IMPACT OF SELECTED YEASTS AND GLYCOSIDASE ON CHEMICAL COMPOSITION OF MANGO WINE LI XIAO NATIONAL UNIVERSITY OF SINGAPORE 2013 IMPACT OF SELECTED YEASTS AND GLYCOSIDASE ON CHEMICAL COMPOSITION OF MANGO WINE LI XIAO (B. Sc. (Hons.), NTU) A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF CHEMISTRY NATIONAL UNIVERSITY OF SINGAPORE 2013 THESIS DECLARATION I hereby declare that this thesis is my original work and it has been written by me in its entirety, under the supervision of Dr Liu Shao Quan, (in the Food Science and Technology research laboratory, S13-05), Chemistry Department, National University of Singapore, between July 2009 and August 2013. I have duly acknowledged all the sources of information which have been used in the thesis. This thesis has not been submitted for any degree in any university previously. The content of the thesis has been partly published in: 1. Li, X., Yu, B., Curran, P., & Liu, S. Q. (2011). Chemical and volatile composition of mango wines fermented with different Saccharomyces cerevisiae yeast strains. South African Journal of Enology and Viticulture, 32, 117-128. 2. Li, X., Yu, B., Curran, P., & Liu, S. Q. (2012). Impact of two Williopsis yeast strains on the volatile composition of mango wine. International Journal of Food Science and Technology, 47, 808-815. 3. Li, X., Chan, L. J., Yu, B., Curran, P., & Liu, S. Q. (2012). Fermentation of three varieties of mango juices with a mixture of Saccharomyces cerevisiae and Williopsis saturnus var. mrakii. International Journal of Food Microbiology, 158, 28-35. 4. Lee, P. R., Li, X., Yu, B., Curran, P., & Liu, S. Q. (2012). NonSaccharomyces yeasts and wine. In A. S. Peeters (Ed.), Wine: Types, Production and Health, (pp. 319-333). Hauppauge, New York, USA: Nova Science Publishers. 5. Chan, L. J., Lee, P. R., Li, X., Chen, D., Liu, S. Q., & Trinh, T. T. T. (2012). Tropical fruit wine: an untapped opportunity. In D. Cabel (Ed.), Food and Beverage Asia. Dec/Jan 2011/2012 (pp. 48–51). Singapore: Pablo Publishing Pte Ltd. ISSN: 2010-2364. i 6. Li, X., Chan, L. J., Yu, B., Curran, P., & Liu, S. Q. (2013). Influence of Saccharomyces cerevisiae and Williopsis saturnus var. mrakii on mango wine characteristics. Acta Alimentaria, Accepted (in press). 7. Li, X., Lim, S. L., Yu, B., Curran, P., & Liu, S. Q. (2013). Impact of pulp on the chemical profile of mango wine. South African Journal of Enology and Viticulture, 34, 119-128. 8. Li, X., Lim, S. L., Yu, B., Curran, P., & Liu, S. Q. (2013). Mango wine aroma enhancement by pulp contact and β-glucosidase. International Journal of Food Science and Technology, 48, 2258-2266. Li Xiao Name Aug 2013 Signature ii Date ACKNOWLEDGEMENTS I would like to express the deepest appreciation to my supervisor Dr Liu Shao Quan for his valuable advice, supervision, guidance and encouragements throughout the entire project. I feel honored to work with him and I have learned extensively from his creative ideas, profound knowledge, and in-depth research experience in the four years. I am also deeply grateful for his kindness and patience throughout the four years and his countless efforts in guiding me to complete my research project. Without his guidance and unwavering help this thesis would not have been possible. Next, I give my warmest appreciation to Dr Yu Bin who guided me extensively on techniques of SPME-GC-FID/MS and relative method development. I also thank the flavorists from Firmenich Asia for their generous sharing of their knowledge and experience in flavor science and helpful support on the wine sensory tests. I would also thank other professors of Food Science and Technology (FST) program, such as Prof. Zhou Wei Biao, Dr Huang Dejian, Dr Yuk Hyun Gyun and Dr David Popovich, for their teaching and enlightenment to expand my knowledge in different aspects of food science through my four-year study. In addition, a thank you to my previous honors year students Mr. Lim Sien Long and Miss Chew Xue Li for their help and commitment in conducting several experiments. I would also thank my fellow postgraduate students, Dr Lee Pin Rou, Ms Sun Jing Can, Ms Cheong Mun Wai, Ms Chen Dai and Ms Chan Li Jie for their suggestions in experiments and moral encouragement. I am also grateful for all the technical support provided by FST staff Ms Lee Chooi Lan, Ms Lew Huey Lee, Ms Jiang Xiaohui, Mr. Abdul Rahaman and Ms Maria Chong. Last but not least, I would like to thank my parents and my adorable fiancée Tan Rui for their persistent love, support