Isolation and screening of fungi from the rhizosphere of moringa oleifera for amylase and protease production

ABSTRACT
The present study is to isolate fungi from rhizosphere of Moringa oleifera and screen them for the production of some very important enzymes such as amylase and protease. Twenty fungi were isolated from the soil sample from five different farms using pour plate method. The isolates were then identified based on their colonial morphology and microscopic features in wet mount and Lactophenol Cotton Blue staining. All the isolates were screened for amylase and protease production by starch agar plate method and casein agar plate method respectively. A total of seven isolates were identified up to the species level, while thirteen isolates were identified up to the genus level. The isolate MCS 16 having the highest amylase and protease ability was identified as Aspergillus spp 3. Aspergillus nidulans, Aspergillus niger, Aspergillus terreus Penicillium spp 1, Aspergillus fumigatus, Alternaria spp 1, Rhizopus spp 1, Rhizopus stolonifer, and Aspergillus spp 5, were found in all the five farms where the soil sample were collected with a frequency rate of 25%. The study revealed that rhizosphere of Moringa oleifera can be a very good source of fungi capable of producing enzymes

TABLE OF CONTENTS
TITLE PAGE
Certification……………………………………………………………................i
Dedication………………………………………………………………………...ii
Acknowledgements..……………………………………………………………...iii
Abstract……………………………………………………………………………iv

CHAPTER ONE
1.0 INTRODUCTION………………………………………………................1
1.1    Fungi………………………………………………………………….1
1.2   Amylases……………………………………………………………...2
          1.3   Proteases……………………………………………………………….3
 1.4   Rhizosphere……………………………………………………………4
 1.5   Moringa oleifera………………………………………………………4
 1.6   Justification……………………………………………………………4
 1.7   Aim……………………………………………………………………5 
        1.7.1 Objectives…………………………………………………….5

CHAPTER TWO
2.0 LITERATURE REVIEW…………………………………………………..6
2.1   Functions of Amylase…………………………………………………8
2.1.1   Bread and Chapatti industry…………………………………8
2.1.2   Textile industry……………………………………………...9
2.1.3   Sugar and Glucose industry………………………………….9
2.1.4  Detergent, Building product and feed industries……………10
2.1.5  Paper industry………………………………………………..10
2.1.6   Alcohol industry……………………………………………..10
2.1.7   Chocolate industry…………………………………………...11
2.1.8   Feed industry…………………………………………………11
2.2   Sources of proteases…………………………………………………12
  2.2.1    Animals………………………………………………..….....12
  2.2.2    Fungi………………………………………………………...12
  2.2.3    Plants………………………………………………………..13
  2.2.4    Bacteria……………………………………………………..13
  2.2.5    Viruses……………………………………………...............13
 2.3   Applications/uses of proteases……………………………………….13
  2.3.1    Food industry………………………………………………..13
  2.3.2    Baking industry……………………………………………..14
  2.3.3    Leather industry……………………………………………..14
  2.4 Sources of Enzymes…………………………………………………14
 2.5 Sources of Microbial Enzymes……………………………………...16
2.5.1 Advantages and disadvantages of microbial enzymes……….17
2.6 Rhizosphere and enzyme production………………………………..18
     2.6.1 Rhizospheric organisms………………………………………19
2.7 Reason for search for more Enzyme Producing strains……………..20
2.8     Moringa oleifera…………………………………………………….20

CHAPTER THREE
3.0 MATERIALS AND METHODS…………………………………………..21
 3.1 Equipment and reagents used……………………………………….21
3.2 Collection of samples……………………………………………….21
3.3 Isolation of fungi……………………………………………………21
3.4 Identification of the isolates…………………………………………22
3.5 Screening for amylase production…….……………………………22
3.6 Screening for Protease………………………………………………23

CHAPTER FOUR
4.0 RESULTS………………………………………………………………….24
 4.1 Identification of the fungi isolates………………………..…………24
 4.2 Frequency of occurrence of the fungal species in each farm……….30 
 4.3 Screening the fungal species for amylase and protease…………….30
 4.4 Amylase production ability…………………………………………30
 4.5 Protease production ability………………………………………….30

CHAPTER FIVE
 DISCUSSION…………………………………………………………….34
 CONCLUSION……………………………………………………………38
 RECOMMENDATIONS………………………………………………….39
REFERENCES………………………….....................................................40
 APPENDIX………………………………………………………………..51