PRODUCTION OF AMYLASE FROM FERMENTED UGBA USING Micrococcus species

ABSTRACT

Ugba is a fermented product gotten from the fermentation of African oil bean seeds (Pentaclethra macrophylla Benth). It is an important source of carbohydrate, protein and lipids. Microorganisms such as Micrococus roseus, Micrococus luteus, Leuconostoc mesenteroides, Staphylococcus epidermidis, Bacillus substilis, Bacillus megatarium, and Bacillus lichenifornis are predominant in its fermentation process where they secrete enzymes that break down the biomolecules of ugba. Micrococcus was aseptically isolated from ugba samples. 80% of the five different samples screened contain Micrococcus species. Iodine starch test colorimetric method was employed in the determination of amylase production by the isolate. Micrococcus was aseptically inoculated in a soluble starch solution and incubated for 48hours at different temperature and pH range, a test tube containing no Micrococcus was used as a control. After incubation, Micrococcus hydrolysed the starch by producing enzyme, the spectrophotometric absorbance of the blue black coloration was read; the reduction in starch activity as shown by the reduced absorbance relative to the control measured the enzyme activity signifying utilization and reduction in the starch incorporated in the media. Optimum production of amylase was seen at 600C and pH value of 6. Temperature and pH had significant effect on the rate of enzyme “amylase” production by Micrococcus. The ability of the Micrococcus isolated in this study to break down starch shows its role in amylase production which breaks down the starch molecules in Ugba and supports the use of microorganisms as starter cultures in fermentation, breaking down of complex organic molecules and in enzyme production.

TABLE OF CONTENTS

Title page                                                                                                                           i

Certification                                                                                                            ﾿        ii

Dedication                                                                                                                         iii ﾿

Acknowledgements ﾿        iv

Table of Contents ﾿         v

List of Tables                                                                                                                     viii

List of Figures ﾿         ix ﾿

Abstract ﾿         x

CHAPTER ONE 

1.1 INTRODUCTION ﾿                                             1

1.2 Aims and Objectives                                                                                                     3

CHAPTER TWO 

2.0 LITERATURE REVIEW ﾿         4

2.1 Amylase (History and Discovery) ﾿         4

2.1.1 Uses and Applications of Amylases                                                                          5

2.2 Microorganisms and Amylases ﾿         6

2.3 Production of Amylases ﾿         8

2.4 Fermented foods                                                                                                          10

2.5 Method of production of Ugba                                                                                   11

2.6 Microorganisms involved in Ugba Fermentation ﾿        12

2.7 Micrococcus species in Ugba ﾿        13

2.7.1 Industrial uses of Micrococcus ﾿        13

CHAPTER THREE 

3.0 MATERIALS AND METHODS                                                                                   15

3.1 Materials and equipment                                                                                               15         

3.1.1 Source of Materials (Sample collection) ﾿           15

3.2 Samples/Media Preparation ﾿                       15

3.3 ﾿ Isolation of Micrococcus                                                                                                16  

3.4 Characterization of isolate for identification of Micrococcus species                            18

3.5 Identification of isolates                                                                                                 19

3.5.1 Gram staining                                                                                                               19

3.5.2 Spore staining                                                                                                               19

3.6 Biochemical tests                                                                                                             19

3.7 Screening of isolate for enzyme production                                                                    21                  

3.8 Determination of Amylase activity                                                                                  22

3.8.1 Determination of temperature effect on enzyme production                                        23

3.8.2 Determination of pH effect on enzyme production                                                      23

CHAPTER FOUR

4.0 RESULTS                    ﾿ 24

CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATION ﾿             32

5.1 Discussion ﾿             32

5.2 Conclusion ﾿ 33

5.3 Recommendation ﾿ 34

REFERENCES                                                                                                                     35

APPENDIX                                                                                                                           39