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thesis 4.8 corrected

thesis 4.8 corrected
thesis 4.8 corrected

Poznań University of Life Sciences

Faculty of Food Science and Nutrition

Major: Food Science and Nutrition

Yuan Xindi

(album nr)

Research on the Antioxidant Activity of Series Kombucha

The MSc thesis made

at the Faculty of Food Science and Nutrition

Research work done under the supervision of

dr hab. Anna Gramza-Michalowska, prof. nadzw.

at the Department of Food Service and Catering

Thesis accepted…………………………………

Poznań 2016

I am very grateful to my supervisor, Prof. Anna Gramza-Michalowska, for teaching and helping me design the subject of my research and thank to my supervisor I improved a lot during finishing this project. She gives me not only the knowledge how to solve the problem during the research and inspiration, also the spirit of perseverance.

Abstract:

Effect of culture time on the antioxidant capacity of kombucha beverage was studied. Research included the determination of DPPH radical scavenging ability and the total polyphenol content of different types of kombucha beverage. Study allowed to compare the variation of antioxidant capacity with cultivation time of four types of kombucha beverage and pure tea leaves beverage. Analysed beverages were also evaluated according to its sensory value. Experiments results showed that yellow kombucha beverage own the highest antioxidant capacity evaluated with the 1,1-diphenyl-2-pierylhydrazyl (DPPH) radical scavenging ability, as well as the total phenolics content in comparison with other tea samples. (2) Results of sensory evaluations of kombucha teas indicated acceptable performance in four types of kombucha, especially yellow tea leaves kombucha.

Keywords: kombucha beverage, tea leaves, Camellia sinensis, antioxidant capacity, sensory evaluation.

Table of contents

1. INTRODUCTION ................................................................... 错误!未定义书签。

1.1. The type and strain of bacteria Kombucha ....................... 错误!未定义书签。

1.1.1.Yeast. (3)

1.1.2.Xylinum (Acetobacter Xylinumm) (4)

1.1.3. Kombucha layer (5)

1.1.4. Lactic acid bacteria (7)

1.2. Antibacterial effect of Kombucha (8)

1.3. Study of fermentation conditions (9)

1.4. Kombucha component analysis (10)

1.5. Anti-Oxidation of kombucha fermented tea (11)

1.6. Mechanism of Action (12)

1.7. Production Method for different kinds of tea (14)

1.7.1. Green tea (14)

1.7.2. Yellow tea (15)

1.7.3. White tea (15)

1.7.4. Black tea (16)

1.8. Aim of research (17)

2. MATERIALS and METHODS (17)

2.1. Devices and reagents (17)

2.1.1. Main material (17)

2.1.2. Main reagents (18)

2.1.3. Main devices (18)

2.2. Experiment method (18)

2.2.1. Kombucha fermentation production (18)

2.2.2. Determine DPPH free radical antioxidant activity (19)

2.2.3. Folin-Ciocalteu Colorimetric method (21)

2.2.4. Determine dry mass (22)

2.2.5. Kombucha sensory evaluation method..................................................

2 2

3. RESULT and ANALYSIS (24)

3.1. Comparison of Different Types of Kombucha Ability for DPPH Free Radical Scavenging (25)

3.1.1. Determine the Dilution (25)

3.1.2. Relations between DPPH Solution Absorbance and Concentration. (25)

3.1.3. Effects of Different Kinds of Teas on the Kombucha for DPPH Free

Radical Scavenging Capacity. (26)

3.1.

4. Effects of Different Cultural Time on the Kombucha for DPPH free

radical scavenging (33)

3.1.5. Effects of Different Type of Tea Leaves on Kombucha for DPPH free

radical scavenging during storage (36)

3.2. Determination of Total Polyphenol in Kombucha (37)

3.2.1. Relations between Absorbance and Total Polyphenol Content The

Standard Curve. (38)

3.2.2. Effects of Different Kinds of Tea on the Kombucha for The Total

polyphenol Content by Folin—Ciocalteu assay (39)

3.3. Comparison of different types of Kombucha drink to effect for dry mass (43)

3.4. Sensory Evaluation of Different Kinds of Tea Samples (45)

4. CONCLUSION (51)

REFERENCES (53)