and encouragement in my life. iii TABLE OF CONTENTS THESIS DECLARATION i ACKNOWLEDGEMENTS iii SUMMARY ix LIST OF TABLES xi LIST OF FIGURES xiii LIST OF ABBREVIATIONS xvii LIST OF SYMBOLS xix CHAPTER Introduction and Literature Review 1.1 Mango 1.2 Mango wine .2 1.3 General comments of wine flavor .5 1.4 Flavor modulation by different techniques .8 1.4.1 Selection of Saccharomyces .9 1.4.2 Selection of non-Saccharomyces 10 1.4.3 Mixed-culture fermentation 11 1.4.4 Yeast interactions in mixed-culture fermentation 13 1.4.5 Enzymes .15 1.5 Objectives of project .16 CHAPTER Materials and Methods .18 2.1 Fruits and yeast strains 18 2.2 Preparation of mango juice and pre-culture 19 2.3 Fermentation 20 2.4 Instrumental and sensory analysis of wine samples 23 CHAPTER Chemical composition of mango wines fermented with different Saccharomyces cerevisiae yeasts strains 27 iv 3.1 Introduction .27 3.2 Result and discussion 27 3.2.1 Brix, pH and yeast growth 27 3.2.2 Changes of sugars and organic acids 28 3.2.3 Volatile compounds in fresh mango juice 29 3.2.4 Volatile composition of mango wines after 14-day fermentation and kinetic changes of major volatiles .30 3.2.4.1 Alcohols 38 3.2.4.2 Esters .40 3.2.4.3 Volatile fatty acid 42 3.2.4.4 Carbonyl compounds 43 3.3 Conclusion .44 CHAPTER Impact of two Williopsis yeast strains on the volatile composition of mango wine 45 4.1 Introduction .45 4.2 Results and discussion .45 4.2.1 Yeast growth, sugar consumption, and acidity .46 4.2.2 Volatile compounds and their evolution change 47 4.2.3 Quantification of major volatile compounds 52 4.3 Conclusion .55 CHAPTER Influence of mixed-starter of Saccharomyces cerevisiae and Williopsis saturnus var. mrakii on mango wine characteristics 56 5.1 Introduction .56 5.2 Results and discussion .56 5.2.1 Yeast ratio screening and population evolution .57 5.2.2 Brix, pH, sugars, ethanol, glycerol and organic acids 58 v 5.2.3 Evolution of volatiles throughout the fermentation .59 5.2.3.1 Terpenes 59 5.2.3.2 Alcohols 61 5.2.3.3 Fatty acids .64 5.2.3.4 Esters .64 5.2.3.5 Other minor volatiles 66 5.2.4 Sensory test .67 5.3 Conclusion .68 CHAPTER Fermentation of three varieties of mango juices with a mixture of Saccharomyces cerevisiae and Williopsis saturnus var. mrakii .69 6.1 Introduction .69 6.2 Results and discussion .69 6.2.1 Changes in yeast population, pH, oBrix, sugars and organic acids 69 6.2.2 Volatiles composition and their kinetic changes in mango juices and wines .73 6.2.2.1 Terpenes 73 6.2.2.2 Alcohols 80 6.2.2.3 Esters .81 6.2.2.4 Carbonyls 81 6.2.2.5 Volatile fatty acids 82 6.2.3 Sensory test .82 6.3 Conclusion .83 CHAPTER Antagonistic mechanism between Saccharomyces cerevisiae and Williopsis mrakii in co- and sequential cultures .85 7.1 Introduction .85 7.2 Results and discussion .85 vi 7.2.1 Early death of W. mrkaii in co-culture with S. cerevisiae 85 7.2.2 Effect of cell density on growth of two yeasts .87 7.2.3 Death of S. cerevisiae in sequential inoculation with W. mrakii 90 7.2.4 Presence of toxic compounds in W. mrakii culture 90 7.3 Conclusion .94 CHAPTER Impact of pulp contact on development of chemical profile of mango wine .95 8.1 Introduction .95 8.2 Results and discussion .95 8.2.1 Yeast growth, oBrix and pH changes 95 8.2.2 Sugar and organic acid contents before and after fermentation .98 8.2.3 Ethanol and glycerol contents of mango wine .98 8.2.4 Volatile profile of mango juice and wine .99 8.2.4.1 Major volatiles in juice .99 8.2.4.2 Evolution of terpenes hydrocarbons .99 8.2.4.3 Evolution of alcohols 105 8.2.4.4 Evolution of fatty acids .106 8.2.4.5 Evolution of esters 107 8.2.4.6 Evolution of sulfur compounds .108 8.3 Conclusion .109 CHAPTER Enhancement of mango wine aroma by β-glucosidase 110 9.1 Introduction .110 9.2 Results and discussion .110 9.2.1 Physicochemical properties of mango wine with maceration and enzyme treatment 110 9.2.2 Volatiles of mango wine with pulp contact and enzyme treatment .112 vii 9.2.2.1 Terpenols and monoterpene hydrocarbons .112 9.2.2.2 Alcohols 116 9.2.2.3 Esters and fatty acids 117 9.2.2.4 Minor volatile compounds 117 9.2.2.5 Sensory characteristics of mango wine .118 9.2.2.6 Principal component analysis of volatiles in mango wine 120 9.3 Conclusion .121 CHAPTER 10 General conclusions and future work .122 REFERENCES .125 viii Hansen, H. 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Journal of Food Composition and Analysis, 10(1), 55-65. 146 [...]