Amylase Production By Solid State Fermentation Of Agro-Industrial Waste Using Bacillus Species

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