1. INTRODUCTION

Kombucha has a long history of folk tradition of acidic beverages. It is based on sugar tea as raw material, acetic acid bacteria, yeast and lactic acid bacteria and other microorganisms common fermented. Over the past decade, Kombucha very popular in some European and American countries and regions, for a variety of health function in kombucha is ascendant, especially its detoxification cancer and enhance immunity and other functions concern, increasingly the more data indicate that Kombucha is a very valuable use of natural microbial fermentation products. Kombucha Although China has a long history, but its scientific research has started late. Article that Kombucha microbial species mainly acetic acid bacteria and yeast, while noting that the acetic acid bacteria Kombucha more than one, but a film based vinegar bacteria. The late 1970s to the early 1980s, Kombucha drink was very popular in China, some introductory Kombucha health effects and production methods of articles in large numbers ridicule (Guo,2007). Early 20th century to the mid, some doctors and researchers have reported kombucha, but the study of bacteria and the mechanism of action is still very few. In recent years, China's anti-bacterial effect of Kombucha on a number of studies. From the mid-20th century until now, about Kombucha microbial composition and interaction of research fermentation conditions, bacteria composition, mechanism and other aspects of the succession has been reported. The present study showed that the presence of antibacterial proteins in Kombucha tea. It can be used as food preservative extracted to replace chemical preservatives. As for the extraction of the antifungal protein and its application in food preservation will be the future research directions. Kombucha tea contains some antibacterial ingredients (such as organic acids, etc.), Kombucha tea Escherichia coli(G.), Bacillus subtilis(G +), Staphylococcus aureus(G +) has a certain extent. Until now, research mainly direction on segregation Kombucha strain and identification bacteria composed of

metabolites(organic acids and alcohols).

It is sugar tea as raw material, acetic acid bacteria, yeast and lactic acid bacteria and other microbes fermented together. Kombucha tea extract containing a portion of tea, live microorganisms and their metabolites, these substances include glucose acid, acetic acid, glucose, fructose, proteins, amino acids, vitamins, trace elements, polyphenols, caffeine, alcohol and carbon dioxide (Blanc,1996).That has been drink Kombucha It is considered to be a variety of health effects on human body. Its health effects include: clean up the stomach, prevention and treatment of constipation and hemorrhoids; help digestion; for hypertension, hyperlipidemia, atherosclerosis and other cardiovascular diseases and diabetes prevention and treatment; to avoid the occurrence of a variety of stones; helps the body in a timely manner eliminate toxins, anti-cancer, anti-cancer and enhance immunity and so on (Guo.2007). Overall, many kombucha human health effects are the result of a variety of beneficial ingredients working together.

1.1 The type and strain of bacteria Kombucha

Kombucha strains for fermentation mainly acetic acid bacteria and yeast, and some have a small amount of Kombucha Lactobacterium bulagricum(Xie, She ,2000). So far, the separation of people from a variety of Kombucha to acetic acid bacteria, the most important is Xylinum (Acetobacter Xylinumm) and yeasts. Kombucha fermenting bacteria from one or more of the above acetic acid bacteria and yeast one or more of the above composition, and some have lactic acid bacteria. Different species composition, type and quantity of bacteria in its metabolites will be different. Because of its species diversity, Kombucha is complex metabolic processes. Yeast and acetic acid bacteria is mutually beneficial symbiotic relationship in Kombucha. At the start of the fermentation stage, the acetic acid bacteria can not directly use sucrose or sucrose utilization rate is very slow, the yeast will degrade sucrose into glucose and

fructose, and further fermented to produce ethanol, acetic acid bacteria in the culture medium with glucose, fructose and after the growth and reproduction of large quantities of ethanol, glucose and fructose, glucose oxidation metabolites acid, acetic acid, etc., and oxidation of ethanol produced by yeast to produce acetic acid. It plays an important role in the early stages of the fermentation of yeast reproduction. Data show that yeast produce ethanol can stimulate the growth of acetic acid bacteria to produce more cellulose membrane and acetic acid and acetic acid bacteria produce will stimulate yeast to produce ethanol, and acetic acid, the presence of ethanol to acetic acid bacteria protection and yeast to make them from other microorganisms infection (BLANC,1996). This is also an important reason Kombucha has a strong antibacterial activity.

