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INDUSTRIAL ENZYMES FOR SUSTAINABLE BIO-ECONOMY:
Large scale production and application in industry, environment, and agriculture



                               Amare Gessesse
     Nelson Mandela African institute of Science and Technology (NM-AIST),
                               Arusha, Tanzania

           First Bio-Innovate Regional Scientific Conference
             Addis Ababa, Ethiopia, 25-27 February 2013
Participating laboratories
No. Academic Institution                                    Country
1    Nelson Mandela African Institute of Science and        Tanzania
     Technology (NM-AIST)
2    Addis Ababa University                                 Ethiopia
3    University of Nairobi                                  Kenya
4    University of Dar es Salaam                            Tanzania
5    National University of Rwanda                          Rwanda
6    Lund University                                        Sweden
7    Natnl Inst for Interdisciplinary Science and Technology India

                         Industrial partners
8    Leather Industry Development Institute                 Ethiopia
9    Modjo Tannery                                          Ethiopia
10   Sumbawanga Agriculture and Animal Food Industry        Tanzania
11   Bekas Chemicals Plc                                    Ethiopia
Major Aim


Use the region’s biodiversity as a source of
        Novel Industrial Enzymes!
Africa’s Biodiversity
Produsts from Our Microbial Biodiversity
                        > 73 C




  pH > 10
                        > 95 C
Industrial Enzymes



Industry                   Environment          Agriculture

      Leather tanning           Degradation        Animal feed
      Starch processing         of residual        additive
                                waste
      Detergent                                     Feed
                                Detoxificatio       processing
      Textile
                                n of toxic
      Chemical/                 substances         Agroprocessing
      Pharmaceutical
      Food processing

      Protein hydrolysis

      Biofuel
Advantages of using industrial
              enzymes?
           Direct                          Indirect

• Foreign currency               • Reduction in environmental
       • Export of enzymes         pollution
       • Value addition to raw          • Cleaner production
         materials

                                 • Help to save foreign
• Job creation                     currency
       • Enzyme production
                                        • Example: the leather industry
       • Value addition
Current Worldwide Industrial
        Enzyme Production
• Over 120 enzyme
  companies
      • Dominated by Europe and
        North America
      • Control >90% of the
        world industrial enzyme
        market
• None from SS Africa
                                  Europe and North America control
• Enormous potential in           >90% of world enzyme market

  SS Africa
                                   Why?
Africa’s Performance in Science & Technology




                        Population            Tertiary Education




                Scientific research     Patent registration

Source: www.worldmapper.org
Research on enzymes
Alkaline proteases
pH profile of AL-20 (A) and AL-89
proteases (B)                       Temperature profile of AL-20
                                    (A) and AL-89 proteases (B)
A rial
  S tability of A L-20 protease w ith and w ithout E D T A treatm ent
                                                                                                                                                                                                                                                100




                                                                                                                                                                                        R e s id u a l a c tiv ity (% )
                                                                     100                                                                                                                                                                                    80
          R e s id u a l a c tiv ity (% )




                                                                      80                                                                                                                                                                                    60

                                                                      60                                                                                                                                                                                    40
                                                                                         65°C
                                                                      40
                                                                                                                                                                                                                                                            20
                                                                      20
                                                                                                                                                                                                                                                             0
                                                                       0                                                                                                                                                                                         0       10    20    30      40    50     60    70
                                                                           0        50          100           150    200
                                                                                                                                                                                                                                                                                    Tim e (m in)
                                                                                          T im e (m in)                   E D T A treated and assayed w ith E D T A
                                                                                                                                                                      2+
                                                                                                                          N o E D T A and assayed w ith 10 m M C a



                                                                      100
                                                                                                                                                                             V ia
                                   R e s id u a l a c tiv ity (% )




                                                                                                          70°C                                                                                                                                              100




                                                                                                                                                                                                                          R e s id u a l a c tiv ity (% )
                                                                       80
                                                                                                                                                                                                                                                             80
                                                                       60

                                                                       40
                                                                                                                                                                                                                                                             60

                                                                       20                                                                                                                                                                                    40

                                                                           0                                                                                                                                                                                 20
                                                                               0   10    20      30     40      50   60
                                                                                                                                                                                                                                                                 0
                                                                                              T im e (m in)
                                                                                                                                                                                                                                                                     0    10    20      30    40     50    60    70

                                                                                                                                                                                                                                                                                      Tim e (m in)



S ta b ility o f A L -2 0 p ro te a se to E D T A tre a tm e n t                                                                                                           F ig.4. E ffect of detergents on stability of A L-20 protease and other proteases.
                                                                                                                                                                           A L-20 protease at 60°C (circle), subtilisin at 50°C (square), protenase K at 60°C
                                                                                                                                                                           (triangle), and endogenous enzym e at 50°C (diam ond)
Use of a new fungal neutral protease
       isolated at AAU for bating
Test                                            Unit      Comm.            Protease
                                                          enzyme           BACC 480

