage time modifies the viscosity, acidity, pH, water holding capacity, protein, PA, GABA, and total dry matter of yoghurt However, it does not exceptionally affect the physical ii properties of the yoghurt The project then provides recommendations for future researches regarding this topic It is suggested to expand the germination temperature range to more specifically monitor the change of components in mung bean seeds and, to study some other lactic acid bacteria that are capable of synthesizing GABA in yoghurt Keywords: Number of pages: mung bean, germination, yoghurt, GABA, PA 53 Date of Submission: May 2022 iii Acknowledgement I would like to state my sincere gratitude and respect to all those who contributed and gave guidance for the duration of this thesis The success of this research paper would not be possible without the knowledge, guidance, and support of my supervisor A/Prof Dr Nguyen Duc Doan I would also like to give thanks to Dr Condro Wibowo who co-supervises the entirety of this paper My gratitude also goes to the Advance Education Program of Thai Nguyen University of Agriculture and Forestry for the support they have given to all of the students in their final year Great thanks to Vietnam National University of Agriculture for allowing me to use their laboratory and equipment which are necessary for the completion of this dissertation I would also like to thank Nguyen Thi Thu Thuy for helping with the usage of the laboratory and its equipment The next ones I would like to thank are my friends who gave simple advice and discussions during mental blockage I would also like to gratify my family for their continuous encouragement Lastly and most importantly, I would like to state my deepest gratitude to God who made everything in life a possibility iv TABLE OF CONTENT List of Figures List of Tables List of Abbreviations PART I INTRODUCTION 1.1 Research rationale 1.2 Research’s objectives 1.3 Research questions and hypotheses 1.4 Limitations PART II LITERATURE REVIEW PART III METHODS 11 3.1 Material and chemicals 11 3.2 Methods 12 3.2.1 Production process of germinated mung bean yoghurt 12 3.2.2 Analytical method 13 3.2.2.1 Determination of total dry matter content 13 3.2.2.2 Determination of protein content 14 3.2.2.3 Determination of GABA content 15 3.2.2.4 Determination of phytic acid content 16 3.2.2.5 Determination of the PH of yoghurt 17 v 3.2.2.6 Determination of the acidity of yoghurt 17 3.2.2.7 Determination of the viscosity of yoghurt 18 3.2.2.8 Determination of water holding capacity (WHC) 18 3.2.2.9 Sensory assessment 19 3.2.3 Data processing method 19 PART IV RESULTS 21 4.1 Chemical composition of raw green beans 21 4.2 Total dry matter content in germinated mung beans 21 4.3 Content of gamma-aminobutyric acid in mung bean seeds germination 22 4.4 Phytic acid content in germinated mung beans 22 4.5 Protein content in germinated green beans 23 4.6 Total dry matter content of germinated mung bean yoghurt during cold storage ……………………………………………………………………………………23 4.7 Gamma-aminobutyric acid content of germinated mung bean yoghurt during cold storage 24 4.8 Phytic acid content of germinated mung bean yoghurt during cold storage 24 4.9 Protein content of germinated mung bean yoghurt during cold storage 25 4.10 Effect of storage time on pH of germinated green bean yoghurt 25 4.11 Effect of storage time on acidity of mung bean yoghurt germination 26 4.12 Effect of storage time on the viscosity of mung bean yoghurt germination………………………………………………………………… … 26 vi 4.13 Effect of storage time on water holding capacity of mung bean yoghurt germination 27 4.14 Sensory quality of mung bean yoghurt 27 PART V DISCUSSION AND CONCLUSION 29 5.1 Discussion 29 5.2 Conclusion 35 REFERENCES 37 APPENDICES 42 vii LIST OF FIGURES Figure Effect of time and temperature on total dry matter content of mung beans ……………….………………………………………………………… ……… 21 Figure Effect of time and temperature on GABA content of mung beans …… 21 Figure Effect of time and temperature on PA content of mung beans……… …22 Figure Effect of time and temperature on protein content of mung beans.