1.1.1Yeast

Yeast cells are rich in protein and vitamins, so it can be added to foods to make senior nutrition or used as advanced feed for animal feeding. Yeasts are single-celled microorganism, its taxonomic status and molds, as are all fungi, except that the mold is a unicellular or multicellular filamentous fungus. Its size several times larger than the Acetobacter Xylinumm, so it is easy to observe the separate area with Acetobacter Xylinumm under a microscope. Different yeasts and bacteria, it prefers to grow in acidic conditions, the optimum pH its growth is 4.5 to 5.5. It can multiply and produce many metabolite which is beneficial to humans, e.g., vitaminBl, vitaminB2, citric acid, succinic acid and other organic acids, amino acids and proteins, fats, nucleic acid substances, various hormones under aerobic conditions (Gu,1981). There are many types of yeast in the case of oxygen, glucose and other sugars can be fermented into alcohol and CO2, at the same time grow slowly. Most yeast can utilize glucose, fructose, sucrose carbon source, it is possible to make use of ammonium nitrogen, can not use nitrate. Although yeast can synthesize a variety of vitaminB, but

it is early in the growing medium must have vitaminB to grow quickly. Yeast optimum growth temperature is 25 oC ~ 30oC, the growth limit temperature is 4oC ~ 42oC . Yeast glucose metabolism can produce a variety of substances, lactic acid and acetic acid bacteria for the Xylinum provided nutrients of growth. Yeast is widely distributed in nature, especially like to grow in acidic environments, and more sugar, such as fruit, vegetables, honey, and the most common surface in the orchard soil. Yeast is a single cell fungi, bacteria oval, round or cylindrical, without flagella, can not move. Cell structure having the general characteristics of fungal cell, cell walls do not contain cellulose, dominated by dextran, polysaccharides such as mannan, also contains protein, lipids and chitin and other components. Yeast colonies circular, large, thick, mostly white, few red, showing grease-like shape or wrinkled skin. uniform turbidity in a liquid medium, some form precipitates ; some floating on the surface of biofilm yeasts can ferment various sugars assimilation or, generally glucose, sucrose, maltose, a few species can use five carbon sugars and starches in aerobic yeast. aerobic respiration and vigorous growth in anaerobic fermentation and slower growth rate (Gu ,1981).

1.1.2 Xylinum (Acetobacter Xylinumm)

Xylinum (Acetobacter Xylinumm) is a rod-shaped bacteria, single or short chain exists, it is aerobic bacteria, to be under aerobic conditions in order to rapid and massive growth. But it and other acetic acid bacteria is different under hypoxic conditions, it can use glucose anaerobic fermentation. The conversion of glucose into lactic acid and acetic acid, so it will not because of lack of oxygen and the large number of deaths quickly(Xie, 2000). Because of this feature, in addition to its Kombucha liquid culture medium in the case of surface oxygen quantity full blooms shaped pro biofilm, the bacteria can also be formed in the floe, to maintain a considerable number of cells. It is not resistant to drying in the culture medium or grow well under water and more

conditions. The optimum growth temperature is 28oC, limit the growth temperature 42oC, the mortality rate increased at a high temperature, Acetobacter Xylinumm at low temperatures is not easy to inactivation. Most bacteria like to grow under slightly alkaline conditions, but like acetic acid bacteria was grown under acidic conditions. Its optimum growth pH value is 5.4 to 6.0, 2.5 and 8.0 pH for growth limits. Acetobacter Xylinumm can utilize glucose, fructose, succinate and lactate as a carbon source; amino acids can be used as a nitrogen source; thiamine, niacin, amino acid and other vitamins wood acetic acid bacteria can promote. In aerobic conditions undergrowth acetic acid bacteria can quickly make the oxidation of glucose to gluconic acid, 2-keto-gluconate, oxidation of ethanol to acetic acid, so that the pH of the culture medium decreased rapidly and acidic; under hypoxic conditions, Acetobacter Xylinumm can convert glucose to lactic acid and acetic acid (Xie,2000). So, Kombucha liquid in fermentation process, Acetobacter Xylinumm play an important role in determine of bacteria quality.

