MINISTRY OF EDUCATION AND TRAINING HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY FOR HIGH QUALITY TRAINING GRADUATION THESIS FOOD TECHNOLOGY ENZYMATIC HYDROLYSIS OF GENIPOSIDE FROM GARDENIA JAMINOIDE TO PRODUCE GENIPIN AS A PIGMENT PRECURSOR AND CROSSLINKING AGENT SUPERVISOR: VO THI NHA NGUYEN VINH TIEN STUDENT: DANG HOANG DUC HO DAC LOC SKL 0 0 9 1 5 2 Ho Chi Minh City, August, 2022 HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY FOR HIGH QUALITY TRAINING GRADUATION PROJECT Thesis code 2022-18116009 ENZYMATIC HYDROLYSIS OF GENIPOSIDE FROM GARDENIA JAMINOIDE TO PRODUCE GENIPIN AS A PIGMENT PRECURSOR AND CROSSLINKING AGENT DANG HOANG DUC Student ID: 18116009 HO DAC LOC 18116022 Major: FOOD TECHNOLOGY Supervisor: VO THI NGA, PhD. NGUYEN VINH TIEN, ASSOC. Ho Chi Minh City, August 2022 HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY FOR HIGH QUALITY TRAINING GRADUATION PROJECT Thesis code 2022-18116009 ENZYMATIC HYDROLYSIS OF GENIPOSIDE FROM GARDENIA JAMINOIDES TO PRODUCE GENIPIN AS A PIGMENT PRECURSOR AND CROSSLINKING AGENT DANG HOANG DUC Student ID: 18116009 HO DAC LOC 18116022 Major: FOOD TECHNOLOGY Supervisor: VO THI NGA, PhD. NGUYEN VINH TIEN, ASSOC.
Ho Chi Minh City, August 2022 DECLARATION Except where there is clear acknowledgment and reference to the work of others, we thus declare that all content and materials included in and presented in this thesis are our original creations. Additionally, we guarantee that the materials acknowledged in the thesis have been cited appropriately and properly in line with requirements. , August 2022 Signature I ACKNOWLEDGEMENT "No research without action, no action without research" is a well-known quote attributed to Kurt Lewin (2012). Indeed, in order to finish and attain the current outcome, we have addressed and conquered all the difficulties and obstacles inherent in and throughout the project.
In addition, our beloved professors, classmates, and families have made a substantial contribution that stimulates and supports us. Therefore, we would like to sincerely thank all the lecturers in charge of the Department of Food Technology, Faculty of Chemical and Food Technology, and Ho Chi Minh City University of Technology and Education for providing us with valuable knowledge and the best equipment and facilities to complete our thesis. We would also want to express our gratitude to our cherished supervisors, PhD. Vo Thi Nga and Assoc.
Nguyen Vinh Tien, who have passionately guided and shared their teaching expertise and experience in order for us to complete this thesis. Sincerely, we would like to thank Ms. Ho Thi Thu Trang of the Department of Food Technology for allowing and assisting us in using the available measuring instruments and equipment at the Faculty of Chemical and Food Technology laboratory. Nonetheless, we would want to express our gratitude to our classmates for supporting and organizing the thesis's complex experiment.
II III IV V VI VII VIII IX X XI XII TABLE OF CONTENTS CHAPTER 1: OVERVIEW. Volatile components in G. Iridoids and iridoid glycoside. Crocins and their derivatives.
Effects of blood circulation. Reasearch about gardenia blue pigment production. Cross-linking of genipin in chitosan film. Reasearch about forming crosslinking with genipin .13 CHAPTER 2: MATERIAL AND METHOD.
Research process diagram. Pigments from genipin processing. Extracting geniposide from seed of G. Jasminoides in ethanol.
Treating geniposide with cellulase to obtain a hydrolysate. Extracting genipin from the hydrolysate by ethyl acetate. Reacting the product comprising genipin with amine. Chitosan-genipin film processing.
Procedure to investigate the optimal pH of the enzymatic hydrolysis of geniposide. Procedure to investigate the optimal duration of the enzymatic hydrolysis of geniposide. Procedure to investigate the optimal enzyme concentration of the enzymatic hydrolysis of geniposide. Procedure to investigate the optimal pH of the reaction between genipin and pigment precursors.
