CITRIC ACID PRODUCTION FROM ORANGE PEELS USING ASPERGILLUS NIGER

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ABSTRACT


Citric acid is a ubiquitous intermediate product of metabolism and is found in practically all plants and animals. The aim of the study was to utilize orange peel as biomass for the production of citric acid using A. niger. Samples of   orange (Citrus sinensis) peels were obtained from a national citrus processing plant and was processed. The study revealed the presence of Aspergillus niger isolated from soil was able to produce citric acid from dried orange peels. The effect of inoculum concentration on citric acid production from the present study revealed that at 96hrs of incubation the inoculum concentration increased to 0.93g/L of 0.30g/L optical density while yield of citric acid from varying amounts of the fruit peel obtained were 0.42g/L, 0.66g/L, 0.73g/L, 0.81g/L and 1.04g/L respectively. In conclusion the production of citric acid using peels by Aspergillus niger has been reported to be easy, economical and eco-friendly. This method produced fairly high yield of citric acid and also it is an effective waste management method for the production of citric acid in the food and pharmaceutical industries.






TABLE OF CONTENTS


Title page                                                                                                             i

Certification                                                                                                                       ii

Dedication                                                                                                                          iii

Acknowledgement                                                                                                                                             iv

Table of contents                                                                                                             v

List of Tables                                                                                                                           viii

Abstract                                                                                                                                                                                                                                                ix


CHAPTER ONE

 

1.0 Introduction                                                                                                                      1

1.1 Aim and Objectives                                                                                                          3

1.2 Objectives                                                                                                                         3


CHAPTER TWO

2.0 LITERATURE REVIEW                                                                                              4

2.1 Citric Acid Production by Aspergillus niger (A. niger)                                                   4

2.1.1 Formation of Citric Acid                                                                                               4

2.2 Microbial Production of Citric Acid                                                                                6

2.4 Citric Acid Production Techniques                                                                                  8

2.4.1 Submerged Fermentation                                                                                              10

2.4.2 Surface Fermentation                                                                                                    11

2.4.3 Solid State Fermentation (SSF)                                                                                     11

2.5 Factors Affecting Citric Acid Production                                                                        12

2.5.1 Medium and Its Components                                                                                         12

2.6 Process parameters                                                                                                           16


CHAPTER THREE

3.0 MATERIALS AND METHODS                                                                                   19

3.1 Sample Collection                                                                                                            19

3.2 Media to be Used                                                                                                              19

3.3 Microorganisms Used                                                                                                       19

3.4 Citric Acid Production Principle                                                                                      19

3.4.1 Citric Acid Production Medium                                                                                    20

3.5 Culturing Aspergilus niger                                                                                               20

3.5.1 Estimation Citric Acid by Titrimetric Method                                                              20


CHAPTER FOUR

4.0      Results                                                                                                               21


CHAPTER FIVE

5.0   Discussion, Conclusion and Recommendation                                                                                                       25

5.1   Discussion                                                                                                                   25

5.2   Conclusion                                                                                                                  27

5.3   Recommendation                                                                                                                                                                                                                 27

References                                                                                                                  28

Appendix

 

 

 

 

 

LIST OF TABLES


  Table                                   Title                                      Page

 1                     Macroscopic and Microscopic Features of Fungal Isolate                                     22

2                      Effect of Inoculum Concentration on Citric acid Production (g/L)                     23

3                      Yield of Citric acid from varying amounts of the fruit peel                                     24

 

 


 


 

 

CHAPTER ONE


1.0     INTRODUCTION

Citric acid is a ubiquitous intermediate product of metabolism and is found in practically all plants and animals (Papagianni, 2019). The widespread presence of citric acid in animal and plant kingdom is proof of its nontoxic nature and it has high water solubility (Padvi and Pawar, 2011; Ghosh, 2013), biodegradability, palatability and is a product adjudged to be Generally Recognized As Safe (GRAS) (Nwoba et al, 2012; Bezalwar et al, 2013). It is a biotechnological and biochemical product which is most used and produced through fermentation in tones with an annual production of 1.6 million tonnes (Nadeem et al, 2010; Nwoba et al, 2012). About 70% of total citric acid produced is consumed by food industry, 12% by pharmaceutical industries and the remaining 18% consumed by other industries (Da Silva et al, 2012).

  Its applications citric acid include acidulation, preservation, anti-oxidation, flavour enhancement, plasticizer and synergistic agent (Nadeem et al, 2010; Femi-Ola and Atere, 2013; Bezalwar et al, 2013; Ghosh, 2013). Citric acid can be produced by many microorganisms and related yeast species (Pawar and Pawar, 2014). At the present day most citric acid is produced using fungi A. niger (Ali et al, 2016). The reasons for choosing A. niger over other potential citric acid producing microorganisms are; its high citric acid productivity at low pH without secretion of toxic metabolites (Nwoba et al, 2012; Haider, 2014), ease of handling (Nadeem et al, 2010), and ability to ferment a variety of cheap raw materials such as brewers spent grain (Femi-Ola and Atere, 2013), orange peel (Torrado et al, 2011), cotton waste, cane molasses, bagasse, wheat bran, coffee husk and pumpkin (Majumder et al, 2010; Kareem and Rahman., 2011; Pawar and Pawar., 2014).

Citric acid is an important commercial product, its global production has reached to 1.7 million

tons per year and its annual increasing growth rate is 5% (Faisalabad and Faisalabad, 2013). Considerable amounts of citric acid are required in several industrial processes. Approximately

70% of citric acid produced is used in the food and beverage industry as pH adjuster, flavor

enhancer, a preservative in processed food and as an acidulant in drinks (Dhandayuthapani et al,

2013). Pharmaceutical industry consumes 12% of citric acid to enhance flavors of medicines, as

an acidulant and anti-coagulator, Cosmetic industry uses citric acid in the composition of different cosmetic products (Dhillon et al, 2010). Citric acid has also found other application

such as metal cleaning, electroplating, fabric dyeing; detergent etc. (Faisalabad and Faisalabad,

2013). Citric acid production can be carried out by different groups of microorganisms. Although yeast, bacteria and other Aspergillus species can produce citric acid, Aspergillus niger (A. niger)

remained the organism of choice for commercial production, due to ease of handling, its ability to ferment a variety of cheap raw materials and produces more citric acid per time unit (Kishore

et al, 2013).

A cost reduction in citric acid production can be achieved by using cheap agricultural wastes

(Rahman, 2013). It is possible to produce citric acid by bacteria, however, not able to produce

commercially acceptable yields due to the fact that citric acid is a metabolite of energy metabolism and its accumulation rises in appreciable amounts only under conditions of drastic

imbalances (Yalcin et al, 2010). The problem in the production of citric acid by yeast is the simultaneous formation of isocitrate (Show et al, 2015). Recently, a wide range of citric acid production has been reported in response to different levels of nutrient supplementation under different articles. A. niger is capable of producing very high levels of citric acid, about 90% of the theoretical yield from a carbohydrate source. A high rate of acidogenesis in A. niger is observed only under conditions of high glycolytic metabolism and can be induced by the addition of an excess amount of sucrose or other carbohydrates (Ali, 2016). Citric acid accumulation is known to be stimulated by high sugar concentration suggesting it may be influenced by osmotic stress (Ali and Haq, 2014).


1.1 AIM AND OBJECTIVES

The aim of this study is to utilize orange peel as biomass for the production of citric acid using A. niger.


1.2 OBJECTIVES

1. To determine the effect of inoculum concentration on citric acid production

2 To yield of citric acid from varying amounts of orange peels.

3. To isolate and characterize the features of the fungal isolate

 

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