THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY Mohammad Mazbah Uddin Bahar A STUDY ON THE EXTRACTION OF POLYSACCHARIDES FROM GANODERMA LUCIDUM BACHELOR THESIS Study mode: Full-time Major: Food Technology Faculty: Advanced Education Program Office Batch: 2018 - 2022 20th February 2023 1 Degree Program Bachelor of Food Technology Student name Mohammad Mazbah Uddin Bahar Student ID Thesis Title A study on the extraction of polysaccharides from Ganoderma lucidum Supervisor (s) Dr. Luong Hung Tien Supervisor’s Signature Abstract: Ganoderma lucidum is a pharmaceutical fungus that offers plentiful bioactive compounds, which take part in enhancing human’s health. This study aimed to provide more scientific insight toward the extraction of the fungus, thus creating a new foundation for further development of specialized products based on G. There were 3 objectives, corresponding to 3 research contents.
The first was to determine the moisture and the ash contents of the input material. In this study, the moisture content of G.51% while the ash content was 2.06% of the dry weight. For extraction, this study employed the ultrasonically-assisted extraction (UAE) and examined the best operational conditions for extracting G. Optimization was conducted using Response Surface.
The results were an ultrasonically-processing time of 15 minutes, a temperature of 68oC, time of 158 minutes, and the solid-liquid ratio of 1:28 g/ml with ethanol 80% as the primary solvent. This operation produced 1.55% of polysaccharides extracted from the fungus. Keywords: Ganoderma lucidum, fungal extraction, polysaccharides Number of pages: 60 Date of Submission: i Acknowledgment The accomplishment of this graduation thesis is one of the greatest achievements that I have had in years. However, it came with much effort and the precious enthusiasm from not only myself but also others.
Thank you, Doctor Luong Hung Tien, for being my supervisor and my lecturer. You have always been intelligent, visionary, and you express that in really cool styles, which I love and take much inspiration from. I wish you strength, success, and all the best in the future, sir. Many thanks to the lecturers who have been teaching me and my friends.
Without your lessons and your thorough guides, I am certain that I would not have been able to work on not just this, but also my future. The faculty of Advanced Education Program definitely helped me a lot with my progress. Thanks to their schedules and information, I was able to arrange the work, which led to not just this thesis but also the completion of other plans. I am grateful for that.
May we all be blessed! Muhammad Mazbah Uddin Bahar ii TABLE OF CONTENTS I. The research subject – Ganoderma lucidum. Overview of Ganoderma lucidum. Chemical composition and bioactivity.
Pharmacological effects on human health. Usage on the global scale. Oriental herbal therapies. Modern medicine and pharmacy.
Extraction and compound isolation techniques. Solid-Liquid extraction. Advanced extraction techniques. Experiment design and methods.
The quality of input materials. The extraction of Ganoderma lucidum. RESULTS AND DISSCUSION. Input indexes of Ganoderma lucidum.
The effects of environmental factors on extracting G. Ultrasonically processing time. Type of solvent. Solid – liquid ratio.
Optimal extraction conditions for G. Ultrasonically processing time. Types of solvent. Solid-Liquid ratio.
57 iv LIST OF TABLES Table 3. Chemicals involved in the process. Investigated range for ultrasound. Investigated types of solvent.
Investigated concentrations of solvent. Investigated temperature levels. Investigated range of extraction time. Investigated solid-liquid ratios.
Dilution formulas of the control solution. Response surface method’s design and investigated parameters. Experiment design for optimization experiments. Moisture content and ash content of G.
Total polysaccharides from different ultrasonically processing time. Total polysaccharides from different types of solvent. Total polysaccharides from different solvent concentrations. Total polysaccharides from different temperatures.
Total polysaccharides from different time. Total polysaccharides from different solid-liquid ratio. Input values of optimization matrix. 36 v LIST OF FIGURES Figure 3.
Standard curve of diluted glucose. Polysaccharides from ultrasonically-processing time. Polysaccharides from types of solvent. Polysaccharides from solvent concentration.
Polysaccharides from temperatures. Polysaccharides from extraction time. Polysaccharides from solid-liquid ratios. Rationale Ganoderma is a common name of various fungi that are relatively popular as a pharmaceutical ingredient in Asian countries, especially Eastern – Southeast Asia.
Particularly, they take part in herbal therapies (Hijikata et al., 2005; Shanshoury et al. The number of species in genus Ganoderma is also impressive with notable names such as G. Depending on the specific species, there are distinct profiles of chemical composition and bioactivity, thus the pharmacological effects vary accordingly (Du et al. That is why comprehension over these subjects or one of them in particular is much needed for using them in the most proper way.
Despite much appreciation and acknowledgement throughout the communities of medicine and pharmacy, Ganoderma species, particularly Ganoderma lucidum, have not been utilized up to their potential. In modern countries, there are several specialized products taking form as dietary supplements such as capsules (Wicks et al., 2007; Rossi et al. Many respected pharmaceutical companies in China have developed products with health effects of blood and heart regulation, anti-fatigue and depression, as well as anti-cancer and tumor. Nevertheless, there are still many markets remaining for the development of a specialized product from Ganoderma lucidum like Russia and Bangladesh, where this material is relatively uncommon.
Even in Southeast Asian countries like Vietnam, the use of Ganoderma lucidum has only been limited with the raw form infused in hot water. This technique ultimately leads to inefficient extraction, thus resulting in waste in terms of material and cost. From these facts, it is comprehensible that such a miraculous material as Ganoderma lucidum needs to be further promoted through studying and developing. That is why the study on “The extraction of Polysaccharides from Ganoderma lucidum” was implemented.
