SCREENING FOR LIPASE-PRODUCING BACTERIA FROM TRADITIONAL FERMENTED FOODS (TFFs)

ABSTRACT

The study is aimed at isolating and identifying bacterial organisms producing lipase from some traditional fermented foods in Umuahia main market, Abia State. Traditional fermented foods such as Ogi, Ugba and Ogiri were used, medias used were MacConkey agar, nutrient agar (modified) blood agar and mannitol salt agar. One gram of each samples were weighed and homogenized in 5mls of sterile normal saline and serially diluted up to the fifth (105) dilution and 0.02ml cultured using the streak plate method on the media prepared and incubated for 24hrs at 370C and identification of lipase producing isolate was done based on their morphology, gram staining and biochemical test were carried out. Organisms such as Proteus spp, Staphylococcus aureus, Micrococcus spp, Pseudomonas spp and Bacillus spp were identified as isolates producing lipase by showing a halo or zone of clearance around the colonies in olive oil incorporated media. Ugba was found to habour the highest number of lipase producing bacteria followed by ogiri and ogi containing 7,5 and 3 organisms with positive lipolytic activities respectively. In conclusion traditional fermented food should be handled, packaged and transported properly to avoid the chance of contamination and lipase production is effectively useful and should be well understood.  

TABLE OF CONTENTS

Certification ﾿ i

Dedication ﾿ ii

Acknowledgement ﾿ iii

Table of Contents ﾿ iv

List of Tables ﾿ vii

List of Figures ﾿ viii

Abstract ﾿ ix

CHAPTER ONE

1.0 ﾿ INTRODUCTION ﾿ 1

1.1 ﾿ Background of the Study ﾿ 3

1.2 ﾿ Aim of Study ﾿ 3

1.3 ﾿ Objectives of the Study ﾿ 3

1.4 ﾿ Limitations ﾿ 3

CHAPTER TWO

2.0 ﾿ LITERATURE REVIEW ﾿ 4

2.1 ﾿ Traditional Food Fermentation ﾿ 4

2.2 ﾿ Major Traditionally Fermented Foods in Nigeria ﾿ 5

2.2.1 ﾿ Ogi ﾿ 5

2.2.2 ﾿ Ogiri ﾿ 5

2.2.3 ﾿ Ugba ﾿ 6

2.2.4 ﾿ Afiyo (Okpehe) ﾿ 6

2.2.5 ﾿ Dadawa/Iru ﾿ 7

2.3 ﾿ Nutritional Benefits and Health Benefits of Traditional Fermented Foods ﾿ 8

2.3.1 ﾿ Nutritional Benefits ﾿ 8

2.3.2 ﾿ Health Benefits ﾿ 9

2.4 ﾿ Lipases ﾿ 13

2.4.1 ﾿ Sources of Lipases ﾿ 13

2.4.2 ﾿ Properties and Reactions of Lipases ﾿ 13

2.5 ﾿ Factors affecting Microbial Lipase Production ﾿ 14

2.5.1 ﾿ Effect of Nutritional Factors ﾿ 15

2.5.2 ﾿ Effects of Physical Factors ﾿ 17

2.6 ﾿ Application of Lipases for Industrial Processes ﾿ 20

CHAPTER THREE

3.0 ﾿ MATERIALS AND METHODS ﾿ 22

3.1 ﾿ Study Area ﾿ 22

3.2 ﾿ Materials and Reagents ﾿ 22

3.3 ﾿ Media Preparation ﾿ 22

3.4 ﾿ Sterility ﾿ 23

3.5 ﾿ Sample Collection ﾿ 23

3.6 ﾿ Sample Preparation Inoculation and Isolation ﾿ 23

3.7 ﾿ Lipolytic Activity Assay ﾿ 23

3.8 ﾿ Identification Lipase producing Isolates ﾿ 24

3.8.1 ﾿ Gram Staining ﾿ 24

3.8.2 ﾿ Catalase Test ﾿ 24

3.8.3 ﾿ Coagulase Test ﾿ 25

3.8.4 ﾿ Citrate Test ﾿ 25

3.8.5 ﾿ Motility, Indole, Urease Test (MIU) ﾿ 25

3.8.6 ﾿ Triple Sugar Iron Test ﾿ 26

3.8.7 ﾿ Oxidase Test ﾿ 26

CHAPTER FOUR

4.0 ﾿ RESULTS ﾿ 27

CHAPTER FIVE

5.0 ﾿ DISCUSSION ﾿ 37

5.1 ﾿ Recommendation and Conclusion ﾿ 38

References 

LIST OF TABLES

Table ﾿ Title ﾿ Page

1: ﾿ TFFs Sampled and Number of Bacterial Organisms with Lipolytic 

﾿ Activity Isolated ﾿ 28

2: ﾿ Morphological Identification of Lipase producing Isolates ﾿ 29

3: ﾿ Biochemical Characterization Lipase producing Isolates ﾿ 30

4: ﾿ Comparative Analysis of Lipase producing Bacterial Diversity of 

﾿ sampled TFFs ﾿ 31

5: ﾿ Frequency of Occurrence of Lipase producing Isolates ﾿ 32

LIST OF FIGURES

Plate ﾿ Title ﾿ Page

1: ﾿ Samples inoculated in media ﾿

3: ﾿ Bacteria growth observed in Ogiri ﾿ 34

4: ﾿ 34

5: ﾿ Bacterial Growth Observed in Pap (Ogi) ﾿ 35

6: ﾿ ﾿ Bacterial Growth observed in Nutrient agar mixed with olive oil ﾿ 35

7: ﾿ Clear zone observed (Zone of inhibition) showing lipase production ﾿ 36