Isolation Of Lipase Producing Bacteria From Palm Plantation Soil; A Case Of Optimization

ABSTRACT

Isolation and optimization of Lipase producing bacteria from palm plantation soils of Michael Okpara University of Agriculture, Umudike was carried out using culture techniques. Microorganisms of four genera were isolated from the three samples collected from the areas under study. Bacillus, Staphylococcus and Pseudomonas occurred (100%) in all the samples which suggests them to be most prevalent while Enterobacteria sp occurred in only two samples (67%). Optimization was carried out using Pseudomonas  which showed highest clearance zone in the isolation medium. Maximal lipase production was achieved at the optimum incubation time of 48hours (25.90µ/mol), temperature of 40oC (24.41 µ/mol) and pH of 7.0 (24.59 µ/mol) using olive oil as the standard substrate. The effect of various substrate for optimum production of lipase was also investigated and palm oil was observed to have the peak enzyme activity. (22.07 µ/mol) That is to say that the search for novel gene for enzyme production should be channeled to soil sample exploitation.

TABLE OF CONTENT

Preliminary Pages

Title Page ﾿ i

Certification ﾿ ii

Dedication ﾿ iii

Acknowledgement ﾿ iv

Table of Content ﾿ v

List of Tables ﾿ vi

List of Figures ﾿ viii

Abstract ﾿ ix

Chapter One

1.1 ﾿ Introduction ﾿ 1

1.2 ﾿ Aim of Study ﾿ 4

1.3 ﾿ Objective ﾿ 4

Chapter Two

2.0 ﾿ Literature Review ﾿ 5

2.1 ﾿ Lipases ﾿ 5

2.2 ﾿ Sources of Lipases ﾿ 6

Chapter Three

3.0 ﾿ Materials And Methods ﾿ 10

3.1 ﾿ Area of Study ﾿ 10

3.2 ﾿ Sample and Sampling ﾿ 10

3.3 ﾿ Isolation of Lypolytic Bacteria ﾿ 10

3.4.0 ﾿ Characterization of Lypolytic Bacteria Isolates ﾿ 11

3.4.1 ﾿ Colony Morphology and Characteristics ﾿ 11

3.4.2 ﾿ Microscopic Morphological Characteristics ﾿ 11

3.4.3 ﾿ Biochemical Reaction Test ﾿ 12

3.4.4 ﾿ Identification of Lypolytic Bacteria Isolate ﾿ 12

3.5.0 ﾿ Optimization of Parameters for Lypolytic Bacteria Activity ﾿ 12

3.5.1 ﾿ Optimization of Temperature for Lipase Activity ﾿ 12

3.5.2 ﾿ Optimization of pH for Lipase Activity ﾿ 13

3.5.3 ﾿ Optimization of Incubation Time ﾿ 13

3.5.4 ﾿ Optimization of Substrates for Lipase Activity ﾿ 13

3.6 ﾿ Extraction of Crude Enzyme ﾿ 14

3.7 ﾿ Determination of Lipase Activity ﾿ 14

Chapter Four

4.0 ﾿ Results ﾿ 15

Chapter Five

5.0 ﾿ Discussion ﾿ 26

5.1 ﾿ Conclusion ﾿ 29

5.2 ﾿ Recommendation ﾿ 30

﾿ References ﾿