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研究生:雲妮兒
研究生(外文):Yuniar Arum Hartanti
論文名稱:篩選並鑑定海洋酵母菌以分解 Naphthalene 與燃料油
論文名稱(外文):Screening and Identification of Marine Yeast for Naphthalene and Fuel Oil Degradation
指導教授:李晏忠
指導教授(外文):Yen-Chung, Lee Ph.D
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:生物農業科技學系研究所
學門:農業科學學門
學類:農業技術學類
論文種類:學術論文
畢業學年度:103
論文頁數:53
中文關鍵詞:NaphthaleneFuel OilBiodegradationMarine YeastBiosurfactants
外文關鍵詞:NaphthaleneFuel OilBiodegradationMarine YeastBiosurfactants
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Fuel oil serves as main fuel for most of ships and industrial motor power all over the world. However, the fuel oil spill that contaminated the ecosystems is the most ominous problem today. The cleanup of spilled oil using the chemical combined with mechanical methods was never complete; therefore cause long-term ravage of ecosystem. Environmental microorganisms play the key role as final cleaner of spilled oil. Fuel oil contain large amount of polycyclic aromatic hydrocarbons (PAHs) which were quite resistant to biodegradation. So far, many bacteria species isolated from soil and seawater have been identified which capable to degrade PAHs. Yeast is one of the main populations of marine microbes, but only few strains have been demonstrated to degrade PAHs. The present study aimed to isolate and identify the marine yeast that can degrade naphthalene which occupied the most abundance component in PAHs. In preliminarily, twelve yeast strains (MY1~12) were isolated from seawater and the naphthalene degradation was tested. There are four isolates, named MY1, 8, 10, 12, showed naphthalene degradation ratio by 90%, 79%, 82%, and 78%, respectively, within 10 days reaction. Analysis of emulsification by isolates revealed that MY8 could efficiently emulsify fuel oil. In 2% MY8 cell inoculation, 0.5% of fuel oil was completely emulsified within 4 days. The result suggests that the isolated Candida sp. could produce and secrete considerable amount of biosurfactants. ITS (Internal Transcribed Spacer) sequencing and physiological analysis, MY1 was closed to Rhodosporidium fluviale which recently characterized as cesium-137 (137Cs) bio-sorbent. MY8 was identified as Candida sp. which consisted of more than 314 species of the genus. The accurate species name of the isolated Candida sp. and the kind of biosurfactant will be characterized in the future.
Fuel oil serves as main fuel for most of ships and industrial motor power all over the world. However, the fuel oil spill that contaminated the ecosystems is the most ominous problem today. The cleanup of spilled oil using the chemical combined with mechanical methods was never complete; therefore cause long-term ravage of ecosystem. Environmental microorganisms play the key role as final cleaner of spilled oil. Fuel oil contain large amount of polycyclic aromatic hydrocarbons (PAHs) which were quite resistant to biodegradation. So far, many bacteria species isolated from soil and seawater have been identified which capable to degrade PAHs. Yeast is one of the main populations of marine microbes, but only few strains have been demonstrated to degrade PAHs. The present study aimed to isolate and identify the marine yeast that can degrade naphthalene which occupied the most abundance component in PAHs. In preliminarily, twelve yeast strains (MY1~12) were isolated from seawater and the naphthalene degradation was tested. There are four isolates, named MY1, 8, 10, 12, showed naphthalene degradation ratio by 90%, 79%, 82%, and 78%, respectively, within 10 days reaction. Analysis of emulsification by isolates revealed that MY8 could efficiently emulsify fuel oil. In 2% MY8 cell inoculation, 0.5% of fuel oil was completely emulsified within 4 days. The result suggests that the isolated Candida sp. could produce and secrete considerable amount of biosurfactants. ITS (Internal Transcribed Spacer) sequencing and physiological analysis, MY1 was closed to Rhodosporidium fluviale which recently characterized as cesium-137 (137Cs) bio-sorbent. MY8 was identified as Candida sp. which consisted of more than 314 species of the genus. The accurate species name of the isolated Candida sp. and the kind of biosurfactant will be characterized in the future.
ABSTRACT ii
ACKNOWLEGDEMENT iii
ABBREVIATION iv
LIST OF CONTENTS vi
LIST OF FIGURES viii
LIST OF TABLES ix
INTRODUCTION 10
1.1 Fuel oil pollution 10
1.2 Polycyclic Aromatic hydrocarbons (PAHs) 10
1.3 Naphthalene 11
1.4 Metabolism of naphthalene in mammals 12
1.5 Biodegradation of naphthalene 12
1.6 Study purpose 13
MATERIALS AND METHODS 14
2.1 Chemicals 14
2.2 Medium preparation 14
2.3 Isolation of marine yeast 14
2.4 Growth and storage of marine yeasts culture 14
2.5 Screening of naphthalene by marine yeasts 15
2.5.1 Naphthalene degradation by marine yeast (Preliminary treatment) 15
2.5.2 Naphthalene degradation by selected marine yeast 15
2.5.3 Analysis of naphthalene by HPLC 15
2.6 Emulsification of fuel oil by marine yeast 16
2.7 Identification of the selected marine yeast 16
2.7.1 Molecular identification of selected marine yeast 16
2.7.2 Polymerase Chain Reaction (PCR) 16
2.7.3 Phylogenetic analysis 17
2.7.4 Biochemical and morphological test 17
RESULT 18
3.1 Marine fungi cultivation 18
3.2 Isolation of naphthalene degrading marine yeast 18
3.3 Determination of naphthalene degradation by marine yeast 18
3.4 Emulsification of fuel oil by marine yeast 19
3.5 Identification of marine yeast 20
DISCUSSION 21
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