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GELATINASE PRODUCTION POTENTIAL OF BACTERIA ISOLATED FROM ATIMBO AND KWA RIVER SEDIMENTS
ABSTRACT
The industrial importance of gelatinase cannot be overemphasized as more jaw dropping discoveries have uncovered the various undisputable applications of the enzyme in several aspects of science. In this study, gelatinase producing bacteria were probed from sediment samples of Kwa River and Atimbo River, Cross River State. Primary screening uncovered 3 isolates (B5, A7 and B6) that could elaborate gelatinase activity. This exposed the quantity of gelatinolytic bacteria to be 5.4% of the entire sediment sample population. The isolates that showed positive gelatinolytic activity were then quantitatively screened using the Bradford method which displayed high production in 2 isolates (B6 and B5). The isolate that showed maximum gelatinolytic activity was then centrifuged and evaluated to determine the extracellular production of the enzyme using cell free supernatants. The cell free supernatants of positive isolates were used as crude gelatinase. Determination of carbon source required by the bacteria for optimal production of the gelatinase showed high rates of enzyme activity in sucrose which was selected. The successful organism was identified to be of Bacillus species using identification tests.
TABLE OF CONTENTS
Title page i
Certification ii
Dedication iii
Acknowledgements iv
Abstract v
Table of Contents vi
List of Figures vii
List of Plates viii
List of Tables ix
CHAPTER ONE: INTRODUCTION 1
Aim and objectives of study 3
Justification of study 3
CHAPTER TWO: LITERATURE REVIEW 4
Gelatins 4
Sources of gelatin 7
Production of gelatin 9
Gelatin structure 12
Mechanism of gelatin gelation 14
Properties of gelatin 15
Industrial applications of gelatin 17
Proteases 21
Microbial proteases 22
Applications of microbial proteases 24
Matrix Metalloproteases (MMPs) 27
Gelatinase 29
Types of gelatinases 30
Structure of gelatinases 31
Activation of gelatinases 32
Initiation of gelatinases 33
CHAPTER THREE: MATERIALS AND METHODS 34
Materials 34
Methods 34
Sampling 34
Sample collection 34
Sample preparation 35
Microbial analysis 35
Enumeration 36
Isolation 36
Primary screening for gelatinase-liquefying bacteria 36
Secondary screening for gelatinase production in submerged culture 37
Evaluation of carbon sources for optimal gelatinase production 39
Characterization and identification of selected
gelatinase-producing bacterium 41
CHAPTER FOUR: RESULTS 42
Microbial analysis 42
Primary screening for gelatinase-liquefying bacteria 42
Secondary screening for gelatinase production in submerged culture 43
Evaluation of carbon sources for optimal gelatinase production 43
Characterization and identification of selected
gelatinase-producing bacterium 44
CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATION 49
Discussion 49
Conclusion 50
Recommendation 51
References
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