1.1.3 Kombucha layer

BC (Bacterial cellulose) have become very popular biomaterials research in China and other countries. Synthetic hormone is generally considered plant-specific functions. However, a few organisms are able to synthetic hormone. A white fibrous substance Kombucha culture liquid surface is formed by a large number of many wood cellulose acetate synthesized in bacteria and other bacteria intertwined formation (Yamanaka,1989). By chemical and physical methods of analysis to identify such material has no structure and chemical properties of cellulose, so named because of its genus bacterial synthesis of bacterial cellulose. Xylinumm is acetic acid bacteria strains strongest synthetic fiber cable, which is quite amazing ability synthetic hormone. Acetic acid bacteria cell wall containing cellulose, and this is different from other bacteria, other bacterial cell wall is N- hexanoyl glucose as a

main component (Xie, 2000). Some recent studies have shown that an acetic acid bacteria cells per hour can put a million glucose units connected into cellulose, which can produce 3 ~ 4cm cellulose per day. Biosynthetic pathway of cellulose acetate is a bacterium a complex process. Bacterial fiber cable from glucose molecules β-1,4 glycosidic bonds into polymeric molecular chain molecule and intermolecular hydrogen bonds form a network structure, biosynthesis can be divided into aggregation, secretion, assembly and crystallization, four processes are highly coupled, and specific sites on the cell membrane are closely related (Jia,2001). Here is the four processes;

①Polymerization: Under the influence of glucokinase, the glucose is converted to glucose-6-phosphate;

②Secretion:Under the influence of isomerase, the glucose 6-phosphate is converted to glucose-1-phosphate;

③Assembly: Under the action of Uridine Diphosphate Glucose p yrophosphatase, the glucose-1-phosphate is converted to Uridine Diphosphate Glucose;

④Crystallization: Under the action of Cellulose Synthase, the Uridine Diphosphate Glucose is converted to β-1, 4 glycosidic bonds chain reassembled to form a cellulose. So Uridine Diphosphate Glucose to generate bacterial cellulose direct precursor. Bacterial cellulose acetate and synthetic bacterial cell growth between certain connection, therefore, can be improved by enhancing the growth of cellulose ability to grow bacteria. Cellulase is endoglucanase, a multi-enzyme complex extracellular glucosidase enzyme composition. But play an important role in cellulose synthesis enzyme activity is endoglucanase. Since A. Xylinum growth process itself will use glucose acid production when fermentation process proceeds, pH value of the medium

could be reduced, which is not conducive cell generation fibrinolytic. When adding acetic acid as a fermentation medium energy, continue to be consumed due to the acetic acid leaving the pH of the medium increased acid production and glucose consumption and therefore double the role of acetic acid to maintain a stable pH value of the culture medium, thereby facilitating cellulose production. Kombucha fermenting yeast in the early dominant flora, the first use of the fermentation of glucose to ethanol. The presence of bacteria increase the amount of ethanol in Acetobacter Xylinumm bacterial cellulose biosynthesis, biofilm forming bacteria shorten the time (Jia,2001).

1.1.4 Lactic acid bacteria

Lactic acid, similar in size, shape and acetic acid bacteria, it is not easy to distinguish between the two zones under a microscope. It grows not need oxygen, but a large amount of oxygen inhibition of the growth of lactic acid bacteria. Therefore, the lactic acid bacteria suitable for liquid layer growth, but not suitable for the growth of the liquid surface. Lactic acid bacteria can utilize glucose and lactose, convert them into lactic acid, but not using sucrose. Its growth needs of a variety of amino acids and vitamins. Lactic acid bacteria can produce interferon, interferon prevent the virus mRNA transcription and expression can inhibit the growth of cancer cells in vivo efficacy(Gu,1981).

Its mechanism of action in the following points: providing nutrients, can promote the growth of the body: to improve micro-ecological environment, clean intestines toxic substances; regulate digestion and the immune system. If the lactic acid bacteria play a normal metabolic activity in the body, it can directly provide essential amino acids (such as lysine and methionine, etc.) can be utilized as a host and various vitamins (vitamin B and K, H, etc.), but also improve mineral elements biological activity, and thus to provide essential nutrients to the host, enhanced animal nutrition metabolism,

direct and promote its growth (Gu,1981).

Kombucha fermentation strain is composed of one or more of the above acetic acid bacteria and yeast one or more of the above-mentioned composition, and some have lactic acid bacteria. Different species composition, type and quantity of bacteria in their metabolites will be different. Yeast and acetic acid bacteria is mutually beneficial symbiotic relationship in Kombucha. At the start of the fermentation stage, the acetic acid bacteria can not directly use sucrose or using rate is very slow, the yeast will degrade sucrose into glucose and fructose and further fermented to produce ethanol, acetic acid bacteria in the culture medium with the glucose, fructose and ethanol after the acetic acid bacteria start of a lot of growth and reproduction, glucose and fructose, glucose oxidation metabolites acid, acetic acid, etc., and oxidation of ethanol produced by yeast to produce acetic acid. Data show that yeast produce ethanol can stimulate the growth of acetic acid bacteria, produce more cellulose acetate membrane, but bacteria produce acetic acid will stimulate yeast to produce ethanol, and acetic acid, the presence of ethanol to acetic acid bacteria protection and yeast to make them from other microorganisms infection (Liu, Hsu,1996).

