PRODUCTION OF PROTEASE BY ASPERGILLUS FLAVUS IN SOLID STATE FERMENTATION

PRODUCTION OF PROTEASE BY ASPERGILLUS FLAVUS IN SOLID STATE FERMENTATION

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 that Aspergillus flavus was able to produce extracellular protease enzymes important in the decomposition of protein materials.

                                      CHAPTER ONE

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:

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

 

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How To Write Chapter Three Of Your Research Project (Research Methodology)

  • Methodology In Research Paper


    Chapter three of the research project or the research methodology is another significant part of the research project writing. In developing the chapter three of the research project, you state the purpose of research, research method you wish to adopt, the instruments to be used, where you will collect your data, types of data collection, and how you collected it.

    This chapter explains the different methods to be used in the research project. Here you mention the procedures and strategies you will employ in the study such as research design, study design in research, research area (area of the study), the population of the study, etc. You also tell the reader your research design methods, why you chose a particular method, method of analysis, how you planned to analyze your data.

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    You can choose a survey design when you want to survey a particular location or behavior by administering instruments such as structured questionnaires, interviews, or experimental; if you intend manipulating some variables.

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    Method vs Methodology

    I think the most appropriate in methods versus methodology is to think in terms of their inter-connectedness and relationship between both. You should not beging thinking so much about research methods without thinking of developing a research methodology.

    Metodologia or methodology is the consideration of your research objectives and the most effective method and approach to meet those objectives. That is to say that methodology in research paper is the first step in planning a research project work.

    Design Methodology: Methodological Approach

    Example of methodology in research paper, you are attempting to identify the influence of personality on a road accident, you may wish to look at different personality types, you may also look at accident records from the FRSC, you may also wish to look at the personality of drivers that are accident victims, once you adopt this method, you are already doing a survey, and that becomes your metodologia or methodology.

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    Click here to complete this article - How To Write Chapter Three Of Your Research Project (Research Methodology)

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