FOLATE AND COBALAMIN LEVELS IN CULTURES OF LACTIC ACID BACTERIA FROM PENTHACLETHRA MACROPHYLLA (UGBA) SEEDS

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