This study promised to examine the Ganoderma lucidum 1 in terms of the total polysaccharides as well as the proper extraction method in order to optimize both the operation and the cost. Research hypotheses Based on the rationale and the literature review, there were the following hypotheses set for the subject of Ganoderma lucidum: 1. Ganoderma lucidum can be further promoted and disseminated in other regions like Russia, Bangladesh, and Vietnam. There is a way to specialize G.
lucidum instead of consuming it daily as water infusion 3. Optimization is required to produce the most efficient extraction process while saving the cost. From those hypotheses, questions were made and information was required to answer the practicality, novelty, and topicality of this study. The objectives and the work frame of this study were also inferred during the process of gathering literature bases.
Research objectives This study had 3 primary objectives as follows: • Identify the quality of input Garnoderma lucidum • Identify the technique for extraction as well as conditions in the technique • Optimize the extraction process, particularly the operational conditions These objectives led to the research contents of the study. Limitations Research capability and practice: Capability exists in not only instruments, techniques, and protocols but also in humans, who are responsible for executing the research. Such a limit, although it could be minimized in many ways, still remains to a certain extent and thwart achievements at higher levels. 2 Literature inheritance: It is expected from any scientific project to employ the results of others to discuss and further fortify that project.
However, there are many reasons that make such an act a limitation. Prominently, the variability in terms of practice, equipment, and materials often results in values that possibly contradict the findings of others. Another reason is that the accessibility to scientific resources is limited, which leads to less information and discussion. The research subject – Ganoderma lucidum.
Overview of Ganoderma lucidum. The classification of Ganoderma fungi depends on the color of the species and lucidum is the species with the red color. It is possibly the most commonly found variation and is considered as the best type in terms of bioactivity. According to the classification, biological characteristics, and cultivations of Ganoderma documented by Du et al.
(2019), Ganoderma lucidum is a fungus with a complete compositional structure like a regular one, which means there are the mycelium, the stem, the ring, the gill, the cap, and the spore. Although the mycelium and the spore of G. lucidum are deemed usable with various beneficial bioactive compounds, the fruiting body consisting primarily of the cap and the stem is the most popular part. Unlike a regular fungal body, the fruiting body of G.
lucidum is relatively hardened, sharing similarities with a wooden structure rather than a soft and limp body. This part protrudes out front and accounts for most of the full proportion of an individual. That is why it is easier to harvest and also offers more benefits. Kingdom: Fungi Phylum: Basidiomycota Class: Aragicomycetes Order: Polyporales Family: Ganodermaceae Genus: Ganoderma Species: lucidum Figure 1: A Ganoderma lucidum’s fruiting body.
Credit: BGN 4 As recorded in the Medicinal Journal of Wachtel-Galor et al. (2011), Ganoderma lucidum grows wildly in mainly China, Japan, Korea, some Southeastern and Eastern Asian countries like Vietnam, Laos, Nepal, and a small part of Eastern Europe. This is also observed in other studies regarding the natural distribution of the fungus ( Jin et al., 2012; Du et al. That is likely the reason why G.
lucidum appears more popular in these regions instead of others. The study of Loyd et al. (2018) also found wild G. lucidum in North America.
However, it was possibly the product of migration. Chemical composition and bioactivity. Triterpenoids The group of compounds that makes Ganoderma lucidum original is the triterpenoids within the fruiting body. According to Lee et al.
(2010), the most prominent names are lanostane triterpenoids, which are virtually not present in other family than Ganoderma. This study had also identified 18 different lanostane with 4 new compounds of butyl ganoderate A, butyl ganoderate B, butyl lucidenate N, and butyl lucidenate A. Another study of Lin et al. (2022) on the triterpenoid profile of GL’s fruiting body also revealed 17 different lanostanes as well as 4 new triterpenoids with various health effects.
The difference between the two studies proved that the triterpenoids within Ganoderma lucidum can vary according to the origin and the cultivation environment. Polysaccharides Polysaccharides in Ganoderma lucidum are also renowned for their diversity and potentials. The profile of polysaccharides in Ganoderma lucidum is vast and even remains with undiscovered groups of compounds. Nevertheless, there are beta- glucans, which are the representative for GL’s polysaccharides (Benkeblia, 2016).
The components of this group are often mannose, xylose, glucose, galactose, and arabinose. They form a backbone of beta-1-3-D-glucopyranosyl and various branches 5 (mono-, di-, tri-). The study of L. Li et al.
(2018) even compared the polysaccharide of Ganoderma lucidum and Ganoderma sinense. This direction had achieved significance in terms of revealing the vast profile of polysaccharides within the fungus. From that, it was possible to systematically speculate various health effects ranging from macrophage effects, tumor-suppressive effects, anti-inflammation, etc. Microelements There are also other nutrients and constituents that contribute to the bioactivity of Ganoderma lucidum.
Minerals, for example, are extremely abundant in a fruiting body. According to Wachtel-Galor et al. (2011), the fungus is rich in potassium, magnesium, calcium, sulfur, and phosphorus. There are also minor minerals like iron, copper, sodium, and zinc.
The vitamins in G. lucidum are also notable with original types like riboflavin, niacin, and thiamin (El Sheikha, 2022). Pharmacological effects on human health. With a diverse and plentiful profile of beneficial constituents and bioactive compounds, Ganoderma lucidum offers numerous health benefits for the consumer.
In terms of regular uses, the fungus was reported with abilities to enhance the immune system. The study of Zhao et al. (2018) on the interaction between G.