1.2 Antibacterial effect of Kombucha

Kombucha fermentation broth antibacterial substances in addition to the polyphenols in tea ingredients, also contain kombucha fermentation product. Many scholars believe that the kombucha antibacterial ingredients mainly from acid substances produced during fermentation, reportedly 1g/L concentration of acetic acid can inhibit the pathogenic bacteria and spore-forming bacteria (Wang,2015).

Kombucha in the presence of inhibitory substances also antibacterial protein produced by metabolic processes, and preliminarily determine the molecular mass greater than 3 500 u (Sreeramulu, Zhu,2000; Jiang, 2007); The study found that Lactobacillus may

secrete bacteriocins -plantaricin DSM20205, the bacteriocin is a strong thermal stability of small peptides, molecular weight 2 366.056 u, under acidic conditions of gram-positive bacteria and gram negative bacteria have significant inhibition (Zhang, 2008). To further determine the Kombucha in the antibacterial substance, the kombucha fermentation concentrate was adjusted to pH of 5.0, in order to eliminate the effects of acidic materials, at the same time 85oC heat treatment, measured antibacterial activity. This indicates that "Kombucha" there exist heat resistance antibacterial substance in fermentation broth, which may be Lactobacillus in the fermentation process synthetic bacteriocins and other substances, but the specific substance which is yet to be further studied.

1.3.Study of fermentation conditions

Effect of pH and total acidity of kombucha were studied on different temperatures (5 ~ 45oC) and a carbon source (sucrose, glucose, fructose and corn syrup), the results show acid fermentation optimum temperature is 30oC, similar types of organic acids with sucrose and fructose, dextrose and corn syrup produced. Fructose is much faster than the glucose metabolism, and metabolic rate than sucrose, fructose and glucose faster (Mi-Ae-Choi, 1994).

In the fermentation temperature of 20~22oC, and with different concentrations of different carbon sources (sucrose, fructose, lactose and glucose) of kombucha metabolites (ethanol and lactic acid) and the effect of pH do the research, the results show, sucrose and fructose to produce more ethanol, a small amount of ethanol is glucose. Sucrose can improve the output of lactic acid and lactic acid produced by other sugars rarely (Reiss,1994). The two species sources (with different species of yeast) kombucha fermentation were studied, they will kombucha at room temperature (20 ~ 22oC) culture of the 60d, the detection thereof the fermentation process. The

results of different yeasts on the use of different rates of glucose and fructose, metabolic process is not the same. Yeast on the use of sugar has played a decisive role, and use different ways of yeast is different. They used Acetobacter Xylinumm can metabolize lactic acid, so they did not detect lactic acid (Sievers, 1995).

1.4 Kombucha component analysis

Some scholars have analyzed the components contained in Kombucha tea, since the adoption of the strains, culturing conditions, analytical methods, the focus of analysis and other aspects are different, so different data and the results are different. Kombucha contains glucuronic acid, gluconic acid, acetic acid, alcohol, lactic acid, amino acids, proteins, folic acid, lichen acid, Vc and a variety of B vitamins and so on(Wu, 2012) . Two kinds of kombucha from Taiwan be analyzed by HPLC, it showed that the Kombucha contains glycerine, acetic acid and ethanol(Liu, Hsu, 1996).

The use of HPLC / MS / PDAD (High Performance Liquid Chromatography / Mass Spectrometry / optical scan) method 19 kombucha samples from the United States and elsewhere in the world carried out chemical analysis results did not find the former almost universally accepted functional factor - glucuronidase, and the content of various vitamins rarely, but checked out a lot of their predecessors did not detectable substance. They detected from 19 kinds of kombucha 40 kinds of chemical substances. These substances enter the body how to function needs further scientific research(Roussin, 1996).

