ABSTRACT

The enzymatic potentials of lactic acid bacteria and Saccharomyces cerevisiae  from fresh and soured palm wine samples were determined from 50 different palm win samples collected from Ariam, Umudike and Umunneochi in Abia state. Lactobacillus planetarium and Leuconostic  Mesenteroides were the dominated  lactic bacteria and Saccharomyces cereuisiae and Saccharomyces uvarum were the dominated yeast species isolated from the palm wine sample the lactic acid bacteria and saccharomyces cerevisiae isolated were screened for amylase, cellulase, protease, pectinase and xylanase  production   using well plate method  in order to determine their ability to produce these enzymes. Among 88 LAB isolated, 20 were able to exhibit amylase production,  15 produced protease, 10 produced pectinase, 7 produced cellulose and 5 produced xylanase with different zone of inhibition . The isolate LAB 35 was able to produce all the enzymes tested and it was selected and identified as Lactobacillus plantarium . In other hand, among the 50 S. cerevisiae  isolated, 10 were able to exhibit amylase production, 5 produced cellulose, 8 produced protease, 7 produced pectinase and 2 produced xylanase with different halo zones. Amonge the isolates that were able to produce these enzymes S.cerevisiae 25 has the potential to produce all the enzymes tested (amylase, protease, cellulose, pectinase and xylanase). 

TABLE OF CONTENTS 

Cover page 

Title page i

Declaration ii

Certification iii

Dedication iv

Acknowledgement v

Table of contents vi

List of table xi

List of figure xii

Abstract xiii

CHAPTER ONE : INTRODUCTION 

1.1 Aims 2

1.2 Objectives 2

CHAPTER TWO: LITERATURE REVIEW 

2.1 Palm Wine 3

2.1.1 Palm Wine Production 3

2.1.2 Biochemical Constituents 4

2.1.3 Microbial Communities in Palm Wine 5

2.1.4 Benefits of Palm Wine 6

2.2 Lactic Acid Bacteria 6

2.2.1 Condition that Favors the Growth of Lactic Acid Bacteria. 7

2.2.2 The activities of Lactic Acid Bacteria in Palm Wine 8

2.3 Yeast. 9

2.3.1 Conditions Necessary for the Growth of Yeast 9

2.3.2 Uses of Yeast 11

2.3.2.1 Brewing 11

2.3.2.2 Alcoholic Beverages 11

2.3.2.3 Beer Production 11

2.3.2.4 Production of wine 12

2.3.2.5 Baking 12

2.3.2.6 Food Spoilage 12

2.3.2.7 Bioremediation 13

2.3.3 Saccharomyces cerevisiae 13

2.3.3.1 Factors that Affects the Growth of Saccharomyces cerevisiae. 14

2.3.3.2 The Activities of Saccharomyces cerevisiae in Palm Wine 14

2.4 Enzyme 15

2.4.1 Types of Amylase 15

2.4.1.1 α-Amylase 15

2.4.1.2 β – Amylase 15

2.4.1.3. γ – Amylase 16

2.4.2 Sources of amylase 16

2.4.3 Microorganisms capable of producing amylase; 16

2.4.4 Industrial Applications of α-Amylase 17

2.4.4.1 Bakery Industry 17

2.4.4.2 Detergent Industry 17

2.4.4.3 Desizing of Textiles 18

2.4.4.4 Paper Industry 18

2.4.4.5 Fuel Alcohol Production 18

2.5 Xylanase 19

2.5.1 Application of xylanase 19

2.5.2 Xylanase sources 20

2.6 Cellulose 20

2.6.1 Microorganisms having cellulolytic abilities. 21

2.6.2 Application of Cellulases in Various Industries 21

2.6.2.1 Pulp and Paper Industry 21

2.6.2.2 Textile Industry 21

2.6.2.3 Bioethanol Industry 22

2.6.2.4 Wine and Brewery Industry 22

2.6.2.5 Food Processing Industry 22

2.6.2.6 Animal Feed Industry 22

2.6.2.7 Agricultural Industries 23

2.6.2.8 Detergent Industry 23

2.6.2.9 Waste Management 23

CHAPTER THREE: MATERIALS AND METHOD

3.1 Sources and collection of samples 24

3.2    Sterilization of Materials 24

3.3 Normal Saline Preparation 24

3.4        Media Preparation 24

3.5 Isolation of yeasts (saccharomyces) 24

3.6 Identification of yeast (saccharomyces) 24

3.7 Isolation of Lactic Acid Bacteria 25

3.8 Sub-culturing 25

3.9 Characterization and Identification of bacterial isolates 26

3.9.1 Gram staining techniques 26

3.9.2 Motility test 26

3.9.3 Catalase test: 26

3.9.4 Coagulase test 26

 3.9.5 Methyl red test: 27

3.9.6 Voges-proskaeur test: 27

3.9.7 Indole test: 27

3.9.8 Citrate test: 28

3.9.9 Oxidase test: 28

3.10 Sugar utilization test 28

3.11 Qualitative Screening for Extracellular Enzyme Producing  Isolate by plate assay 28

3.11.1 Production of Xylanase enzyme 28

3.11.2 Production of Amylases enzyme 28

3.11.3 Production of Cellulases enzyme 29

3.11.4 Screening of Isolates for the Pectinase Activity 29

3.11.5 Screening of Proteolytic Activity 30

CHAPTER FOUR: RESULT

4.1 Discussion                                                40

CHAPTER FIVE: CONCLUSION

5.1 Conclusion 42

References 43