LACTIC ACID BACTERIA MEDIATED FERMENTATION OF BEETROOT (BETA VULGARIS)

ABSTRACT

This work explores 10 days fermentation of beetroot juice using Lactic Acid Bacteria (Lactobacillus plantarum and Lactobacillus bulgaricus) which was carried out to enhance the nutritional quality and shelf life of beetroot. Beetroot was analyzed using the standard methods for proximate analysis. Carbohydrate was calculated by difference, the mineral content of beetroot was determined by the dry acid extraction method. Lactic acid bacterial load during the course of fermentation was checked by pour plate technique. Proximate analysis of beetroot showed that there was increase in nutrient composition during the course of fermentation. Result of proximate composition of beetroot (Beta vulgaris) indicates that the highest protein content was achieved with L. bulgaricus (1.87%) on the 10th day of fermentation. The fat decrease as fermentation proceeds and the highest decrease occurred in L. bulgaricus fermented juice (0.04%). The percentage ash increased slightly in both fermented juices but higher in the fermented juice inoculated with L. bulgaricus (1.19%). The moisture content decreased and the highest decrease was achieved with L. bulgaricus (84.44%). Carbohydrate content increased and was higher in the sample inoculated with L. bulgaricus (12.46%). The result in mineral content of the fermenting beetroot juice decreased significantly with L. plantarum and L. bulgaricus as fermentation time increase. The decrease in calcium was higher with L. plantarum (1.68mg/100ml). Magnesium also reduced and the decrease was higher with L. plantarum (1.12mg/100ml). Similarly, potassium and sodium reduced significantly with fermentation time and the highest decrease was achieved with L. plantarum (3.17mg/100ml and 6.92mg/100ml respectively). The decrease in phosphorus was higher in L. bulgaricus fermented juice (13.34mg/100ml). The result of Lactic acid bacterial load shows that there was an initial high increase from the zero day to the 2nd day on both fermenting juice. Then, a maximum increase was recorded for both Lactobacillus species after 4 days of fermentation whereby the mean Lactic acid bacterial loads stood at 2.08 × 107 CFU/ml and 2.56× 107 CFU/ml for L. plantarum and L. bulgaricus fermented juice respectively. From the 6th day of fermentation, a decline was recorded for both sample in which the load decreased from 2.08 × 107 CFU/ml to 1.01 × 107 CFU/ ml at the 10th day of fermentation by L. plantarum while the load decreased from 2.56 × 107 CFU/ml to 1.18 × 107 CFU/ml in L. bulgaricus fermented beetroot juice. This result reveals that the beetroot juice contain appreciable amounts of nutrients that justifies its use in treatment of different ailments and also its fermentation will make it a good functional beverage. 

TABLE OF CONTENT

Title page                                                                                                                                         i

Certification                                                                                                                                     ii

Dedication                                                                                                                                          iii

Acknowledgement                                                                                                                              iv

Table of contents                                                                                                                                         v

List of tables                                                                                                                                        vii

List of pictures                                                                                                                                   viii

Abstract                                                                                                                                                  ix

CHAPTER ONE

1.1 INTRODUCTION                                                                                                                  1-3   

1.2 Aim and Objectives                                                                                                                  3-4

CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Fermentation                                                                                                                            5-6

2.2 Purposes of fermentation                                                                                                                 6

2.3 Advantages of fermentation                                                                                                              7

2.4 Disadvantages of fermentation                                                                                                           7

2.5 Factors hampering the development of fermented food                                                                 8

2.6 Beetroot                                                                                                                                   8-9      

2.7 Health benefits of consuming beetroot                                                                                  9- 13 

2.7.1 Bioavailability                                                                                                                   13-14

2.72 Oxidative Stress                                                                                                                 14-15

2.7.3 Inflammation                                                                                                                                15

2.8 Fermentation of Beetroot juice                                                                                                      15

2.9 Lactic acid bacteria                                                                                                             16-17

CHAPTER THREE

3.0 MATERIALS AND METHOD        

3.1 Source of Materials                                                                                                                               18

3.2 Sample Preparation                                                                                                                       18

3.3 Fermentation of Beetroot Juice                                                                                            18-19

3.4 Inoculation of Beetroot Juice                                                                                                             19

3.5 Proximate Analysis                                                                                                                   19 

3.5.1 Ash Content Determination                                                                                                                20

3.5.2 Fat Content Determination                                                                                                20-21

3.5.3 Crude Protein Determination                                                                                            21-22

3.5.4 Determination of Moisture Content                                                                                              23

3.5.5 Determination of Carbohydrate                                                                                        23-24

3.6 Determination of Minerals                                                                                                         24

3.6.1 Determination of Calcium and Magnesium                                                                      24-25

3.6.2 Determination of Potassium and Sodium                                                                         25-26

3.6.3 Determination of Phosphorus                                                                                           26-27

3.7 Determination of Lactic acid bacterial Load                                                                              27

3.8 Statistical Analysis                                                                                                                      28

CHAPTER FOUR

4.0 Result                                                                                                                                         29

CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1 Discussion                                                                                                                            33-36

5.2 Conclusion                                                                                                                                     36