Tea itself contains an abundance of nutrients. The combination of kombucha fermented tea can produce some new nutrients. Global aspects of Kombucha nutrients have a lot of research.Kombucha beverage is detected from the: 14 kinds of alcohols, acids, 13 kinds, four kinds of esters, aldehydes two kinds, other (including ether) two

kinds. The results also show that the pre-fermentation (0 ~ 4d) is mainly produced alcohols, for the following acids (citric, ascorbic, nonahexacontanoic acid, etc.) (Zhao,2010).

Polyphenols with a variety of biological activity. Kombucha beverages to detect a variety of polyphenols, such as EGCG, GCG, EGC, ECG, EC and so on. EGCG (18%) and ECG (23%) degradation throughout the Kombucha fermentation process rare [21 - 22]. These catechins reservations nutritional properties of Kombucha has a more important role (Jiang,Jayabalan,2007).

Kombucha beverage contains vitamins (B1, B6, B12, C) and minerals (zinc, copper, iron, manganese, nickel and cobalt). In the study Kombucha beverage antioxidant also found that: with the increase of fermentation time, ascorbic acid (Vc) content increased. This is illustrated in the fermentation process, kombucha metabolites produced synthetic vitamin C. Many studies have confirmed that kombucha drinks contain relatively rich in nutrients (Song,2013).

Kombucha samples were analyzed by atomic absorption chromatography and thin layer chromatography kombucha minerals and soluble vitamins , the results showed that trace elements beneficial to human body zinc, copper, iron, manganese, nickel and cobalt, these elements contents are higher than tea solution which is not be fermented, the content of harmful elements are little in kombucha such as lead and chromium, also the content of harmful elements are less than tea solution. This shows that by microbial action, can increase the beneficial element while reducing harmful elements(Bauer-Petrovska, 2000).

1.5 Anti-Oxidation of kombucha fermented tea

By the fermentation of different kinds of tea from the Kombucha drink has good antioxidant activity, it is worthy of further study and potential application made by the fermentation of Kombucha drink tea waste in inhibiting excessive free radical

generation caused by disease effect. Found in kombucha fermentation process showed growth in free radical scavenging ability. The extent and vigor radical scavenging depends on the fermentation time, teas and kombucha normal flora, which in turn determine the form of metabolites (Jayabalan,2007). The study found in the pre-fermentation 5d, kombucha drinks DPPH radical scavenging hydroxyl radical and gradually increased, but after culture 5d scavenging capacity but lower (Feng,2009). By in vitro radical scavenging experiments to study the effects of Kombucha bacteria and fermentation time source for kombucha antioxidant activity. Kombucha prove different sources have different antioxidant activity, and most show a time-dependent, but it is not recommended to extend the fermentation time, because the accumulation of organic acids may affect drinkable. Total phenol content in addition to 98%, indicating that during fermentation tea red pigment may be biodegradable and release with higher antioxidant activity of small molecules (Chu,2006). To study the mechanism of the antioxidant potential of antioxidants, kombucha drinks fractionation and further identification is in progress.

Gluconic acid, acetic acid bacteria (Gluconacetobacter sp.A4) has a strong production capacity of DSL, Kombucha is the main function of strain. Research on traditional kombucha fermented tea Kombucha fermented tea and modified (only acetic acid bacteria to ferment gluconate) drop steroids and antioxidant activity, found improved Kombucha fermented tea in oxidation and reduction effect as conventional steroids Kombucha fermented tea is similar, even stronger than it (Yang,2009).

1.6 Mechanism of Action

Mechanism of action related to kombucha beneficial to human health had not yet been completely understood, but experts and scholars at home and abroad through research, from the nature and composition of the starting preliminary kombucha explained its

mechanism of action.

1.6.1 Cleaning up the stomach to help digestion, inhibit harmful bacteria, prevention of gastrointestinal diseases, prevent colon cancer (Wu,2012). Kombucha can reduce the pH of the stomach, and the acidification of the upper gastrointestinal tract health is very good, kombucha take effect of antagonism on gastrointestinal tract of harmful bacteria, the harmful bacteria, such as Helicobacter pylori is closely related to gastritis,ulcers and stomach cancer, but also kombucha can prevent colon cancer. The study of intestinal bacteria glucosidase and glucuronidase characteristics found two enzymes are enhanced with increasing pH, when enzyme at pH7 the activity is more effective 5 to 10 times enzyme at pH6, they think high pH environment can induce the activity of these two enzymes, leading to colon cancer (Kim,1994).

