Team:Bielefeld-Germany/Results/Summary
From 2012.igem.org
(→BPUL - Laccase from Bacillus pumilus DSM 27 (ATCC7061)) |
(→ECOL - Laccase from Escherichia coli BL21 (DE3)) |
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- | [[File:Bielefeld2012_ECOL3LFermentation.jpg|600px|thumb|center|Fermentation of ''E. coli'' KRX with <partinfo> | + | [[File:Bielefeld2012_ECOL3LFermentation.jpg|600px|thumb|center|Fermentation of ''E. coli'' KRX with <partinfo>BBa_K863005</partinfo> (ECOL) in Infors Labfors Bioreactor, 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_ECOL3LChromatogramm.jpg|600px|thumb|center|Figure 2: Chromatogramm of Wash and Elution from FLPC Ni-NTA-Histag Purification of ECOL produced by 3 L fermentation of ''E. coli'' KRX with <partinfo> | + | [[File:Bielefeld2012_ECOL3LChromatogramm.jpg|600px|thumb|center|Figure 2: Chromatogramm of Wash and Elution from FLPC Ni-NTA-Histag Purification of ECOL produced by 3 L fermentation of ''E. coli'' KRX with <partinfo>BBa_K863005</partinfo>.]] |
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Revision as of 13:14, 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 Figure 1. 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.
Another scale-up of the fermentation of E. coli KRX with <partinfo>BBa_K863000</partinfo> was made up to a final working volume of 6 L in Bioengineering NFL22. Agitation speed, pO2 and OD600 were determined and illustrated in Figure 3. There was no noticeable lag phase.
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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