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Production of Protease by Aspergillus Flavus in Solid State Fermentation

Production of Protease by Aspergillus Flavus in Solid State Fermentation

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DEDICATION

This research material, titled “Production of Protease by Aspergillus Flavus in Solid State Fermentation” is dedicated to God for His boundless grace and guidance. It is also a tribute to all computer enthusiasts whose contributions made my research journey smoother and enriched my documentation process, making the experience truly fulfilling.




ACKNOWLEDGEMENT

I am profoundly grateful to everyone who contributed to the successful completion of this project. I am especially grateful to my Supervisor (Name), the Head of Department (Name), and the Lecturers in the Department of Microbiology for their invaluable guidance and support. I also acknowledge the contributions of authors and scholars whose works on Production of Protease by Aspergillus Flavus in Solid State Fermentation provided essential insights. Special thanks go to my study area (and any funding organizations, if applicable) for their financial assistance. I am equally thankful to stakeholders, including mentors, teachers, and colleagues, for their encouragement and support. Finally, I deeply appreciate my family and friends for their patience and unwavering support throughout this journey. Your contributions have been instrumental in making this research a reality.




PRELIMINARY PAGES


CHAPTER ONE

INTRODUCTION


    CHAPTER TWO

    LITERATURE REVIEW

    • 2.1 Introduction

    CHAPTER THREE

    MATERIALS AND METHODS

    • 3.1 Introduction

    CHAPTER FOUR

    RESULTS AND DISCUSSION

    • 4.1 Introduction
    • 4.2 Results
    • 4.3 Discussion of Findings

    CHAPTER FIVE

    SUMMARY, CONCLUSION AND RECOMMENDATION

    • 5.1 Introduction
    • 5.2 Summary
    • 5.3 Conclusion
    • 5.4 Recommendation

    REFERENCES



    ABSTRACT

    The study for the production of protease from Aspergillus flavus using wheat bran as substrate under Solid State Fermentation was conducted in University of Abuja, Department of Microbiology. Aspergillus flavus was isolated from spoilt bread and was identified on the basis of the morphological assessment such as macroscopic and microscopy. Among the characteristics used includes: colonial characteristics such surface appearance, texture and colour of the colonies. The protease activity increased with increase in the fermentation periods.

    The quantities of the protease enzyme produced by the Aspergillus flavus in the basal medium were measured using UV-Spectrophotometer and the result is shown in Table 5. The protease activity was found to be higher at day 7 than day 5 and 3 with 4.51±10.06 proteaase Unit/mL, 8.63±0.12 U/mL and 18.93±1.20 AU/mL respectively. The extracellular protease produced by Aspergillus flavus isolated from spoilt bread in Gwagwalada were not significantly different at P= 0.05 level of significance. The study demonstrated thatAspergillus flavus wasable to produce extracellular protease enzymes important in the decomposition of protein materials.



    Production of Protease by Aspergillus Flavus in Solid State Fermentation


    1.0 Introduction

    1.1 Background of the study

    Protease constitutes a large and complex group of enzymes that plays an important nutritional and regulatory role in nature. Proteases are (physiologically) necessary for living organisms; they are ubiquitous and found in a wide diversity of sources. Protease is the most important industrial enzyme of interest accounting for about 60% of the total enzyme market in the world and account for approximately 40% of the total worldwide enzyme sale (Godfrey and West, 1996; Chouyyok et al., 2005). They are generally used in detergents (Barindra et al., 2006), food industries, leather, meat processing, cheese making, silver recovery from photographic film, production of digestive and certain medical treatments of inflammation and virulent wounds (Rao et al., 1998; Paranthaman et al., 2009). They also have medical and pharmaceutical applications.

    Microbial proteases are degradative enzymes which catalyze the total hydrolysis of proteins (Raju et al., 1994; Haq et al., 2006). The molecular weight of proteases ranges from 18 − 90 kDa (Sidney and Lester, 1972). These enzymes are found in a wide diversity of sources such as plants, animals and microorganisms but they are mainly produced by bacteria and fungi. Microbial proteases are predominantly extracellular and can be secreted in the fermentation medium.

    In the production of protease, it has been shown to be inducible and was affected by the nature of the substrate used in fermentation. Therefore, the choice of an appropriate inducing substrate is of great importance. Different carbon sources such as wheat bran, rice straw, rice bran, cotton and bagasse have been studied for the induction and biosynthesis of protease. However, wheat bran is a superior carbon source for the production of protease by Aspergillus flavus. So the further studies were carried out by using wheat bran as carbon source.

    The use of agro-industrial residues as the basis for cultivation media is a matter of great interest, aiming to decrease the costs of enzyme production and meeting the increase in awareness on energy conservation and recycling (Singh et al., 2009). Major impediments to the exploitation of commercial enzymes are their yield, stability, specificity and the cost of production. New enzymes for use in commercial applications with desirable biochemical and physiochemical characteristics and low production cost have been focus of much research (Kabli, 2007). Solid state fermentation (SSF) was chosen for the present research because it has been reported to be of more grated productivity than that of submerged fermentation (Ghildyal et al., 1985; Hesseltine, 1972). Economically, SSF offers many advantages including superior volumetric productivity, use of simpler machinery, use of inexpensive substrates, simpler downstream processing, and lower energy requirements when compared with submerged fermentation (Paranthaman et al., 2009).


    1.2 Aim of the study

    The aim of this study was to produce protease from Aspergillus flavus using wheat bran as a substrate under Solid State Fermentation.


    1.3 Objectives of the study

    The objectives of the study include:

    1. To isolate Aspergillus flavus from spoilt bread in Gwagwalada.
    2. To determine the frequencies of occurrence of the isolated Aspergillus flavus from spoilt bread using simple percentages.
    3. To determine the proteolytic potential of the isolated fungi using basal medium.
    4. To determine the quantity of the protease enzyme produced by the isolated fungi using spectrophotometer.

    CHAPTER TWO

    2.0 Literature Review

    2.1 Introduction

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