1.6.2Drinking Kombucha can maintain normal blood pH. Medical studies have shown that cancer patients blood pH value of 7.56, while the body's normal blood pH value of 7.5 or less. Serological blood test showed that Kombucha beverage can significantly maintain normal blood pH, this study found that kombucha prevention and treatment of early cancer causes are important (Kim,1994).

1.6.3 The main component of Kombucha - acetic acid, is very useful for health matter. Modern medicine has proved that acetic acid has to eliminate fatigue, increase appetite, help digestion, prevention and treatment of hypertension and arteriosclerosis, Also have some anti-tumor, anti-obesity, so that calcium is easily absorbed by the body to maintain flexibility as well as the role of skeletal muscle rejuvenation run (Cui,1994).

1.6.4Many people believe that one function of detoxification cancer factor Kombucha is glucuronide, which is one of the most important human liver detoxification of substance. Endogenous toxic substances glucuronide with toxins or produce human metabolism of exogenous combined into a water-soluble glucuronide,

and excreted, so that the body may cause a variety of diseases or toxins disease has occurred was promptly removed, play prevention and cure of good results (Wu,2012).

1.6.5Glucose acid 1,4-lactone exist in Kombucha make heparin, hyaluronic acid, mucopolysaccharides and glucuronide the destruction of greatly reduced, helping the body to more effectively release toxins, anti-cancer, and relieve joint Other discomfort inflammation, gout, asthma and other discomfort caused by human tissue associated function decline (Wu, 2003).

1.6.6 Kombucha glucose acid can be combined to form a water-soluble heavy metal complexes, helping to flush out harmful heavy metal elements. Studies have shown that the new kombucha drinkers increased content of heavy metals in urine (Bauer-Petrovska, 2000).

1.6.7 Kombucha contains many tea extract, the extract of these health effects can not be ignored. According to the study of ancient and modern tea health functions up to more than 30, one of the major anti-clotting, lowering blood pressure, prevention of coronary heart disease, diuretic, aid digestion, detoxification and anti-bacterial, anti- cancer, anti-radiation (Ruan,1994). Tea polyphenols have function that can prevent mutation, is a natural antioxidant, can delay human aging; tea chlorogenic acid, catechin, Vitamin C, Vitamin E and others have to protect the skin's function.

There are many health benefits of kombucha known or unknown composition, it is healthy for some reason is not one substance alone but the result of interaction of a variety of beneficial ingredients, its role in human health is multifaceted.

1.7 Production Method for different kinds of tea

1.7.1 Green tea

Green tea is the most widely studied of tea. Just picked from the tea leaves or buds are called Cha Qing. Compared with other plant leaves, Cha Qing rich in polyphenols and caffeine. And like other green plants the same, including a large number of

chlorophyll and polyphenol oxidase. In Cha Qing, the chlorophyll color account for a dominant. But chlorophyll is unstable, it is easy be decomposed and lose the green color, after the other color of the pigment will be reflected. On the other hand, tea polyphenols are not stable, it is easy to be oxidized by the plant's own polyphenol oxidase, and aggregated into theaflavins, tea and red pigment and other substances. Green tea is to keep the core of the green, the key is picked after Cha Qing, as soon as possible inactivation of polyphenol oxidase, thus avoiding oxidation of polyphenols. By high temperature treatment, the activity of polyphenol oxidase lost, it will not be oxidized polyphenols, achieve this goal by keep green color. Then fast drying, Cha Qing state was preserved down to give green tea (Yun ,2014).

1.7.2 Yellow tea

After fixing, if not timely and dried, then the decomposition of chlorophyll may also lose the green, so that leaves the carotenoids lutein and color. They are more stable than chlorophyll, So the tea appear yellow color. Make a refinement of the process so that yellow carotenoids and lutein will be fully demonstrated, you will get pale-yellow tea , and tea solution will be pale yellow after tea leaves be immersed. Can be seen the leaves and solutions are yellow color, then the yellow tea is produced. In the production of yellow tea, this delicate process is called "stuffy yellow", is the use of heat Cha Qing fixing after that Cha Qing yellowing. In green tea was preserved at Cha Qing state, but also the grass flavor was preserved. And after stuffy yellow, this the grass flavor disappeared (Wang ,2015).

