Design of Enzyme Reactor

							Design of Enzyme Reactor


     Geoff Greer
     Joseph Gvildys
     Aaron McGregor

     April 2nd, 2007
    Enzyme Selection
   Transpeptidation
       Trypsin or
        Achromobacter lyticus
        proteinase I
   Optimal enzyme
    conditions
       4oC and a pH of 7




                                Eli Lilly
Enzyme Immobilization

   Benefits
       Savings on enzyme requirements and
        downstream processing
   Trypsin-Agarose
       $211 for 100 units (~ 3mL)

               Property

           Biological Source     trypsin from bovine pancreas
           Extent of Labeling   15-35 units per ml (packed gel)
                Matrix                 beaded agarose
          Storage Temperature               2-8oC
Reactor Comparison

   Batch
       High operating costs
       Batch-to-batch variations
   PFR
       Difficult to control pH and Temperature
   CSTR
       Simple pH, Temperature control
       Simple catalyst charging and replacement
Design Parameters
     Parameter           Value
         Q              7.786 L/h
        [Ro]          0.0179 mol/L
        Vmax       2.35 × 10-7 mol/L.s
        Km         9.90 × 10-5 mol/L
        k2             0.1285 s-1
      MWreactant      6000 g/mol
      MWtrypsin      25 000 g/mol
Reactor Design

   Specify conversion, calculate [R]
   From [R], calculate v
                 Vm ax[ R ]
              v
                 K m  [ R]

   From v, calculate D
                     Vm ax[ R ]
         D
            ( K m  [ R ])([ Ro ]  [ R ])
Reactor Design

   From D, calculate V
           Q  D V
   From V, calculate tres
            t res  1 / D
   From Vmax, k2, calculate [Eo] and total
    amount of enzyme required
            E o  Vm ax / k 2
Reactor Sizing
Reactor Sizing

   Conversion of 99.25%
       120 L reactor required
   Increase conversion by 0.5% to 99.75%
       360 L reactor required
Enzyme required

   Required amount of enzyme is
    1.83 μmol/L
       5.5 g pure trypsin
       2.6 kg immobilized trypsin
           Based on difference in activity between
            immobilized and non-immobilized enzyme
           ~ $18 000
Reactor Design Considerations

   Temperature control
   pH control
   Level control
   Sterilizable
   Wire-mesh sieve
   Sample port
Simplified Reactor Schematic
Costing of the Reactor
   Several different manufacturers
       New Brunswick Scientific
       Novaferm AB
       Scigenics
       New MBR
   Quote from New Brunswick Scientific
       BioFlo® Pro – 150 L (120 L working volume)
           $140 000 (US)
       Batch reactor
Final Reactor Schematic
Overall Cost of Unit

   For reactor and enzyme, estimated
    cost is at least $160 000
       Additional considerations
           Reactor modifications
           De-naturation of enzyme
Thank You

   Questions?