…… 22 Figure Effect of cold storage time on total dry matter content of mung bean yoghurt …………………………………………………………………… … 23 Figure Effect of cold storage time on GABA content of mung bean yoghurt… 23 Figure Effect of cold storage time on PA content of mung bean yoghurt…… 24 Figure Effect of cold storage time on protein content of mung bean yoghurt… 24 Figure Effect of cold storage time on the pH level of mung bean yoghurt…… 25 Figure 10 Effect of cold storage time on the acidity of mung bean yoghurt…… 25 Figure 11 Effect of cold storage time on the viscosity of mung bean yoghurt……26 Figure 12 Effect of cold storage time on WHC of mung bean yoghurt……… …26 Figure 13 Mung bean yoghurt final product……………………………… …….26 Figure 14 Sensory evaluation results of mung bean yoghurt…………… ………27 LIST OF TABLES Table The 9-Point Hedonic Scale .19 Table Chemical parameters of raw green peas 20 Table Statistical processing table of effects of temperature and germination time on GABA, PA and chemical composition of mung bean seeds .20 LIST OF ABBREVIATIONS FeCl3 iron (III) chloride GABA gamma-aminobutyric Acid H2NaO+ sodium hydrate H2 O water H2SO4 sulfuric acid HClO4 perchloric acid NaOH sodium hydroxide NH3 ammonia PA phytic acid TUAF Thai Nguyen University of Agriculture and Forestry VNUA Vietnam National University of Agriculture WHC water holding capacity magnesium salts form which may be the reason for the decrease in PA content (Cheryan & Rackis, 1980), this is removed along with the bean residue through the pouring filter Furthermore, Bilgicli & Isbanotlu (2007) demonstrated that PA content reduces by more than 90% during lactic fermentation Therefore, the decrease in PA content is possibly due to the activity of the phytase enzyme in the fermented milk mixture secreted by lactic acid bacteria The protein content of yoghurt is significantly affected by the storage time (P < 0.001) Contrary to the increase of dry matter content, the protein content of yoghurttends to decrease when the storage time approaches 21 days In a more detailed sense, the control had the lowest content of protein and gradually increases at day 1and reaching its peak at day then declining at day 14 and 21 The protein content decreases during the cold storage process since proteolysis occurs to form amino acids and peptide chains (Kiel, 2012) The amino acids are partially released to create the aroma of the finished yoghurt The increase in protein content of fermented yoghurt compared to pre-fermentation mixed milk solution is due to the occurrence of protein biosynthesis As shown in figure 9, we can see that the pH of yoghurt decreases steadily with storage time During the preservation of yoghurt at a temperature of 2-6℃, the fermentation process still takes place to form lactic acid which increases the acidity and decreases the pH of the yoghurt The pH level diminished intensely during the storage period from 14 to 21 days In conclusion, there is an inverse relationship between the pH level of yoghurt and storage time When the storage time gets longer, the pH of the yoghurt decreases 33 Figure 11 shows that the viscosity of the yoghurt sample increased progressively with the storage time Yoghurt stored after days and had a viscosity range of 8.32 - 8.40 P.s then increased slightly on days 14 and 21 from 8.62 to 8.85 Pa.s reaching its pinnacle with the maximum amount of bacteria Research results show that the viscosity of yoghurt increases slightly with prolonged cold storage According to Saint-Eve et al (2008), acidification during yoghurt storage complexly explained the increase in viscosity The water holding capacity of yoghurt tends to increase with cold storage time The results obtained showed that the most significant change was at day to day 14 The increased water-holding capacity of yoghurt during storage was because of the continued occurrence of fermentation during cold storage which causes the dry matter content to slightly increase The stated phenomenon leads to the retention of water Research results of Erfanian & Rasti (2019) have demonstrated that the structure of yoghurt depends on the dry matter content In parallel with the obtainedresults