User:Fermenter User/Notebook/MACG F8R

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Background color codes:
Pastel Green: Major change of setup this time
yellow: Sung-Hye's correction or question to user

Letter color codes:
Black: Basic comment
Red: Emergency Information (Power shutdown, building access, Manager's absence etc)
Blue: Fermentation Setup information
Green: Discussion
Orange: Fermentation hrs
Purple: Induction hrs


MACG F8R

Day -1 (May 19 Tue)

  <Bioreactor headplate configuation>
   pH sensor: monitor or control pH
   dO2 sensor: monitor or control dO2
   Condenser: Gas outlet
   OD probe: monitor cell growth ==> Not used! Replate it with septum
   Triple: Addition of media, Acid or Base if necessary
   MeOH sensor: Monitor EtOH (byproduct)
   
   <Sung-Hye's Note>
    MeOH sensors also detects ethanol! :See reference #1 on the bottom of the page
   
   <Ann>                 5/12/2009 (11:10)  Prepare small culture for inoculum (7.5mL)
   <Ann/Sung-Hye>        5/19/2009 (10:30)  pH calibration
                                            No Antifoam added
                         5/19/2009 (13:00)  Pour media
                         5/19/2009 (13:30)  Autoclave bioreactor
                                            3 empty bottles/fermenter
                                            Base, Carbon source+Inoculation, Feeding
                         

5/19/2009 (15:30) Cooling & Start Temp Control 5/19/2009 (15:45) Start F1 software 5/19/2009 (15:46) Start F2 software 5/19/2009 (15:47) Start F3 software 5/19/2009 (15:50) Connect MeOH sensor 5/19/2009 (16:00) Connect Base (400 ml) Prime the pumps Set pH 5/19/2009 (16:30) Wrap bioreactors with foil


   <Ann>                 5/12/2009 (02:50)  Prepare small culture for inoculum (15mL)
   <Ann>                 5/12/2009 (17:30)  Prepare small culture for inoculum (315mL)

Day 0 (May 20 Wed)

   <Ann/SungHye>         5/20/2009 (07:10)  Add carbon source using pumps 
                         5/20/2009 (07:20)  Calibrate dO2
                         5/20/2009 (07:20)  Set dO2

   <Ann>                 5/20/2009 (08:00)   Fermentation 00:00 
                             Inoculation 50 ml (F1 and F2) and 250 ml (F3) using pump.
   <Ann>                 5/20/2009 (08:00)  Fermentation 00:00 
                             F1: OD (0.222) Contamination? (N)
                             F2: OD (0.132) Contamination? (N)
                             F3: OD (0.227) Contamination? (N)
   <Ann>                 5/20/2009 (12:00)  Fermentation 04:00 
                             F1: OD (0.484) Contamination? (N)
                             F2: OD (0.496) Contamination? (N)
                             F3: OD (0.466) Contamination? (N)
   <Ann>                 5/20/2009 (17:00)  Fermentation 09:00 
                             F1: OD (2.12) Contamination? (N)
                             F2: OD (1.89) Contamination? (N)
                             F3: OD (1.69) Contamination? (N)

Day 1 (May 21 Thu)

   <Ann>                 5/20/2009 (07:00)  Fermentation 23:00 
                             F1: OD (17.5) Contamination? (N)
                             F2: OD (10.9) Contamination? (N)
                             F3: OD (9.8) Contamination? (N)
   <Ann>                 5/20/2009 (07:00-08:30)  Fermentation 23:00-24:30 
                             F1: Spin down cells and re-suspend with fresh media (1-L)
                                 Pour culture back into fermenter
                             F2: Spin down cells and re-suspend with fresh media (1-L)
                                 Pour culture back into fermenter
                             F3: Spin down cells and re-suspend with fresh media (1-L)
                                 Pour culture back into fermenter
   <Ann>                 5/20/2009 (08:30)  Fermentation 24:30 
                             F1: OD (17.1) Contamination? (N)
                             F2: OD (9.2) Contamination? (N)
                             F3: OD (8.4) Contamination? (N)
   <Ann>                 5/20/2009 (08:30)  Fermentation 24:30 Induction 00:00 
                             F1: Induce with Galactose (55 mL)  
                             F2: Induce with Galactose (55 mL)                                     
                             F3: Induce with Galactose (55 mL)  
   <Ann>                 5/20/2009 (12:30)  Fermentation 28:30 
                             F1: OD (22.5) Contamination? (N)F1: Induction 04:00 
                             F2: OD (10.7) Contamination? (N)F2: Induction 04:00 
                             F3: OD (9.5) Contamination? (N)F3: Induction 04:00 
   <Ann>                 5/20/2009 (16:30)  Fermentation 32:30 
                             F1: OD (37.7) Contamination? (N)F1: Induction 08:00 
                             F2: OD (13.4) Contamination? (N)F2: Induction 08:00 
                             F3: OD (12.3) Contamination? (N)F3: Induction 08:00 
   <SungHye>             5/20/2009 (16:30)  Fermentation 32:30 
                             Harvest

References

  1. www.ravenbiotech.com/product.html [Paper1]


  1. High Cell Density Fermentation of Saccharomyces cerevisiae JUL3 in Fed-batch Culture for the Production of β-Glucan (2007) 13, 1, 153-158 [Paper2]
  Image:IE13-1-0153.pdf
  Fermentation condition: 30°C, 200 rpm, no pH control, 1 vvm
  
  1. Optimization of enterokinase fermentation using a recombinant Saccharomyces cerevisiae. (2005) 40, 717-722 [Paper3]
  Image:PB-40-717.pdf
  Fermentation condition: 30°C, 500 rpm, no pH control, 2 vvm
Error fetching PMID 16820461:
  1. Error fetching PMID 16820461: [Paper4]
  1. Peamiability test: Aliquots (100 µl) of this cell suspension were pipetted into fluoroplate wells, 
     which contained NPN (10 µM) and, as test substances, either EDTA (1.0 and 0.1 mM), PEI (10 µg ml–1), 
     DMSA (1 mM), AOT (1 mM), or HEPES buffer (control) to make up a total volume of 200 µl. If desired, 
     MgCl2 was added to the cell suspension before addition of NPN. Fluorescence was monitored within 
     3 min from four parallel wells per sample (excitation, 355 nm; half bandwidth, 38 ± 3 nm; emission, 
     402 nm; half bandwidth, 50 ± 5 nm). Each assay was performed at least three times. 
Error fetching PMID 9534237:
  1. Error fetching PMID 9534237: [Paper5]
  1. OD600 unit is equivalent to 0.64 +/- 0.02 g cells (dry weight)/L
Error fetching PMID 17111383:
  1. Error fetching PMID 17111383: [Paper6]
  1. To demonstrate the localization of the proteins relative to the cells, cell contours were visualized 
     with Alexa 647-concanavalin A.
Error fetching PMID 18451787:
  1. Error fetching PMID 18451787: [Paper7]
Error fetching PMID 17709746:
  1. Error fetching PMID 17709746: [Paper8]
Error fetching PMID 11005931:
  1. Error fetching PMID 11005931: [Paper8]

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