Pigment analysis in Sphingomonas glacialis strain S2U11 and its light exposure effect
Tekijät
Päivämäärä
2023Tekijänoikeudet
© The Author(s)
Hapellisissa oloissa happea tuottamattomat fotosynteettiset (AAP-) bakteerit saavat kemiallista energiaa hyödyntämällä valoa happea tuottamattomassa fotosynteesissä. Hapellinen ympäristö on tarpeellinen bakteeriklorofyllien ja karotenoidien valmistukseen, jotka absorboivat valoa valonkeräyskomplekseissa, mistä valon virittämä energia siirretään reaktiokeskuksiin eksitaatioenergiansiirto-prosessilla. Bakteeriklorofylli absorboi valoa infrapuna-alueelta ja karotenoidit 400–550 nm alueelta. Karotenoidit voivat siirtää viritysenergian bakteeriklorofylleille. Toisaalta, karotenoidit suojelevat komplekseja reaktiiviselta hapelta poistamalla ylimääräisen viritysenergian. AAP-bakteereja sekä niiden pigmenttien tutkimus on vähäistä, tämä tutkimus kehittää menetelmää pigmenttien analysointiin. Pigmentit eluoitiin Sphingomonas glacialis -bakteereista. Identifiointi tehtiin korkean erottelykyvyn nestekromatografian retentioajoilla sekä absorptiospektreillä. Fotosynteettisten pigmenttien kartoitus tehtiin fluoresenssieksitaatiolla. Valo-olosuhteiden muutokset vaikuttivat pigmentteihin ja runsain kasvu tapahtui pimeässä. Pigmenttejä löytyi runsaasti, mutta yksikään ei ollut fotosynteettinen. Pigmenttien analysointimenetelmä on toimiva ja käytettävissä muihin AAP-bakteereihin, mutta karotenoidien tunnistamisessa on vielä käytännön haasteita.
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Aerobic anoxygenic photosynthetic (AAP) bacteria gain chemical energy from light through non-oxygen producing photosynthesis. The presence of the oxygen is required to produce pigments, bacteriochlorophyll a (BChla a) and carotenoids. During photosynthesis pigment constituting light harvesting (LH) complexes gather absorbed light and transfer it to reaction centre (RC) as excitation energy with process called singlet state energy transfer. BChl a is an essential pigment to both the LH complex and RC and its main light absorption area is in near infrared area. Photosynthetic carotenoids absorb light from 400 to 550 nm area and transfer it to the BChls. Carotenoids can act as photoprotectors and quench the excess light energy before it creates high energetic singlet oxygen, that is detrimental to BChl synthesis. The research of AAP bacteria, especially their pigment content, is scarce. This thesis works to find possible method for the pigment analysis and identification. The pigments were extracted from Sphingomonas glacialis and identified with high performance liquid chromatography retention times and absorption spectra. The presence of photosynthetic carotenoids was studied with fluorescence excitation. The light exposure examination addressed effect of light changes to the pigment composition. Pigment synthesis takes place in the darkness. Photosynthetic carotenoids were not present in Sphingomonas glacialis, but multiple carotenoids were found. Identification of the pigments is challenging but the method for pigment analysis is functional and can be utilized in other carotenoid containing AAP bacteria.
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Asiasanat
BChl a HPLC carotenoids fluorescence excitation singlet state energy transfer anoxygenic photosynthesis anoxygenic photosynthetic bacteria yhteyttäminen karotenoidit bakteerit valotus pigmentit (värijauheet) fluoresenssi imeytyminen photosynthesis bacteria exposure to light pigments fluorescence absorption
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