ABSTRACT

The B vitamins are essential in the human diet. Many food-grade bacteria such as lactic acid bacteria produce excess of B vitamins such as folate (vitamin B9) and cobalamin (vitamin B12). This unique property of bacteria offers the possibility to fortify raw food materials such as ugba with B vitamins by natural means. This study was aimed at determining the folate and cobalamin levels in cultures of lactic acid bacteria from Pentaclethra macrophylla seeds. Four LAB species were isolated from Pentaclethra macrophylla of which were Lactobacillus fermentum, Lactobacillus plantarum, Lactococcus lactis and Leuconostoc mesenteroides.based on morphological, biochemical and physiological characteristics. The levels of folate and cobalamin in the cultures of LAB from Pentaclethra macrophylla were analyzed using High Performance Liquid Chromatography (HPLC) method. This showed the concentration of both B vitamins in LAB fermented Pentaclethra macrophylla to be 0.1125 mg/ml (folate) and 0.1154 mg/ml (cobalamin) and that of unfermented Pentaclethra macrophylla to be 0.3112 mg/ml (folate) and 0.0624 mg/ml. From this study, it was seen that the isolated LAB used in fermentation of Pentaclethra macrophylla were able to increase the folate and cobalamin levels by 72.26% and 45.93% respectively. Therefore, it has been shown that increased folate and cobalamin levels in fermented Pentaclethra macrophylla (ugba) are possible through judicious selection of the microbial species and cultivation conditions. 

TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgment iv

Table of contents v

List of tables viii

List of figures ix

Abstract x

CHAPTER ONE

INTRODUCTION 1

1.1 Aim and Objectives of the Study 2

1.1.1 Aim 2

1.1.2 Objectives 2

CHAPTER TWO

LITERATURE REVIEW 3

2.1 Fermentation 3

2.2 Ugba 4

2.2.1 Nature of the Plant and the Seeds 5

2.2.2 Chemical Composition of Seeds 5

2.3 Preparation of Ugba 7

2.4 Microorganisms Involved in the Fermentation of African Oil Bean Seed 9

2.5 Nutritional Value of Ugba 10

2.6 Lactic Acid Bacteria 11

2.6.1 Characteristics of Lab 12

2.6.2 Use of Lactic Acid Bacteria in Food 13

2.7 B-Vitamins 14

2.8 Folate or Folic Acid 15

2.8.1 Production of Folate by Lactobacillus 16

2.9 Vitamin B12 17

CHAPTER THREE

MATERIALS AND METHOD 19

3.1 Study Area 19

3.2 Collection of Samples 19

3.3 Culture Media Preparation and Sterilization 19

3.4 Preparation of Homogenate Ugba Samples 20

3.5 Inoculation of Samples 20

3.6 Colony Count of Isolates 20

3.7 Characterization and Identification of Isolates 21

3.8 Identification of Lactic Acid Bacteria 21

3.8.1 Biochemical Characterization 21

3.8.1.1 Gram Staining 21

3.8.1.2 Catalase Test 22

3.8.1.3 Spore Stain Test 22

3.8.1.4 Citrate Test 23

3.8.1.5 Carbohydrate Fermentation Test 24

3.8.1.6 Haemolytic Activity    24

3.9 Physiological Characterization of Lactic Acid Bacteria 24

3.9.1 Growth at Different Temperatures 24

3.9.2 Salt Tolerance Test 24

3.9.3 Growth at Different pH 25

3.10 Laboratory Fermentation of Ugba Using Lab Isolates for Vitamin Production 25

3.11 Evaluation of the Folate and Cobalamin Levels of the Isolated LAB 25

CHAPTER FOUR

RESULTS 27

CHAPTER FIVE

DISCUSSION, CONCLUSION AND RECOMMENDATION 36

5.1 Discussion 36

5.2 Conclusion and Recommendation 39

References 40

Appendix I 46

Appendix II 47

Appendix III 48

Appendix IV 49