Enzymatic Potentials Of Lactic Acid Bacteria And Saccharomyces Cerevisiae From Fresh And Soured Palm Wine

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