Procedure to investigate the optimal duration of the reaction between genipin and pigment precursors. Procedure to investigate the different amine of the reaction between genipin and amine. Procedure to investigate the optimal pH of the reaction between genipin and protein extracted from Phaseolus lunatus. Procedure to investigate the differences of chitosan-genipin films in different genipin concentration.
FTIR – Fourier Transform Infrared Spectroscopy. Tensile strength and elongation .25 CHAPTER 3 : RESULTS AND DISCUSSION. Factors affecting the hydrolysis reaction of geniposide. Time of enzymatic reaction.
Factors affecting the reaction to produce blue pigments from genipin. Time of pigment forming reaction. Types of amine-containing compounds. Effect of pH on protein of Lima bean when reacting with genipin.
Uv-vis of pigment solution when genipin act with protein from Phaseolus lunatus in different pH. FTIR of residue of proteinwhen acting with protein from Phaseolus lunatus in different pH. Properties of chitosan-genipin films. Uv-vis of films when change genipin content.
FTIR of films when change genipin content. Moisture content of genipin – chitosan films (%). Mechanical properties of genipin – chitosan films (Thickness, TS and EL) 50 3. Swelling content of genipin – chitosan films.
53 XV LIST OF FIGURES Fig. Structure of crocin. The structural formula of geniposide .Hydrolysis of geniposide to genipin [29]. Chemical structure of genipin.
Garnedia blue reaction between genipin and primary amines. Crosslinking reaction between chitosan and genipin. Pigments from genipin producing diagram. Chitosan-genipin film producing diagram.
Gardenia blue solution in different pH of enzymatic reaction. (a) UV spectra of genipin solution after enzymatic reaction in different pH of enzymatic reaction. (b) Absorbance of 310 nm of genipin solution after enzymatic reaction in different pH of enzymatic reaction. (a) UV-vis spectra of genipin solution after ethyl acetate extracted in different pH of enzymatic reaction.
(b) Absorbance of 310 nm of genipin solution after ethyl acetate extracted in different pH of enzymatic reaction. (a) UV-vis spectra of gardenia blue pigment in different pH of enzymatic reaction. (b) Absorbance of 590 nm of gardenia blue pigment in different pH of enzymatic reaction. Gardenia blue solution in different time of enzymatic reaction.
(a) UV-vis spectra of genipin solution after enzymatic reaction in different time of enzymatic reaction. (b) Absorbance of 310 mn of genipin solution after enzymatic reaction in different time of enzymatic reaction. (a) UV-Vis spectra of genipin solution after ethyl acetate extraction in different time of enzymatic reaction. (b) Absorbance of 310 nm of genipin solution after ethyl acetate extraction in different time of enzymatic reaction.
(a) UV-Vis spectra of gardenia blue solution in different time of enzymatic reaction. (b) Absorbance of 310 nm of gardenia blue solution in different time of enzymatic reaction. The content change of geniposide and genipin determined by HPLC [63]. Gardenia blue solution in different enzyme concentration.
(a) UV-Vis spectra of genipin solution after enzymatic reaction in different enzyme concentration. (b) Absorbance of 310 nm of genipin solution after enzymatic reaction in different enzyme concentration. (a) UV-Vis spectra of genipin solution after ethyl acetate extracted in different enzyme concentration. (b) Absorbance of 310 nm of genipin solution after ethyl acetate extracted in different enzyme concentration.
(a) UV-vis spectra of gardenia blue solution in different enzyme concentration. (b) Absorbance of 310 nm of gardenia blue solution in different enzyme concentration. Gardenia blue solution in different pH of the formation of garnedia blue reaction. (a) UV-vis spectra of gardenia blue solution in different pH of the formation of garnedia blue reaction.
(b) Absorbance of 590 nm of gardenia blue solution in different pH of the formation of garnedia blue reaction. (a) UV-vis spectra of gardenia blue solution in different time of the formation of garnedia blue reaction. (b) Absorbance of 590 nm of gardenia blue solution in different time of the formation of garnedia blue reaction. Color difference of solution after reacting with different amines.