and the fact that the product has a poorer water retention capacity, added water won't mix which leads to the decrease of quality of the finished yoghurt Figure 14 shows that the color index of yoghurt had the highest score (5.70), followed by the indicator of texture (5.63) The remaining parameters: smell and taste had no significant difference However, the actual results obtained show that there is still a noticeable difference in the odor and taste criteria It can be noted thatthe germinated mung bean yoghurt product has an unfavorable smell from the beanswhich somehow contributes to the taste In summary, when it comes to the 34 sensory properties of the product which is the mung bean yoghurt, odor and taste indicators is much lower than the color and texture criteria 5.2 Conclusion This study shows the importance of germination time, temperature, and the interaction between both to GABA, PA, and composition of mung beans Elements were affected significantly by the temperature, time, and the interaction between the two However, GABA was not significantly affected by time while PA was not significantly affected by the interaction between the two-factors Under research conditions, total dry matter content decreased gradually with increasing time and temperature, obtaining the highest percentage in 12 hours with the temperature of 28°C Germinated mung beans showed a higher GABA content and with the experiment, it is known that GABA content rises along with the temperature having the highest content in 12 hours at 30°C In terms of PA content, it drops as the germination time increases at all temperatures The minimum amount of PA was noted with the germination time of 24 hours In addition, as the time of germination extends in all temperature, the protein content increases, obtaining the highest percentage at 32°C after 24 hours Storage time influences viscosity, acidity, pH, protein content, PA, WHC, GABA, and total dry matter of yoghurt However, it does not remarkably affect the physical properties of yoghurt 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sữa chua đậu nành nảy mầm TCVN (4295:2009) Tiêu chuẩn Quốc giavề Đậu hạt – Phương phápt TCVN 6509:201 (ISO 11869:2012) Sữal ên men – Xác định độ axit chuẩn – Phương pháp đo điện 41 TCVN 8125:2009 (ISO 20483:2006) Tiêu chuẩn Quốc giavề Ngũ cốc, đậu đỗ Xác định hàm lượng ni tơ tính hàm lượng protein thơ – phương pháp Kjeldahl Trương Nhật Trung & Đống Thị Anh Đào (2016) Làm giàu hàm lượng gammaaminobutyric acid (GABA) hạt đậu xanh điều kiện nảy mầm hypoxiaanaerobic đánh giá hao tổn sau trình luộc Tạp chí Khoa học Phát triển Cơng nghệ 19(K7): 88-96 42 APPENDICES Appendix 1: Summary of the effects of temperature and germination time on GABA, PA and other chemical compositions of mung beans Ungerminated Protein Total % dry matter matter mg/g % matter 7.72 ± 0.50 mung bean 40.94 dry GABA PA dry mg/g dry matter ± 3.29 ± 0.40 19.60± 0.39 ± 3.24b± 0.13 15.35b 0.26 Temperature °C 28 8.19b ± 0.10 41.60a 0.47 30 9.15a ± 0.30 35.89c 0.69 ± 3.54a ± 0.22 0.70 32 8.90a ± 0.23 37.53b ± 16.28a ± 0.46 ± 2.89c ± 0.17 0.31 15.54ab ± 0.56 Time (hour) 12 8.35b ± 0.22 40.60a ± 3.21a ± 0.12 0.39 18 8.58b ± 0.23 38.91b 16.50a ± 0.25 ± 3.28a± 0.22 15.91a ± 0.71 ± 3.18a ± 0.18 14.77b ± 0.74 0.74 24 9.31a ± 0.17 35.51c 0.65 Anova 43 Temperature *** *** *** *** Time *** *** n/s *** * *** *** *** n/s Temperature time Note: The data with letters a, b, c different by column under the same germination conditions are significantly different: *** P < 0.001; n/s: P > 0.05 Appendix 2: Summary table of effects of storage time on GABA, PA and other chemical compositions of mung bean yoghurt Protein Total dry GABA PA % dry matter matter mg/g % matter matter dry mg/g dry Control 11.47c ± 0.47 14.17c ± 0.20 3.81a ± 0.62 32.07a ± 0.61 Day 15.09ab ± 0.87 15.48b ± 0.12 1.58b ± 0.32 25.77b ± 0.61 Day 15.75a ± 0.45 15.60b ± 0.26 1.56b ± 0.26 24.40c ± 0.21 Day 14 14.89ab ± 0.86 16.20a ± 0.21 1.33b ± 0.31 24.29c ± 0.30 Day 21 13.05b ± 0.67 1.22b ± 0.19 23.54c ± 0.16 16.29a ± 0.63 Note: Data with letters a,b,c represent the difference between different storage days (P