ABSTRACT

The enumeration of Lactic Acid Bacteria (LAB) from fermented African Oil Bean  (Pentaclethra macrophylla Benth) Seeds for niacin and pantothenic acid production was studied. Locally produced samples of Pentaclethra macrophylla were procured from Ubani main market, Umuahia, Abia State. The samples were plated on M17 agar, MRS (de Man Rogosa and Sharp) agar and on Nutrient Agar. All the media used for the isolation of these organisms were prepared aseptically according to the manufacturer’s specification. Morphological and biochemical characteristics and sugar fermentation pattern were employed to identify lactic acid bacteria. Sixteen (16) lactic acid bacterial isolates which were found to be Gram positive, non-spore formers and catalase negative were isolated. The identified organisms include Lactobacillus plantarum, Streptococcus lactis, Lactobacillus fermentum, Pediococcus spp., and Leuconostoc mesenteroides. The organisms isolated were able to grow at temperature of 45oC but varied at 15oC. Majority of the isolates survived at a salt concentration of 4 to 6%. The High Performance Liquid Chromatography (HPLC) analysis results revealed that the concentration of vitamins (niacin and pantothenic acid) increased in the fermented samples. Niacin increased from 0.1367mg/ml in the unfermented Pentaclethra macrophylla sample to 0.300mg/ml in the fermented Pentaclethra macrophylla (Ugba) sample, while pantothenic acid increased from 0.5877mg/ml in the unfermented Pentaclethra macrophylla sample to 0.8843mg/ml in the fermented Pentaclethra macrophylla (Ugba) sample. This study has shown the Niacin and Pantothenic acid production potential of lactic acid bacteria from fermented African Oil bean seeds. Hence, vitamin-producing LAB should be employed in the production of vitamin rich fermented foods and drugs in other to enrich the food matrices and improve human health.

TABLE OF CONTENTS

Title page                                                                                                                              i

Certification                                                                                                                          ii

Dedication                                                                                                                            iii

Acknowledgements                                                                                                              iv

Table of contents                                                                                                                  v

List of Tables                                                                                                                        viii

List of Figures ix

List of Plates x

List of Appendices xi

Abstract                                                                                                                                  xii

CHAPTER 1

1.1 Introduction 1

1.2 Aim and objectives 3

CHAPTER 2: LITERATURE REVIEW

2.1 African Oil Bean Seeds 4

2.1.1 Nature of the plant and the seeds 5

2.1.2 Chemical composition of African oil bean seeds 6

2.1.3 Microorganisms involved in the fermentation of African oil bean seeds 7

2.1.4 Changes that occur during Ugba fermentation 8

2.1.5 Optimization of ugba fermentation 10

2.2 Vitamins 11

2.2.2 Niacin 13

2.2.2.1 Biosynthesis of Niacin 14

2.2.3 Pantothenic Acid 17

2.3 Lactic Acid Bacteria 18

2.3.1 Fermentation by LAB 19

2.3.2 Microorganisms involved in the lactic acid fermentation 19

2.4 Factors Affecting Fermentation Process 21

2.5 Effects of Fermentation 25

2.5.1 Flavour Enhancement 25

2.5.2 Nutritional Quality 25

2.5.3 Preservative Properties 26

2.5.4 Detoxification 26

2.5.5 Antibiotic Activities 27

2.1.3 Traditionally Fermented Foods in Nigeria 28

CHAPTER 3: MATERIALS AND METHODS

3.1 Source of Raw Material 30

3.2 Materials 30

3.3 Preparation of homogenate Ugba samples 30

3.4 Inoculation of samples 31

3.5 Colony count of isolates 31

3.6 Characterization and identification of isolates 31

3.6.1 Isolation and identification of lactic acid bacteria 32

3.6.2 Phenotypic characterization 32

3.7 Biochemical characterization 32

3.7.1 Gram staining 32

3.7.2 Catalase test 33

3.7.3 Spore Stain Test 33

3.7.4 Citrate Test 33

3.7.5 Amylase test 34

3.8 Characterization of Lactic Acid Bacteria 34

3.8.1 Growth at different temperatures 34

3.8.2 Growth at different pH 34

3.8.3 Salt tolerance test 35

3.8.4 Gas production from glucose 35

3.8.5 Carbohydrate fermentation test 36

3.9 In vitro/ Laboratory Fermentation of Ugba Using LAB 36

      Isolates for Vitamin Production

3.10 HPLC analysis 37

CHAPTER 4

Results 38

CHAPTER 5: DISCUSSION, CONCLUSION AND RECOMMENDATIONS

5.1 Discussion 47

5.2 Conclusion 50

5.3 Recommendation 50

REFERENCES 51

APPENDICES 58