STUDY OF ANTIOXIDANT- PRODUCING MICROORGANISMS IN FRUITS

ABSTRACT

Study of antioxidants producing microorganisms in fruits. Antioxidants producing microorganisms has compelling evidence that a diet rich in fruit and vegetables can help maintain a healthy lifestyle. Lactic acid bacteria was isolated from fruits (Apple, Banana and Grape). The samples were thoroughly washed, air dried and homogenized and fermented between 24-72 hours. The isolates showed clear zone on modified  de man, Rogosa and  Sharpe (Mrs). Catalase negative and Gram positive bacteria such as  Lactococcus, Pediococcus, Streptococcus, Enterococcus were used. The proteolytic activity of these organisms were 6.67±0.58 (Lactococcus), 8.38 ±1.15 for  (Pediococcus), 10.33±0.58 (Streptococcus). The chelating activities was 17.78±0.29 for  Lactococcus, 12.62 ±0.17 (Pediococcus), 10.02±0.25 (Enterococcus) 17.22± 0.26 (Streptococcus). The scavenging activity was 27.73 ±0.28 for Lactococcus, 31.39 ±0.35 for Pediococcus, 18.81±0.18 for Enterococcus and 21.10±0.41 for (Streptococcus). This study emphasis that the consumption of fruits will be healthful practices as it contains natural antioxidants which will remove free radicals.

TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgement iv

Table of Contents v

List of Tables vii

List of Figures

Abstract             viii

CHAPTER ONE -    INTRODUCTION 1

1.1 Introduction 1

1.2 Dietary Antioxidants 2

1.3 Natural Antioxidants and its Benefits 2

CHAPTER TWO   -   LITERATURE  REVIEW 4

2.1 Historical Review 4

2.2 Health Benefits of Natural Antioxidants 4

2.3 Natural antioxidants and its Benefits 5

2.4 Antioxidants and its Effects 6

2.5 Importance of Antioxidant in the Body 7

2.6 Microbiological Overview of the Dairy Products 7

2.7 Lactic Acid Bacteria (LAB) 8

2.8 Antioxidant Foods 9

2.9 LAB in Mycotoxin Detoxification 9

2.10 Animal Products Fermentation-NONO 10

2.11 Other Sources of Fermented Foods with Natural Antioxidant           11

CHAPTER THREE - MATERIALS AND METHODS 12

3.1 Materials 12

3.2 Isolation of Bacteria 12

3.3 Characterization  and  Identification of  LAB 12

3.3.1 Biochemical test 13

3.3.2 Catalase test 13

3.4 Identification of LAB Isolates 13

3.5 Microscopic Characteristics        13-14

3.6 Protein Hydrolysis 14

3.6.1 Preparation of Skim Milk Agar and Cultures 14

3.6.2 Measurements of Protein Hydrolysis 14

3.6.3 Selection of Lactic Acid Bacteria Isolates        14-15

3.7 Preparation of Whey Fraction from Fermented Milk 15

3.8 Determination of Antioxidant Activity 15

3.9 Scavenging of 1,1-diphenyl-2-picrylyhydrazyl (DPPH) Free  Radical 15

3.10 Ferrous Chelating Activity (FCA) 15

CHAPTER FOUR -  RESULTS        17-23

CHAPTER FIVE – DISCUSSION, CONCLUSION AND RECOMMENDATION 24

5.1 Conclusion 25

5.2 Recommendation        25-26

References

Appendixes

LIST OF TABLES

Table 1: Morphological and Biochemical  characteristics  of bacterial isolates. 18

LIST OF FIGURES

Figure Title Page

1:  Shows the occurance of LAB in fruits for antioxidant production 19

2: Shows the protein hydrolysis 20

3: Shows the scavanging activity 21

4: Shows the Chelating Activities of LAB  isolates 22

5: Showing the Standard Deviation of Lab Isolates   23

CHAPTER ONE

INTRODUCTION

1.3 Introduction

There is compelling evidence that a diet rich in fruit and vegetables can help maintain a healthy lifestyle. Current recommendations are that everyone should eat at least five portions of a variety of fruit and vegetable each day to reduce the risk of chronic illness such as cancer and Cardiovascular diseases-. Endogenous antioxidant mechanisms, including enzymatic defences, contribute to their efficient removal. However, when such defence mechanisms fail or are insufficient to lope, these free radicals trigger chain reactions resulting in the rapid oxidation of target molecules. The increases exposure to free radicals, known as oxidative/nitrosative stress, can lead to DNA, lipid and protein damage and potentially increases the risk of cardiovascular illness and cancer (Evans et al, 2013). Antioxidants producing foods namely fruits has receives increasing attention as potential ingredients for health.

- Promoting   functional foods targeted at direct.

-  Related chronic diseases, such as cardiovascular disease, diabetes type II and others. These peptides are inactive within the sequence of the parent protein molecule and it can be released through:- enzymatic hydrolysis by digestive enzymes, through fermentation of milk with proteolytic starter cultures and proteolyses  by enzymes  derived from microorganisms or plant  (Hannu, 2009). The fermentation of milk by lactic acid bacterial (LAB) releases  a large number  of peptides and amino acids with different biological  actions, such  as Angiotensin converting enzyme (ACE) inhibitory (Nielsen  et al, 2009) immune  modulatory   (Coste et al,1992) and antioxidant  activities  (Pena-  Rasmos et al, 2004).

Pairote et al., (2007) have reported that almost organism   including human and animals are well protected   against   free radical damage by endogenous enzymes such as superoxide dismutase and  catalase, or compounds such as   ascorbic acid, tocophenol and glutathione but when this mechanism become imbalance by many factors such as aging or detorioration of physiological functions, pathological stress may occur. Therefore, it is imperative that the supplementation of antioxidants or enhancement   of antioxidant in food   for human and animals may be used to reduce oxidative sties in body caused by free radicals (Pairote  et al.,  2007).

1.4 Dietary Antioxidants

Many components in foodstuffs can contribute to their health promoting effects, including micronutrinets such as Vitamins C and E and Minerals like Selenium, Carotenoids, glucosinolates, Indoles, Isothiocyanates, fiber, folic acid etc. The antioxidant capacity of several natural compounds has been thought to be a key factor in the negative correlation between a fruit and vegetable-rich diet and chronic illness, potentially through their ability to supplement cellular defence systems (Emillie, 2008). Major sources of antioxidant in the human diet are varied and include cereals, fruits vegetables, chocolate, oils and beverages such as tea, coffee, wine and fruit juices. The antioxidant profile of plants differ according to the species and is also influenced by farming (organic vs traditional), packaging, transport, storage as well as cooking and commercial processing (Emillie, 2008).

1.3 Aims and Objectives

The purpose of this project therefore is;

- To select fruits producing antioxidants.

- To investigate the relationship of some bacteria with their antioxidant producing fruits.

- To determine protein hydrolysis by their bacteria.

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