ABSTRACT

This study evaluated amylase production by Bacillus species employing the solid state fermentation method, agro-industrial waste were used as the substrate. Five species were tested for amylase production abilities. Bacillus subtilis showed the highest amylase production ability after incubation. Corn chaff gave maximum enzyme production (3.25U/ml) at 300C, while the least enzyme was recorded on groundnut husk (2.35U/ml) at 250C Potato peel had maximum enzyme production by Bacillus subtitlis (3.05U/ml) at ph 7.0,while the least enzyme production was from groundnut husk (2.84U/ml) at pH 4.0.Thus there was an increase in enzyme production with corresponding increase in substrate concentration. The results obtained in this study support the suitability of using agro-industrial wastes as solid for high production of amylase, and is a means of solving pollution problems, thus making solid state fermentation (SSF) an attractive method. 

TABLE OF CONTENTS

Title Page   i

Certification iii

Dedication iv

Acknowledgement v

Table of Contents vi

List of Tables vii

Abstract viii

CHAPTER ONE

1.0 Introduction 1

1.1 Aim and Objectives 4

CHAPTER TWO

2.0 Literature Review 5

2.1 Solid-State Fermentation 5

2.2 Microorganisms Associated with Solid State Fermentation 7

2.3 Substrates for Solid State Fermentation 8

2.3.1 Lignocellulose 9

2.3.2 Pectins 10

2.3.3 Lignin 10

2.3.4 Starch 10

2.4 Environmental Factors Affecting Solid State Fermentation 11

2.4.1 Moisture Content and Water Activity (Aw) 11

2.4.2 Temperature and Heat Transfer 12

2.4.3 Control of pH and Risks of Contamination 13

2.4.4 Oxygen Uptake 13

2.5 Optimization of Agro-residues for Αlpha-Amylase Production by Bacillus subtilis and its Application in Detergent Industry 14

2.5.1 Optimization of Agro-Residues for Αlpha-Amylase Production 14

2.5.2 Optimization of Process Parameters 15

2.5.3 Amylase Assay 16

2.5.4 Partial Purification of Αlpha-Amylase 16

2.6 Amylase Production by Solid-State Fermentation of Agro-Industrial Wastes Using Bacillus spp 17

2.7 Bio-processing of Food Industrial Waste for Amylase Production by Solid State Fermentation 18

2.8 Solid Substrate Fermentation Using Agro Industrial Waste for Amylase Production by Bacillus Licheniformis 20

2.8.1 Agro-Waste as Substrates for Amylase Production 20

2.8.2 Fermentation Medium 21

CHAPTER THREE

3.1 Materials and Methods 22

3.2 Collection of Substrate 22

3.3 Test Bacterial Used 22

3.4 Screening of Test Bacterial 22

3.5 Solid State Fermentation Technique 22

3.5.1 Enzyme extraction 23

3.6 Amylase Enzyme Assay 23

3.7 Optimization of Fermentation Parameters for Amylase Activity 23

3.7.1 Effect of Incubation Period on Amylase Activity 24

3.7.2 Effect of Incubation Temperature on Amylase Activity 24

3.7.3 Effect of pH of the medium on amylase Activity 24

3.7.4 Effect of Substrate Concentration on Amylase Activity 24

3.8 Statistical Analysis 24

CHAPTER FOUR

4.0 Results 25

CHAPTER FIVE

5.0 Discussion and Conclusion 31

5.1 Discussion 31

5.2 Conclusion 35

References