UV-vis spectra of gardenia blue solution in types of amine-containing compounds. Color difference of reaction between genipin and Phaseolus lunatus extraction in different pH. (a) UV-vis spectra of solution when genipin act with protein from Phaseolus lunatus in different pH. (b) Absorbance of 590 nm of when genipin act with protein from Phaseolus lunatus in different pH.
FTIR of protein residue after being treated with genipin. UV-vis spectra of genipin-chitosan films. FTIR of genipin-chitosan films .48 XVIII LIST OF TABLES Table 1. Composition of Crocin colorant in gardenia fruit.
Preparation of different amine solutions. Buffer solution preparation. Color measurement of gardenia blue solution in different pH of enzymatic reaction. Color measurement of gardenia blue solution in different enzyme concentration.
Color measurement of gardenia blue solution in different pH of the formation of garnedia blue. Color measurement of gardenia blue solution in different time of the formation of garnedia blue reaction. Color measurement of gardenia blue solution in different amines. Moisture content of genipin – chitosan films.
Mechanical properties of genipin-chitosan films. Swelling content of genipin – chitosan films .51 XIX ABSTRACT Gardenia blue pigment is typically made from the raw material geniposide found in Gardenia Jasminoides Ellis of Rubiaceae by processing geniposide with α-glucosidase to create genipin, which then combines with an amino acid to form the color. However, the resulting gardenia blue pigment is dark, has a low color value, and is of poor quality. Therefore, it is unsuitable for some uses, including drinks.
A second method for producing the gardenia blue pigment with a high color value involves ultra- filtering the gardenia blue pigment obtained from the reaction of genipin with an amino acid to remove the residual geniposide and then extracting the filtrate to obtain the high-color-value gardenia blue pigment. Another method involves putting the raw material geniposide through a non-polar resin with a wide mesh to remove α-crocin prior to treating it with β-glucosidase. However, due to the high cost and complexity of these procedures, they cannot be carried out on a wide scale in the industry. This study, focuses on the pH sensitive, enzyme concentration and time of the enzymatic reaction between geniposide and cellulase, pH sensitive, type of pigment precursors, time of the pigment forming reaction.
And pH sensitive of reaction between genipin and protein from Lima bean. The result for producing pigment from Gardenia jasminoides are pH 4.2 g cellulase per 1 g geniposide, 6 hours are the optimal conditions for the enzymatic reaction between geniposide and cellulase. The evaluated method for this research are UV-vis and color measurement. Moreover, pH 8, MSG, 10 hours are the optimal conditions for the pigment forming reaction.
The evaluated method for this research are UV-vis and color measurement. Additionally, pH 10 is the optimal condition for the reaction between genipin and protein from Lima bean. Regarding the genipin-chitosan film, when changing the genipin/NH2 concentration, the crosslinking between chitosan and genipin also changed. In addition, the concentration of 0.0075 genipin/NH2 had the highest tensile strength.
Moreover, adding concentration also causes moisture and swelling to decrease gradually. Keywords: Gardenia jasminoides, genipin, geniposide, gardenia blue, crossliking, genipin- chitosan films. XX CHAPTER 1: OVERVIEW 1. jaminoides, an evergreen tree of the Rubiaceae family, is planted in many parts of China under the Chinese name Zhi Zi.
It thrives in a variety of temperate climates and has fragrant white blooms [1]. When the plant's oval-shaped fruits ripen in late fall, they become a reddish- golden hue [2]. jaminoides possess a variety of biological functions, including anti-diabetic, anti- inflammatory, antidepressant, and antioxidant qualities, as well as the ability to improve sleep quality [3].jasminoides herb has the ability to access the meridians of the heart, lungs, and triple burner. It has the ability to extinguish an evil fire, ease internal heat, and cool blood in the body.
It is mainly used to treat dysphoria, agrypnia, jaundice, gonorrhea, thirst, conjunctival congestion, angina, hematemesis, non-traumatic bleeding, hematodiarrhoea, hemuresis, pathopyretic ulcer, sprain, and swelling pain [4]. According to recent studies, the oil extract of Gardenia jasminoides has antidepressant properties [5]. It has long been used as a natural yellow dye.