Tensile strength                                N/mm2     28.2             29.5
Percentage elongation                           %         59.4             68.1
Tear load                                       N/mm      39.7             37.7
Mean tear load (parallel to the back bone)      N         28.0             24.0
Mean tear load (perpendicular to the back)      N         28.0             29.0
Average tear load                               N         28.0             26.5
Distension at burst                             mm        11.8             11.2

       Tested at the Leather Industry Development Institute, Addis Ababa

        (Tested with staff from LIDI)
Alkaline protease from alkaliphiles
• Several alkaline                    (Buffer alone)      (Buffer + enzyme)

  protease producers
      • From soda lakes


• Example:                                             Sheep skin

      • Vibrio sp R-11 protease         (Buffer alone)      (Buffer + enzyme)
      • Interesting application for
        dehairing



                                                       Cow hide
Enzyme production potential
               benefits
• In enzyme production
      • Substrate account for 30 – 40%
        of the cost
      • Here we use hair & mineral salt
        solution
• Use of the enzyme lead to
  reduction in pollution
      • Na2S
      • Lime
      • Protein
• Reduction in import
      • Na2S
      • Other chemicals
Detergent applications
• Detergent industry            As detergent additive

      • Alkaline proteases
      • Alkaline amylases


• Protein hydrolysis
      • Microbiological media
      • Animal feed
      • Food application
• Enzyme production cost
      • Considered as key
Enzymes for processing of starch
         containing crops
• Starch sources
      • Cassava
      • Enset
      • Cereals
• Products
      •   Glucose
      •   Fructose
      •   Maltose
      •   Confectionary
      •   Jams
      •   Beverages
      •   etc
Animal feed
• Feed utilization by
  monogastric animals
     • Unable to digest
       pentosatans
     • Low availability of
       phosphate


• Enzymes
     • Xylanases
     • Phytase
Xylanase and cellulase activities of T.
   reesei Rut C-30 (Mulaa et al.)
Pulp and paper industry
Microbial
                             Strain
           Submerged                                      Solid state
          fermentation                                   fermentation

                                                                         Animal
                             Cell free                Extraction &
              Cell                                                        Feed
Biomass                    supernatant                 separation
           separation
                                                          Solvent
Compost
                                        Supernatant          Solvent
                                                                         Spent
                                                             recovery
                           Enzyme
                          Concentrate
                                                                        Biogas

                            Enzyme
                          Stabilization
                                                                        Compost

                         Enzyme product
Main considerations
• Nature of the organism         • Environmental issues
      • Growth under SSF or            • Reduction of pollution
        SmF                            • Serve as a model
      • Depend on the organism
                                 • Byproduct utilization
• Relative enzyme yield            for cost reduction
      • SSF                            • Feed
      • SmF                            • Biogas
• Cost                                 • Compost
      • Initial investment
      • Product recovery
      • Growth substrate
Scaling up fermentation process?
• Mode of fermentation
     • Solid state fermentation
     • Submerged fermentation
• Organisms
     • Bacteria
     • Fungi
• Enzymes
     • Proteases
     • Amylases
     • Xylanases, etc
Gap in the transfer of research results to
         commercial products
• The gap
      • How to scale up?
      • Absence of private sector
        (or is risk avert)
      • Lack of entrepreneurship
        skill in academia
      • Capital
      • Lack of role models


• Important to bridge the
  gap
Can we change course?
• Scale up production
      • Pilot scale
• Evaluation under
  application condition
      • Stabilization
• Collaboration with end
  users
• Ultimately
      • Can we establish new
        companies?
If yes, what do new companies need?
• Proper care
      • Public sector
      • Creation of an enabling
        environment


• Those in academia must
      • Learn entrepreneurship
        skills
      • Proactive
We need to jump big!
We need to jump big!




   Thank you!

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Industrial enzymes for sustainable bio-economy: Large scale production and application in industry, environment, and agriculture

