User:Fermenter User/Notebook/MACG F1

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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 F1 (F3)

Day -2 (Feb 8 Sun)

   <Ann>                 2/8/2009 (17:30) Prepare for small culture. (15mL culture from plate)

Day -1 (Feb 9 Mon)

  <Bioreactor headplate configuation>
   pH sensor: monitor or control pH
   dO2 sensor: monitor or control dO2
   Condenser: Gas outlet
   OD probe: monitor cell growth
   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
   
 
   <Sung-Hye>            2/9/2009 (9:40)  Pour media
                         2/9/2009 (9:46)  Antifoam added
                         2/9/2009 (9:57)  pH calibration
                         2/9/2009 (10:14) Autoclave bioreactor
                                           2 empty bottles/fermenter
                                           Carbon source+Inoculation

==> Autoclave malfuntion? All media came out of bioreactor

2/9/2009 (12:23) Make media again: 3.4-L (380 ml x10 + Water) 2/9/2009 (12:23) Antifoam added 2/9/2009 (12:25) pH calibration 2/9/2009 (12:40) Autoclave bioreactor 2/9/2009 (14:14) Start Temp Control 2/9/2009 (14:18) Start add rest of media and carbon sources using pumps at Temp 55°C 3.4-L media (autoclave) 1.1-L media 0.25-L Carbon Total 4.75 2/9/2009 (14:41) Start OD software: ended 2/9/2009 (14:44) Start F3 software 2/9/2009 (15:41) Restart OD software
   <Ann>                 2/9/2009 (14:05) Prepare small culture for inoculum. (350mL)

Day 0 (Feb 10 Tue)

   
    Sung-Hye: CHBE Class Auditing 9:30 - 11:00
   
   <Sung-Hye>           2/10/2009 (7:30)  Calibrate dO2
                        2/10/2009 (8:10)  Set dO2
   <Sung-Hye and Ann>   2/10/2009 (9:00)   Fermentation 0:00 
                             Inoculation 250 ml each using pump.
   <Ann>                2/10/2009 (11:00)  Fermentation 2:00 
                             F3: OD (0.465) Contamination? (N)
   <Ann>                2/10/2009 (13:30)  Fermentation 04:30 
                             F3: OD (0.600) Contamination? (N)
   <Ann>                2/10/2009 (15:30)  Fermentation 06:30 
                             F3: OD (0.860) Contamination? (N)
   <Ann>                2/10/2009 (17:00)  Fermentation 08:00 
                             F3: OD (0.980) Contamination? (N)
   <Ann>                2/10/2009 (17:30)  Fermentation 08:30 
                             F3: OD (1.020) Contamination? (N)

Day 1 (Feb 11 Wed)

   <Ann>                2/11/2009 (9:00)   Fermentation 24:00  Induction 00:00 
                             F3: OD (5.98) Contamination? (N) 
                             Start feeding: 2.7 ml/hr
File:Watsonpump.tif
   <Ann>                2/11/2009 (13:00)   Fermentation 28:00 
                             F3: OD (5.90) Contamination? (N)  Induction 04:00 
   <Ann>                2/11/2009 (17:00)   Fermentation 32:00 
                             F3: OD (8.42) Contamination? (N)  Induction 08:00 

Day 2 (Feb 12 Thu)

   <Ann>                2/12/2009 (01:00)   Fermentation 40:00 
                             F3: OD (7.28) Contamination? (N)  Induction 16:00 
   <Ann>                2/12/2009 (07:00)   Fermentation 46:00 
                             F3: OD (8.48) Contamination? (N)  Induction 22:00 
   <Sung-Hye and Ann>   2/12/2009 (9:00)    Fermentation 49:00 
                            Added rest of feed solution (~200mL) Induction 25:00 
   <Ann>                2/12/2009 (14:00)   Fermentation 53:00 
                             F3: OD (9.12) Contamination? (N)  Induction 29:00 
   <Sung-Hye>           2/12/2009 (16:00)  Fermentation 55:00 Induction 31:00
                             Calibrate EtOH
   <Ann>                2/12/2009 (16:00)  Fermentation 55:00 Induction 31:00
                             Possible 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]
  File: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]
  File:PB-40-717.pdf
  Fermentation condition: 30°C, 500 rpm, no pH control, 2 vvm
  1. Alakomi HL, Paananen A, Suihko ML, Helander IM, and Saarela M. Weakening effect of cell permeabilizers on gram-negative bacteria causing biodeterioration. Appl Environ Microbiol. 2006 Jul;72(7):4695-703. DOI:10.1128/AEM.00142-06 | PubMed ID:16820461 | HubMed [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. 
  1. Hounsa CG, Brandt EV, Thevelein J, Hohmann S, and Prior BA. Role of trehalose in survival of Saccharomyces cerevisiae under osmotic stress. Microbiology. 1998 Mar;144 ( Pt 3):671-80. DOI:10.1099/00221287-144-3-671 | PubMed ID:9534237 | HubMed [Paper5]
  1 OD600 unit is equivalent to 0.64 +/- 0.02 g cells (dry weight)/L
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