Determination of Oxygen Respiration Rates in Wetted Developmentally Arrested Spores of Streptomyces Species
链霉菌属物种潮湿抑制发育孢子的氧气呼吸率测定
发布: 2014年06月20日第4卷第12期 DOI: 10.21769/BioProtoc.1153 浏览次数: 9334
评审: Anonymous reviewer(s)
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Sep 2013
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Abstract
Streptomyces species produce spores, which, while not as robust as endospores of Bacillus or Clostridium species, are capable of surviving for months or even years (Hopwood, 2006). During this time these spores remain viable, surviving by slowly degrading internal stores of carbon compounds, such as the carbohydrate trehalose. To enable metabolism to continue they must have access to an electron acceptor that allows the removal of the reducing equivalents that accumulate through metabolic activity. The most commonly used acceptor is oxygen. We describe the quantitative measurement of oxygen respiration rates by developmentally arrested spores of the streptomycete Streptomyces coelicolor (Fischer et al., 2013).
Keywords: Cytochrome oxidase (细胞色素氧化酶)
Oxygen reduction (氧还原)
Spores (孢子)
Non-invasive optical oxygen-sensing (非侵入式光学氧传感)
Streptomyces coelicolor (天蓝色链霉菌)
Materials and Reagents
- Freshly harvested Streptomyces spores in water
- Soya flour (from local supermarket)
- D-mannitol (Sigma-Aldrich, catalog number: 63560 )
- Agar-agar (Kobe I) (Roth North America)
- 3-(N-morpholino) propanesulfonic acid (MOPS) buffer (Roth North America)
- Chloramphenicol (Merck KGaA)
- Sodium hydroxide pellets (Roth North America)
- MOPS-buffer (see Recipes)
- SFM agar (see Recipes)
Equipment
- Cotton wool (standard issue from local Pharmacy) for spore preparation
- 0.22 µm pore-size filters (PVDF) (Sartorius AG)
- 30 °C rotary shaker
- Baffled Erlenmeyer flasks (500 ml) (Glasgerätebau Ochs, Laborfachhandel e. K., catalog number: 100500 )
- Standard-sized plastic Petri dishes for bacterial growth and spore preparation
- Gas-tight glass Hungate tubes (16 ml) for anaerobic work with butyl rubber septa (Glasgerätebau Ochs, Laborfachhandel e. K., catalog number: 1020471 )
- Cooled table-top centrifuge (e.g. Eppendorf)
- Needles (0.6 mm gauge) (B. Braun Melsungen AG)
- Water bath or heating block (Biometra)
- Spectrophotometer capable of measuring absorption in the visible range
- Optical ‘FirestingO2 oxygen meter’ (Pyro Science GmbH)
- Contactless optical oxygen ‘sensor spot’ (Pyro Science GmbH)
- ‘Bare fibre’ (Pyro Science GmbH)
- Adjustable mini magnetic stirrer (e.g. IKA) (10 mm x 0.3 mm magnetic stirrer bar) (e.g. Roth North America)
Software
- Firesting Logger Software (Pyro Science GmbH)
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Fischer, M., Falke, D. and Sawers, R. G. (2014). Determination of Oxygen Respiration Rates in Wetted Developmentally Arrested Spores of Streptomyces Species. Bio-protocol 4(12): e1153. DOI: 10.21769/BioProtoc.1153.
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分类
微生物学 > 微生物生物化学 > 其它化合物
微生物学 > 微生物新陈代谢 > 营养运输
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