User:Fermenter User/Notebook/MACG F6

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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 F6

Day -1 (Apr 20 Mon)

  <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>                 4/20/2009 (10:20)  Prepare small culture for inoculum (4mL)
   <Sung-Hye>            4/20/2009 (11:00)  Pour media
                         4/20/2009 (11:00)  pH calibration
                                           No Antifoam added
                         4/20/2009 (12:07)  Autoclave bioreactor
                                            3 empty bottles/fermenter
                                            Base, Carbon source+Inoculation, Feeding

4/20/2009 (13:30) Cooling & Start Temp Control 4/20/2009 (14:15) Add carbon source using pumps 4/20/2009 (14:30) Connect Base (400 ml) Prime the pumps 4/20/2009 (14:35) Connect MeOH sensor 4/20/2009 (14:35) Calibrate dO2 4/20/2009 (14:35) Set dO2

4/20/2009 (14:35) Wrap bioreactors with foil 4/20/2009 (14:37) Start F1 software 4/20/2009 (14:38) Start F2 software 4/20/2009 (14:39) Start F3 software 4/20/2009 (14:45) Start pH control
   <Ann>                 4/20/2009 (15:00)  Prepare small culture for inoculum (10mL)
   <Ann>                 4/20/2009 (19:15)  Prepare small culture for inoculum (175mL)

Day 0 (Apr 21 Tue)

   <Ann>                4/21/2009 (09:00)   Fermentation 00:00 
                             Inoculation 50 ml each using pump.
   <Ann>                4/21/2009 (09:00)  Fermentation 00:00 
                             F1: OD (0.16) Contamination? (N)
                             F2: OD (0.16) Contamination? (N)
                             F3: OD (0.16) Contamination? (N)
   <Ann>                4/21/2009 (09:00)  Fermentation 00:00  
                             F1: 400 ml 
                             F2: 370 ml
                             F3: 370 ml
   <Ann>                4/21/2009 (14:00)  Fermentation 05:00 
                             F1: OD (0.59) Contamination? (N)
                             F2: OD (0.55) Contamination? (N)
                             F3: OD (0.57) Contamination? (N)

Day 1 (Apr 22 Wed)

   <SungHye>            4/22/2009 (07:30)  Fermentation 12:30  
                             F1: 320 ml 
                             F2: 290 ml
                             F3: 300 ml
                             Autoclaved meeding bottles
   <Ann>                4/22/2009 (09:00)  Fermentation 24:00 
                             F1: OD (7.7) Contamination? (N)
                             F2: OD (7.5) Contamination? (N)
                             F3: OD (7.6) Contamination? (N)
   <SungHye>            4/22/2009 (09:00)  Fermentation 24:00 
                             F1: Change Temp to 15C
                             F2: Change Temp to 22C                              
   <Ann>                4/22/2009 (09:00)  Fermentation 24:00 Induction 00:00 
                             F1: Induce with Galactose (55 mL)  
                                 Start feeding: (10 rpm)
                             F2: Induce with Galactose (55 mL)                                     
                                 Start feeding: (10 rpm)
                             F3: Induced with Galactose (130 mL, diluted x2.4)  
                                 Start feeding: (1 rpm with 32 rpm pump)                              
Flowate calibration of Watson pump for feeding. Click it to see large
Flowate calibration of Watson pump for feeding. Click it to see large
Flowate calibration of Watson pump (32 rpm) for feeding. Click it to see large
Flowate calibration of Watson pump (32 rpm) for feeding. Click it to see large


   <Ann>                4/22/2009 (11:00)  Fermentation 26:00 
                             F1: OD (8.1) Contamination? (N)F1: Induction 02:00 
                             F2: OD (7.4) Contamination? (N)F2: Induction 02:00 
                             F3: OD (7.5) Contamination? (N)F3: Induction 02:00 
   <Ann>                4/22/2009 (13:00)  Fermentation 28:00 
                             F1: OD (9.1) Contamination? (N)F1: Induction 04:00 
                             F2: OD (8.0) Contamination? (N)F2: Induction 04:00 
                             F3: OD (7.9) Contamination? (N)F3: Induction 04:00 
   <Ann>                4/22/2009 (15:00)  Fermentation 30:00 
                             F1: OD (8.7) Contamination? (N)F1: Induction 06:00 
                             F2: OD (9.0) Contamination? (N)F2: Induction 06:00 
                             F3: OD (8.2) Contamination? (N)F3: Induction 06:00 
   <Ann>                4/22/2009 (17:00)  Fermentation 32:00 
                             F1: OD (8.8) Contamination? (N)F1: Induction 08:00 
                             F2: OD (8.6) Contamination? (N)F2: Induction 08:00 
                             F3: OD (8.4) Contamination? (N)F3: Induction 08:00 
   <Ann>                4/22/2009 (19:00)  Fermentation 34:00 
                             F1: OD (8.7) Contamination? (N)F1: Induction 10:00 
                             F2: OD (8.5) Contamination? (N)F2: Induction 10:00 
                             F3: OD (9.1) Contamination? (N)F3: Induction 10:00 
   <Ann>                4/22/2009 (21:00)  Fermentation 36:00 
                             F1: OD (11.4) Contamination? (N)F1: Induction 12:00 
                             F2: OD (9.7) Contamination? (N)F2: Induction 12:00 
                             F3: OD (8.8) Contamination? (N)F3: Induction 12:00 

Day 2 (Apr 23 Thu)

   <Ann>                4/23/2009 (00:00)  Fermentation 39:00 
                             F1: OD (8.8) Contamination? (N)F1: Induction 15:00 
                             F2: OD (8.4) Contamination? (N)F2: Induction 15:00 
                             F3: OD (7.8) Contamination? (N)F3: Induction 15:00 
   <Ann>                4/23/2009 (00:00)  Fermentation 39:00 
                             F1: OD (10.3) Contamination? (N)F1: Induction 15:00 
                             F2: OD (9.1) Contamination? (N)F2: Induction 15:00 
                             F3: OD (7.8) Contamination? (N)F3: Induction 15:00 

Day 3 (Apr 24 Fri)

   <Ann>                4/24/2009 (00:00)  Fermentation 00:00 
                             F1: OD (10.3) Contamination? (N)F1: Induction 15:00 
                             F2: OD (9.1) Contamination? (N)F2: Induction 15:00 
                             F3: OD (7.8) Contamination? (N)F3: Induction 15:00 
   <SungHye>            4/24/2009 (00:00) 
                             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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