Team:Bielefeld-Germany/Results/Summary
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After the measurement of activity of BPUL we made a scale-up and fermented ''E. coli'' KRX with <partinfo>BBa_K863000</partinfo> in Braun Biostat B with a total volume of 3 L. Agitation speed, pO<sub>2</sub> and OD<sub>600</sub> were determined and illustrated in the following graph. We got a long lag phase of 2 hours due to a relativly old preculture. The cell growth caused a decrease in pO<sub>2</sub> and after 3 hours the value fell below 50 %, so that the agitation speed was increased automatically. After 8,5 hours the deceleration phase started and therefore the agitation speed was descreased. The maximal OD<sub>600</sub> of 3,53 was reached after 10 hours. The cells were harvested after 11 hours. | After the measurement of activity of BPUL we made a scale-up and fermented ''E. coli'' KRX with <partinfo>BBa_K863000</partinfo> in Braun Biostat B with a total volume of 3 L. Agitation speed, pO<sub>2</sub> and OD<sub>600</sub> were determined and illustrated in the following graph. We got a long lag phase of 2 hours due to a relativly old preculture. The cell growth caused a decrease in pO<sub>2</sub> and after 3 hours the value fell below 50 %, so that the agitation speed was increased automatically. After 8,5 hours the deceleration phase started and therefore the agitation speed was descreased. The maximal OD<sub>600</sub> of 3,53 was reached after 10 hours. The cells were harvested after 11 hours. | ||
- | [[File:Bielefeld2012_BPUL3LFermentation.jpg|600px|thumb|center|Fermentation of ''E. coli'' KRX with <partinfo>BBa_K863000</partinfo> (BPUL) in Braun Biostat B, scale: 3 L, [https://2012.igem.org/Team:Bielefeld-Germany/Protocols/Materials#Autoinduction_medium autoinduction medium] + 60 µg/mL chloramphenicol, 37 °C, pH 7, agitation on cascade to hold a pO<sub>2</sub> of 50 %, OD<sub>600</sub> taken every 30 minutes.]] | + | [[File:Bielefeld2012_BPUL3LFermentation.jpg|600px|thumb|center|Figure 1: Fermentation of ''E. coli'' KRX with <partinfo>BBa_K863000</partinfo> (BPUL) in Braun Biostat B, scale: 3 L, [https://2012.igem.org/Team:Bielefeld-Germany/Protocols/Materials#Autoinduction_medium autoinduction medium] + 60 µg/mL chloramphenicol, 37 °C, pH 7, agitation on cascade to hold a pO<sub>2</sub> of 50 %, OD<sub>600</sub> taken every 30 minutes.]] |
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+ | [[File:Bielefeld2012_BPUL3LChromatogramm.jpg|600px|thumb|center|Figure:2]] | ||
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- | [[File:Bielefeld2012_BPUL6LFermentation.jpg|600px|thumb|center|Fermentation of ''E. coli'' KRX with <partinfo>BBa_K863005</partinfo> (BPUL) in Bioengineering NFL22, scale: 6 L, [https://2012.igem.org/Team:Bielefeld-Germany/Protocols/Materials#Autoinduction_medium autoinduction medium] + 60 µg/mL chloramphenicol, 37 °C, pH 7, agitation increased when pO<sub>2</sub> was below 30 %, OD<sub>600</sub> taken every hour. ]] | + | [[File:Bielefeld2012_BPUL6LFermentation.jpg|600px|thumb|center|Figure 3: Fermentation of ''E. coli'' KRX with <partinfo>BBa_K863005</partinfo> (BPUL) in Bioengineering NFL22, scale: 6 L, [https://2012.igem.org/Team:Bielefeld-Germany/Protocols/Materials#Autoinduction_medium autoinduction medium] + 60 µg/mL chloramphenicol, 37 °C, pH 7, agitation increased when pO<sub>2</sub> was below 30 %, OD<sub>600</sub> taken every hour. ]] |
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+ | <br style="clear: both" /> | ||
+ | [[File:Bielefeld2012_BPUL6LChromatogramm.jpg|600px|thumb|center|Figure:4]] | ||
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Revision as of 12:25, 23 September 2012
Summary
BPUL - Laccase from Bacillus pumilus DSM 27 (ATCC7061)
After the measurement of activity of BPUL we made a scale-up and fermented E. coli KRX with <partinfo>BBa_K863000</partinfo> in Braun Biostat B with a total volume of 3 L. Agitation speed, pO2 and OD600 were determined and illustrated in the following graph. We got a long lag phase of 2 hours due to a relativly old preculture. The cell growth caused a decrease in pO2 and after 3 hours the value fell below 50 %, so that the agitation speed was increased automatically. After 8,5 hours the deceleration phase started and therefore the agitation speed was descreased. The maximal OD600 of 3,53 was reached after 10 hours. The cells were harvested after 11 hours.
ECOL - Laccase from Escherichia coli BL21 (DE3)
XCCL - Laccase from Xanthomonas campestris pv. campestris B100
BHAL - Laccase from Bacillus halodurans C-125
TTHL - Laccase from Thermus thermophilus HB27
PCIL - Laccase from Pycnoporus cinnabarinus
TVEL5 - Laccase from Trametes versicolor
TVEL10 - Laccase from Trametes versicolor
TVEL13 - Laccase from Trametes versicolor
TVEL20 - Laccase from Trametes versicolor
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