Production of protease by aspergillus flavus in solid state fermentation
Table Of Contents
Chapter ONE
INTRODUCTION
- 1.1Introduction
- 1.2Background of Study
- 1.3Problem Statement
- 1.4Objective of Study
- 1.5Limitation of Study
- 1.6Scope of Study
- 1.7Significance of Study
- 1.8Structure of the Research
- 1.9Definition of Terms
Chapter TWO
LITERATURE REVIEW
- 2.1Overview of Protease Enzyme
- 2.2Sources of Protease
- 2.3Production of Protease Enzyme
- 2.4Aspergillus flavus: Characteristics and Importance
- 2.5Solid-State Fermentation Process
- 2.6Factors Affecting Protease Production
- 2.7Applications of Protease Enzyme
- 2.8Enzyme Purification Techniques
- 2.9Enzyme Stability and Storage
- 2.10Recent Developments in Protease Research
Chapter THREE
SYSTEM DESIGN AND IMPLEMENTATION
- 3.1Research Methodology Overview
- 3.2Selection of Aspergillus flavus Strain
- 3.3Preparation of Solid-State Fermentation Media
- 3.4Inoculation and Incubation Process
- 3.5Sampling and Analysis Methods
- 3.6Optimization of Culture Conditions
- 3.7Enzyme Extraction and Purification
- 3.8Enzyme Activity Assays
Chapter FOUR
SYSTEM TESTING AND EVALUATION
- 4.1Analysis of Protease Production Results
- 4.2Effects of Nutrient Composition on Enzyme Yield
- 4.3Influence of Fermentation Parameters on Protease Activity
- 4.4Comparison of Protease Production in Solid-State vs. Submerged Fermentation
- 4.5Enzyme Characterization Studies
- 4.6Factors Affecting Enzyme Stability
- 4.7Industrial Applications of Aspergillus flavus Protease
- 4.8Future Prospects and Recommendations
Chapter FIVE
SUMMARY, CONCLUSION AND RECOMMENDATIONS
- 5.1Summary of Findings
- 5.2Conclusions
- 5.3Implications of the Research
- 5.4Recommendations for Future Studies
- 5.5Closing Remarks
Project Abstract
<p> 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. <br></p>
Project Overview
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</p><p><strong>1.0 INTRODUCTION</strong></p><p><strong>1.1 Background of the study</strong></p><p>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.</p><p>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.</p><p>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.</p><p>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).</p><p><strong>1.2 Aim of the study</strong></p><p>The aim of this study was to produce protease from Aspergillus flavus using wheat bran as a substrate under Solid State Fermentation.</p><p><strong>1.3 Objectives of the study</strong></p><p>The objectives of the study include:</p><p>To isolate Aspergillus flavus from spoilt bread in Gwagwalada.</p><p>To determine the frequencies of occurrence of the isolated Aspergillus flavus from spoilt bread using simple percentages.</p><p>To determinethe proteolytic potential of the isolated fungi using basal medium.</p><p>To determine the quantity of the protease enzyme produced by the isolated fungi using spectrophotometer.</p>
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