... chromatogram of (E)-β-damascenone in non-macerated control, macerated wine control, non-macerated with enzyme and macerated wine with enzyme 118 Figure 9.2 Aroma profile of mango wines: non-macerated control (▲); macerated wine control (■); non-macerated with enzyme (♦); and macerated wine with enzyme (× 119 ) Figure 9.3 Biplot of principal component analysis of mango wines: nonmacerated control... acid and sugar concentrations of mango wine 47 Table 4.2 Summary of volatiles in mango juice and wine with 21-day fermentation 48 Table 4.3 Summary of changes of major terpenoids and their derivatives from mango juice (day 0) to mango wine after 21-day fermentation 50 Table 4.4 Concentrations of odorants (mg/L) and their corresponding odor activity values (OAVs) in mango wine fermented... cerevisiae and W mrakii 72 Table 6.2 Categorization of volatiles of fresh mango juices (Day 0) and mango wine (Day 21) from three varieties 76 Table 6.3 Concentration, odor thresholds, odor activity values (OAVs) and odor description of potent odorants in mango wines 78 Table 8.1 Physicochemical properties, alcohol, organic acid, and sugar concentrations of mango wines with and without... 8.2 Volatiles (mg/L) and their odor activity values (OAVs) of nonmacerated and macerated mango wines 103 Table 9.1 Physicochemical properties, alcohol, organic acid, and sugar concentrations of non-macerated and macerated mango wine, with and without enzyme treatment 112 Table 9.2 Major volatiles (mg/L) and their odor activity values (OAVs) for non-macerated and macerated wine, with and. .. the senses of taste and smell of the consumer Wine flavor is composed of non -volatile chemicals which lead to taste sensations and volatile compounds which are responsible for the odor The non-volatiles include sugars, organic acids, polyphenols, etc., which cause sweetness, 5 sourness and astringency, respectively The concentration of non-volatiles needs to be at level of ~1% to influence wine taste... varieties of mangoes from India were screened for production of dessert and madeira-style wine and tested for influence of madeirization on organoleptic quality (Onkarayya & Singh, 1984) Obisanya et al (1987) studied the fermentation of mango juice into wine using locally isolated yeasts Saccharomyces cerevisiae and Schizosaccharomyces species of palm wine and they concluded that Schizosaccharomyces yeasts. .. 20)α-terpineol 21)β-citronellol 22)β-damascenone 23) γoctalactone 24) β-Ionone 25) δ-octalactone 26) γ-decalactone 27) δnonalacton 75 xiv Figure 6.4 Evolution trend of terpene hydrocarbons and β-citronellol throughout the fermentation of mango juices of three varieties: „R2E2‟ (♦),‟Harum Manis‟ („HM‟) (▲) and „Nam Doc Mai‟ („NDM‟) (■) 77 Figure 6.5 Evolution trend of (Z)-3-hexenol and isoamyl... the production of sweet, table mango wine and Saccharomyces yeasts were suitable for the production of dry mango wine with a higher ethanol level (Obisanya, Aina, & Oguntimein, 1987) Some recent studies about mango wine started in India in 2005 A method of mango juice extraction with pectinase was developed Ethanol as well as some volatile contents of mango wine was characterized It was concluded that... yeast strains on day 21 54 Table 5.1 Changes of sugars, organic acids, ethanol and glycerol in mango wines before and after fermentation 59 Table 5.2 Concentrations, odor thresholds, odor activity values (OAVs) and odor description of typical odorants in mango wines 62 Table 6.1 Changes of sugars, organic acids, ethanol and glycerol in mango wines before and after fermentation of three varieties... temperature, pH and inoculum levels of monocultures of S bayanus to optimize the production of ethanol, glycerol, and volatile acidity They reported that the predicted value for optimisation process conditions were in good agreement with experimental data (Kumar, Prakasam, & Reddy, 2009) Furthermore, influence of condition factors (temperature, pH, SO2, and aeration) on mango wine fermentation was further . IMPACT OF SELECTED YEASTS AND GLYCOSIDASE ON CHEMICAL COMPOSITION OF MANGO WINE LI XIAO NATIONAL UNIVERSITY OF SINGAPORE 2013 IMPACT OF SELECTED YEASTS AND GLYCOSIDASE. 3.2.4.4 Carbonyl compounds 43 3.3 Conclusion 44 CHAPTER 4 Impact of two Williopsis yeast strains on the volatile composition of mango wine 45 4.1 Introduction 45 4.2 Results and discussion 45. glycerol contents of mango wine 98 8.2.4 Volatile profile of mango juice and wine 99 8.2.4.1 Major volatiles in juice 99 8.2.4.2 Evolution of terpenes hydrocarbons 99 8.2.4.3 Evolution of alcohols