5.3 Recommendation                                                                                                                        37

Reference                                                                                                                                    38-46

Appendix                                                                                                                                   47-49                                                                                                                        

List of Tables

Table 1                                                                                                                                                          30

Table 2                                                                                                                                                     31

Table 3                                                                                                                                                 32

 List of Pictures

Picture 1                                                                                                                                                      50                                          

Picture 2                                                                                                                                              51

Picture 3                                                                                                                                                52

Picture 4                                                                                                                                              53

CHAPTER ONE

1.1   INTRODUCTION

Beta vulgaris (Chenopediceae) also known as garden beet or red beet in the US scientific literature and beetroot in Europe and many other countries around the world, is a herbaceous biennial or rarely perennial plant. It has a long history dating to the second millennium BC. The first cultivated forms were believed to have been domesticated in the Mediterranean, but were introduced to the Middle East, India and finally China by 850AD.

These were used as medicinal plants in Ancient Greece and medieval Europe. Their popularity declined in Europe following the introduction of spinach. Beta vulgaris is best known in its numerous cultivated varieties, the best known of which is the purple root vegetable known as the beetroot or garden beet. The “earthly” taste of some beet root cultivars comes from the presence of geosimin. Researchers have not yet answered whether it is produced by symbiotic soil living in the plant (Lu et al., 2003). Beetroot contains the bioactive agent betalains which supports healthy liver function (Vali et al., 2007). It was found that beetroot inhibits nitrosodimethylamine (NDMA) formation 96). In a study to determine the estimated Glycemic index of various foods, it was concluded that beetroot has a medium G.I of 64 (Babek et al., 2000). It is used in treatment of cardiovascular diseases (Olth et al.,2005) and blood pressure (Ahulwalia, 2008). Proximate and nutrient analysis of medicinal plants, edible fruits and vegetables plays a crucial role in assessing their nutritional significance (Pandey et al., 2006). As various medicinal plant species are also consumed as food along with their medicinal benefits, evaluating their nutritional significance can help to understand the worth of these plant species (Pandey et al., 2006). This study therefore focuses on the proximate composition, mineral analysis of Beta vulgaris root with a view to assess the nutritional potential in relation to its ethno medicinal uses.

Beetroot is the root of the sugar beet crop. The crop is botanically called Beta vulgaris. It is a measure of the plant family Chenopodiaceae and is reported to be the second important sugar crop after sugar cane. According to Brar et al., (2015), the crop is regarded as a crop of the temperate region but as currently spread to many sub-tropical countries across many continents of the world including: Africa, America, Europe. Kumar and Pathak (2013) reported that the crop is grown successfully in 57 countries of the world.

The beetroot is a root crop and the sugar is harvested from the roots. It is a major source of taste sugar (21.8%) in the world (Shrivastava 2006). Juices produced from beetroot are reported to be nutritionally high and with high health powering potentials. List of health benefits of the beetroot juice include: Lowering blood pressure, anti-cancer properties, body detoxification, immune boosting etc.

Lactobacillus are probiotics and have been successfully used in the fermentation diary and other products. They are also known to belong to GRAS microorganisms (Generally Regarded As Safe). The project is expected to free impact into the activity of the Lactobacillus as well as juice fermentative products.

Beta vulgaris, commonly known as red beet or sugar beet, is the most common variety found in Britain, Central as well as North America.

Researchers are now focusing on functional beverages, made from fruits and vegetables by means of controlled fermentation using probiotic bacteria such as

L. bulgaricus, L. plantarum, Streptococcus thermophillus, L. casei, Bifidobacterium bifidum, B. infantis, since they are abundantly present in the human digestive tract. 

Fermented functional beverages are non-alcoholic beverages, which on consumption bring about an increase in the total number of Lactic acid bacteria (LAB) in the intestinal tract, which helps in enhancing immunity against some common pathogens. Fermented vegetable and fruit beverages are also very beneficial for people with lactose intolerance.

Among many other health benefits, fermented functional beverages are also expected to help in the prevention of different kinds of cancers like colon cancer, since the LAB present in the beverage decrease the level of carcinogens and an enzyme called β glucouronidase. After a few trials in humans, it was again suggested that the anti-cancer activity of LAB could be due to a decrease in the activity of enzyme glucuronidas. However, the data for humans is not sufficient to well establish the research. Therefore, this research aimed at producing a functional beverage by fermenting Beta vulgaris by a co-culture of L. plantarum and L. bulgaricus to determine the proximate composition, Mineral analysis of the beetroot juice as well as the microbial load of the fermented beetroot juice.

1.2   AIM AND OBJECTIVES

The aim of this project work is to ferment beetroot juice using Lactobacillus species. Against the above backgrounds, this project was conceived to study the fermentation of beetroot juice using Lactobacillus species.  The objectives include specifically the following:

·         Production of juice from beetroot.

·         To use different Lactobacillus species to ferment beetroot juice.

·         Evaluation of fermentative dynamics of the fermenting juice through the assessment of the different products of fermentation.

·         Determination of proximate composition and mineral content of the fermented juice.

·         Comparison of the fermentative capacity of the different species of Lactobacillus.

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