Íslenskar Hverabakteríur

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Íslenskar Hverabakteríur Viðskipta- og raunvísindasvið 2014 LOK1126 og 1226 Íslenskar loftháðar hverabakteríur Sigrún Guðmundsdóttir Lokaverkefni á líftæknibraut Háskólinn á Akureyri Viðskipta- og raunvísindasvið Auðlindadeild Námskeið LOK1126 og LOK1226 Heiti verkefnis Íslenskar loftháðar hverabakteríur Verktími Nóvember 2013 – apríl 2014 Nemandi Sigrún Guðmundsdóttir Leiðbeinandi Dr. Jóhann Örlygsson Upplag 4 Blaðsíðufjöldi 78 Viðaukar Engir Fylgigögn Engin Útgáfu- og notkunarréttur Opið verkefni. Verkefnið má ekki fjölfalda, hvorki að hluta né í heild, nema með skriflegu leyfi höfundar. i Yfirlýsingar Ég lýsi því yfir að ég ein er höfundur þessa verkefnis og að það er afrakstur eigin rannsókna. _________________________ Sigrún Guðmundsdóttir Það staðfestist að verkefni þetta fullnægir að mínum dómi kröfum til prófs í námskeiðinu LOK 1126 og 1226. _________________________ Jóhann Örlygsson ii Abstract This project contains review of literature, concerned with outlined habitats, species and metabolic processes of thermophilic, aerobic bacteria and chemolithotropic that have biotechnological implications, focusing on bacteria that have been isolated in Iceland. The project summarizes the results of studies that have been associated with this task and utilization possibilities of the organism in question and their products. Iceland has favorable geothermal areas for research in this field and one conclusion of the project is that there is a great potential for further research on the diverse aerobic bacteria present in Icelandic hot springs. The project also includes a summary of the bacteria that have been detected in Iceland related to the subject and the possibilities of biotechnological utilization. Keywords: Thermophilic, aerobic, chemolithotrophic, bacteria, enzyme, biological activity iii Þakkarorð Ég þakka leiðbeinanda mínum Jóhanni Örlygssyni fyrir leiðsögnina, þarfar ábendingar, handleiðslu og ómælda þolinmæði. Einnig fyrir að hafa gefið mér tækifæri til að takast á við þetta verkefni. Samstarfsfólk mitt hjá Heilbrigðiseftirliti Suðurlands fær sérstakar þakkir fyrir stuðninginn síðustu fjögur árin og fyrir að gefa mér færi á að láta þennan draum rætast. Stelpurnar fá þakkir fyrir alla hjálpina og ekki síður félagsskapinn í gegnum þykkt og þunnt. Ekki síst fær fjölskyldan mín kærar þakkir og sérstaklega eiginmaður minn Sigurjón Valdimar Jónsson fyrir að vera ávallt til staðar þegar á þurfti að halda. Selfossi 28. apríl 2014 _________________________________ Sigrún Guðmundsdóttir kt. 041060-2639 iv Útdráttur Þetta verkefni er heimildavinna og ritgerð þar sem gerð er grein fyrir búsvæðum, uppruna og efnaskiptaferlum hitakærra, loftháðra og efnatillífandi baktería sem hafa líftæknilega skírskotun, með áherslu á bakteríur sem hafa verið einangraðar á Íslandi. Í verkefninu eru teknar saman niðurstöður úr rannsóknum sem gerðar hafa verið tengdar þessu viðfangsefni og nýtingarmöguleikum þeirra lífvera sem um ræðir og afurða þeirra. Á Íslandi eru ákjósanlegar aðstæður til rannsókna á þessu sviði og er niðurstaða verkefnisins sú að miklir möguleikar felist í enn frekari rannsóknum á þeim fjölbreyttu loftháðu hverabakteríum sem er hér að finna. Ritgerðin felur einnig í sér yfirlit yfir þær bakteríur sem hafa greinst á Íslandi og falla undir viðfangsefnið ásamt nýtingarmöguleikum. Lykilorð: Hitakærar, loftháðar, efnatillífandi, bakteríur, ensím, lífvirkni v Efnisyfirlit Yfirlýsingar ................................................................................................................................ ii Abstract ..................................................................................................................................... iii Þakkarorð .................................................................................................................................. iv Útdráttur ..................................................................................................................................... v Myndaskrá ................................................................................................................................. ix Töfluskrá ................................................................................................................................... ix 1 Inngangur ........................................................................................................................... 1 2 Efni og aðferðir .................................................................................................................. 2 3 Jarðhitasvæði ...................................................................................................................... 3 3.1 Háhitasvæði ................................................................................................................. 4 3.2 Lághitasvæði ................................................................................................................ 4 4 Hitakærar bakteríur ............................................................................................................ 6 4.1 Uppruni hitakærra baktería .......................................................................................... 8 4.2 Lífsskilyrði hitakærra baktería ................................................................................... 11 4.2.1 Loftháðar hitakærar bakteríur ............................................................................. 12 4.2.2 Hitaþol próteina og frumuhimna ........................................................................ 12 5 Efnaskipti ......................................................................................................................... 13 5.1 Glýkólýsa, sítrónusýruhringur, öndunarkeðja ........................................................... 14 5.2 Efnatillífandi, frumframleiðendur ............................................................................. 17 5.3 Næringarkröfur .......................................................................................................... 20 5.3.1 Vetni ................................................................................................................... 20 5.3.2 Brennisteinn ....................................................................................................... 21 5.3.3 Köfnunarefni ...................................................................................................... 23 vi 5.3.4 Járn og aðrir málmar .......................................................................................... 23 5.4 Ræktun baktería og seyting efna ............................................................................... 24 6 Hitakærar loftháðar bakteríur sem hafa verið einangraðar á Íslandi ................................ 25 7 Líftæknileg not hitakærra baktería ................................................................................... 28 7.1 Ensím ......................................................................................................................... 28 7.1.1 Bakteríudrepandi ensím ...................................................................................... 28 7.1.2 Niðurbrotsensím fyrir hráefni til gerjunar .......................................................... 29 7.1.3 Ensím til erfðarannsókna .................................................................................... 29 7.1.4 Önnur ensím ....................................................................................................... 30 7.2 Osmósu aðlögun ........................................................................................................ 30 7.3 Erfðafræðileg verkfæri (e. Genetic tools) .................................................................. 31 7.3.1 Lífhreinsun ......................................................................................................... 31 7.4 Rannsóknir ................................................................................................................. 33 8 Eftirfarandi hitakærar, súrefnisháðar bakteríur hafa verið einangraðar á Íslandi............. 35 8.1 Thermaceae ................................................................................................................ 35 8.1.1 Thermus ættkvíslin .............................................................................................. 35 8.1.2 Thermus islandicus ............................................................................................. 36 8.1.3 Thermus igniterrae og Thermus antranikianii ................................................... 37 8.1.4 Thermus oshimai ................................................................................................ 37 8.1.4.1 Thermus scotoductus ................................................................................... 38 8.1.5 Líftæknileg not af Thermus ................................................................................ 38 8.2 Rhodothermaceae ...................................................................................................... 39 8.2.1 Rhodothermus ættkvíslin .................................................................................... 39 8.2.2 Rhodothermus marinus ...................................................................................... 40 8.2.3 Líftæknileg not Rhodothermus ........................................................................... 40 8.3 Aquificaceae .............................................................................................................. 41 vii 8.3.1 Aquifex, Hydrogenobacter og
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