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Mycoremediation of Hydrocarbon-Contaminated

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Mycoremediation of Hydrocarbon-Contaminated Natural Sciences International Bachelor 1st semester project Mycoremediation of hydrocarbon-contaminated brownfield sites using Pleurotus ostreatus Group 4 Thor Ekow Sarquah-Djurhuus Torben Callesen Katrine Jakobsen Florin Krijom Signe Skou Supervisor: Lauren Seaby Abstract In urban areas, contamination of soil with petroleum hydrocarbons restricts the amount of land available for use due to the human health hazards posed by the toxic properties of the contaminant compounds. Mycoremediation is a form of bioremediation in which the degradative abilities of fungi are utilized to remove or neutralize harmful contaminants present in the soil and ground water. This project investigates the potential of the white rot fungi Pleurotus ostreatus to remediate hydrocarbon- contaminated brownfield sites, in other words, to clean up oil spills. This is primarily done through literature based research, as well as a review of relevant case studies, and an expert interview with a mycoremediation company. P. ostreatus is relevant for hydrocarbon contamination because it feeds on organic carbon in the form of wood-lignocellulose by degrading lignin using the non-specific extracellular enzymes laccase, manganese peroxidase and versatile peroxidase, which also break down polyaromatic hydrocarbons (PAHs) due to the similarity in molecular structure to lignin. Since most native microflora and simple bacterial remediation techniques are able to degrade lower molecular weight hydrocarbons, the benefit lies primarily in the ability of P. ostreatus to degrade the more recalcitrant, higher molecular weight components, such as >5-ring PAHs, making them more accessible to other decomposers as well as the fungus. This is supported by numerous in situ and in vitro studies, which show that the use of P. ostreatus in cooperation with soil microorganism results in between 40 and 90% of degradation of total hydrocarbons present. Site conditions such as physical environmental factors and the presence of co-contaminants, such as certain heavy metals, play a significant role in the efficiency of remediation. Ex situ mycoremediation is time wise more efficient than in situ methods, but involves higher costs. Compared to bioremediation with bacteria, P. ostreatus needs less attention in regards to maintenance once it has been implemented, decreasing cost and labor requirements. In conclusion, mycoremediation using P. ostreatus has high potential for contribution to the bioremediation of brownfield sites. Contents Abstract ............................................................................................................................................ 1. Introduction.................................................................................................................................. 1 1.1. Research Question: .................................................................................................................. 2 1.1.1. Sub questions: ................................................................................................................... 2 1.2. Hypotheses: ............................................................................................................................ 2 2. Method ......................................................................................................................................... 2 3. Theory .......................................................................................................................................... 3 3.1. Brownfield sites ...................................................................................................................... 3 3.1.1. Definition ......................................................................................................................... 3 3.1.2. Urban effects of brownfield sites ......................................................................................... 4 3.1.3. Occurrence of contaminants ................................................................................................ 4 3.1.4. Contaminant nature and behavior ........................................................................................ 5 3.1.5. Toxicity ............................................................................................................................ 9 3.2. History: Bioremediation ......................................................................................................... 11 3.3. History: Mycoremediation ...................................................................................................... 12 3.4. Introduction to P. ostreatus ..................................................................................................... 14 3.4.1 Name and taxonomy ......................................................................................................... 14 3.4.2. Anatomy ........................................................................................................................ 14 3.4.3. Lifecycle ........................................................................................................................ 15 3.5. P. ostreatus as a decomposer ................................................................................................... 16 3.5.1. Saprotrophic Fungi .......................................................................................................... 16 3.5.2. Degradation of hydrocarbons ............................................................................................ 17 3.6. Fungal enzymes and the chemical degradation of lignin ............................................................. 18 3.6.1. Laccases vs Peroxidases ................................................................................................... 18 3.6.2. Laccase .......................................................................................................................... 18 3.6.3. Haem-peroxidases ........................................................................................................... 20 3.6.3.1. Manganese peroxidase ................................................................................................... 21 3.6.3.1. Versatile peroxidase ...................................................................................................... 21 3.7. P. ostreatus cultivation ........................................................................................................... 21 3.7.1. Media for cultivation ........................................................................................................ 23 3.8. Growth conditions and factors ................................................................................................. 23 3.8.1. Temperature.................................................................................................................... 23 3.8.2. Carbon/Nitrogen ratio ...................................................................................................... 24 3.8.3. Heavy Metals .................................................................................................................. 26 3.8.4. Other growth requirements ............................................................................................... 27 3.9. Other bioremediation technologies ........................................................................................... 27 3.9.1. Microorganisms: Bacteria ................................................................................................. 28 3.10. Application of bioremediation ............................................................................................... 29 3.10.1 In Situ ........................................................................................................................... 29 3.10.2. Ex situ .......................................................................................................................... 30 3.10.3. Soil Analysis ................................................................................................................. 31 3.11. Application of mycoremediation ............................................................................................ 33 4. Analysis ..................................................................................................................................... 34 4.1. Suitability of Mycoremediation ............................................................................................... 34 4.1.1. Degradative capability of organisms .................................................................................. 35 4.1.2. Rate of degradation and bioavailability .............................................................................. 35 4.1.3. Harmful byproducts ......................................................................................................... 37 4.1.4. Appropriate site conditions for biodegraders (in situ) ........................................................... 39 4.1.5 Mycoremediation in controlled conditions ........................................................................... 41 4.1.6. Economic viability ........................................................................................................... 41 4.2. Email interview: Fungi Perfecti ............................................................................................... 44 4.2.1. Barriers for implementation .............................................................................................
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