ENUMERATION OF LACTIC ACID BACTERIA FROM FERMENTED AFRICAN OIL BEAN (PENTACLETHRA MACROPHYLLA BENTH) SEEDS FOR NIACIN AND PANTOTHENIC ACID PRODUCTION

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