1.7.3 White tea

Core technology of white tea is withering. Cha Qing placed under natural conditions, which made the polyphenol oxidase, peroxidase and other enzymes exerted their effects. In the strictly regulate the production of white tea, withering time up to 60 hours. Sunny weather and mild north wind dried to ensure proper temperature and

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写作形式与撰写规范及格式模板

广州大学成人高等教育毕业论文(设计) 撰写规范及格式模板 一、毕业论文(设计)写作形式 1.毕业论文(设计)写作形式:毕业论文、读书报告、调查报告、案例分析、分析报告、方案策划、教学设计、毕业设计等 2.申报学士学位的学生必须撰写毕业论文或毕业设计; 3.不申报学位的学生按专业特点可选择以上毕业论文(设计)的各种写作形式。 二、毕业论文(设计)撰写内容要求 毕业论文包括:标题,摘要和关键词,目录,论文正文,参考文献,附录等;毕业设计包括工程图或作品,设计(计算)说明书。 毕业设计(计算)说明书是对毕业设计进行解释与说明的书面材料,在写法上应注意与论文的区别点是:1.前言包括设计的目的和意义,设计项目情况简介或社会调查情况概述,设计思想或设计定位;2.正文包括方案的论证,方案的设计或主要参数的计算两大部分。 毕业论文(设计)主要部分内容: 1. 标题:应简明突出、有概括性。题目字数要适当,不宜超过20个字,如果有些细节必须放进标题,可以分成主标题和副标题。 2.摘要和关键词:摘要应反映论文的精华,概括地阐述论文(设计)研究的基本观点、主要研究内容、研究方法、取得的成果和结论。要求英语专业必须有中、英文摘要,其他各专业只需中文摘要。中文摘要在300字左右,外文摘要以300个左右实词为宜。关键词是供检索用的主题词条,应采用能覆盖论文主要内容的词条。关键词一般列3~5个,按词条的概念的外延层次排列(外延大的排在前面)。 3.目录:目录中的标题应与正文中的标题一致,要求至少有二个层次,注明页码。附录也应依次列入目录。

4.正文:包括前言、正文主体与结论,一般不少于5000字。 (1)前言:前言应说明本课题的研究目的和意义、研究范围、研究方法及要达到的要求;简述本课题在国内外的研究或发展概况及存在的主要问题;说明本课题应解决的主要问题。 (2)正文主体:正文主体是对毕业论文内容的详细表述,其内容一般包括:a问题的提出,研究工作的基本前提、假设和条件。 b模型的建立,实验方案的拟定;基本概念和理论基础;设计或计算的主要方法、内容;实验方法、内容及其分析;理论论证,理论在课题中的应用; c课题得出的结果以及对结果的讨论等。 学生可根据毕业论文课题的性质,涉及上述部分内容。 (3)结论:结论是对整个研究工作进行归纳和综合而得出的总结,对所得结果与已有结果的比较,提出课题尚存在的问题和进一步开展研究的见解与建议。结论要写得概括、简短。 5. 注释:毕业论文(设计)中有个别名词或情况需要解释时,可加注说明,注释一律用篇末注(将全部注文集中在文章末尾)。按文中出现的先后顺序统一用①②自然编号,文中的序码格式为上标。 6.参考文献:参考文献是毕业论文(设计)不可缺少的组成部分,它反映了毕业论文(设计)的取材来源,材料的广博程度和可靠程度,也是作者对他人知识成果的承认和尊重,一份完整的参考文献是向读者提供一份有价值的信息资料。 参考文献只限于列出作者亲自阅读过的、最主要的、发表在公开出版物上的文献。参考文献不少于5篇,并按文中出现的顺序列出直接引用的主要参考文献。 7.附录:对于一些不宜放在正文中,但又具有参考价值的内容(例如公式的推导、编写的计算机程序、实验数据、问卷调查表等)可以编入毕业论文(设计)的附录中。 8.致谢:简述自己通过本论文研究的体会,并对指导教师以及在论文撰写过程中直接给予帮助的有关人员表示谢意。 三、毕业论文(设计)计算机打印排版格式 1.纸张和页面要求 毕业论文(设计说明书)文稿纸统一用A4 纸打印,上下页边距为2.54cm、左边距为3.0cm、右边距为2.6cm、正文行距为固定值23磅。单面打印。 2.体例 标题:论文正文按章、节、条、款、项分级,在不同级的章、节、条、款、项阿拉伯数字编号之间用点“.”(半角实心下圆点)相隔,最末级编号之后不加点。排版格式见下表。

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