  • 1. INDUSTRIAL ENZYMES FOR SUSTAINABLE BIO-ECONOMY: Large scale production and application in industry, environment, and agriculture Amare Gessesse Nelson Mandela African institute of Science and Technology (NM-AIST), Arusha, Tanzania First Bio-Innovate Regional Scientific Conference Addis Ababa, Ethiopia, 25-27 February 2013
  • 2. Participating laboratories No. Academic Institution Country 1 Nelson Mandela African Institute of Science and Tanzania Technology (NM-AIST) 2 Addis Ababa University Ethiopia 3 University of Nairobi Kenya 4 University of Dar es Salaam Tanzania 5 National University of Rwanda Rwanda 6 Lund University Sweden 7 Natnl Inst for Interdisciplinary Science and Technology India Industrial partners 8 Leather Industry Development Institute Ethiopia 9 Modjo Tannery Ethiopia 10 Sumbawanga Agriculture and Animal Food Industry Tanzania 11 Bekas Chemicals Plc Ethiopia
  • 3. Major Aim Use the region’s biodiversity as a source of Novel Industrial Enzymes!
  • 5. Produsts from Our Microbial Biodiversity > 73 C pH > 10 > 95 C
  • 6. Industrial Enzymes Industry Environment Agriculture Leather tanning Degradation Animal feed Starch processing of residual additive waste Detergent Feed Detoxificatio processing Textile n of toxic Chemical/ substances Agroprocessing Pharmaceutical Food processing Protein hydrolysis Biofuel
  • 7. Advantages of using industrial enzymes? Direct Indirect • Foreign currency • Reduction in environmental • Export of enzymes pollution • Value addition to raw • Cleaner production materials • Help to save foreign • Job creation currency • Enzyme production • Example: the leather industry • Value addition
  • 8. Current Worldwide Industrial Enzyme Production • Over 120 enzyme companies • Dominated by Europe and North America • Control >90% of the world industrial enzyme market • None from SS Africa Europe and North America control • Enormous potential in >90% of world enzyme market SS Africa Why?
  • 9. Africa’s Performance in Science & Technology Population Tertiary Education Scientific research Patent registration Source: www.worldmapper.org
  • 12. pH profile of AL-20 (A) and AL-89 proteases (B) Temperature profile of AL-20 (A) and AL-89 proteases (B)
  • 13. A rial S tability of A L-20 protease w ith and w ithout E D T A treatm ent 100 R e s id u a l a c tiv ity (% ) 100 80 R e s id u a l a c tiv ity (% ) 80 60 60 40 65°C 40 20 20 0 0 0 10 20 30 40 50 60 70 0 50 100 150 200 Tim e (m in) T im e (m in) E D T A treated and assayed w ith E D T A 2+ N o E D T A and assayed w ith 10 m M C a 100 V ia R e s id u a l a c tiv ity (% ) 70°C 100 R e s id u a l a c tiv ity (% ) 80 80 60 40 60 20 40 0 20 0 10 20 30 40 50 60 0 T im e (m in) 0 10 20 30 40 50 60 70 Tim e (m in) S ta b ility o f A L -2 0 p ro te a se to E D T A tre a tm e n t F ig.4. E ffect of detergents on stability of A L-20 protease and other proteases. A L-20 protease at 60°C (circle), subtilisin at 50°C (square), protenase K at 60°C (triangle), and endogenous enzym e at 50°C (diam ond)
  • 14.
  • 15. Use of a new fungal neutral protease isolated at AAU for bating Test Unit Comm. Protease enzyme BACC 480 Tensile strength N/mm2 28.2 29.5 Percentage elongation % 59.4 68.1 Tear load N/mm 39.7 37.7 Mean tear load (parallel to the back bone) N 28.0 24.0 Mean tear load (perpendicular to the back) N 28.0 29.0 Average tear load N 28.0 26.5 Distension at burst mm 11.8 11.2 Tested at the Leather Industry Development Institute, Addis Ababa (Tested with staff from LIDI)
  • 16. Alkaline protease from alkaliphiles • Several alkaline (Buffer alone) (Buffer + enzyme) protease producers • From soda lakes • Example: Sheep skin • Vibrio sp R-11 protease (Buffer alone) (Buffer + enzyme) • Interesting application for dehairing Cow hide
  • 17. Enzyme production potential benefits • In enzyme production • Substrate account for 30 – 40% of the cost • Here we use hair & mineral salt solution • Use of the enzyme lead to reduction in pollution • Na2S • Lime • Protein • Reduction in import • Na2S • Other chemicals
  • 18. Detergent applications • Detergent industry As detergent additive • Alkaline proteases • Alkaline amylases • Protein hydrolysis • Microbiological media • Animal feed • Food application • Enzyme production cost • Considered as key
  • 19. Enzymes for processing of starch containing crops • Starch sources • Cassava • Enset • Cereals • Products • Glucose • Fructose • Maltose • Confectionary • Jams • Beverages • etc
  • 20.
  • 21.
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  • 23. Animal feed • Feed utilization by monogastric animals • Unable to digest pentosatans • Low availability of phosphate • Enzymes • Xylanases • Phytase
  • 24. Xylanase and cellulase activities of T. reesei Rut C-30 (Mulaa et al.)
  • 25. Pulp and paper industry
  • 26. Microbial Strain Submerged Solid state fermentation fermentation Animal Cell free Extraction & Cell Feed Biomass supernatant separation separation Solvent Compost Supernatant Solvent Spent recovery Enzyme Concentrate Biogas Enzyme Stabilization Compost Enzyme product
  • 27. Main considerations • Nature of the organism • Environmental issues • Growth under SSF or • Reduction of pollution SmF • Serve as a model • Depend on the organism • Byproduct utilization • Relative enzyme yield for cost reduction • SSF • Feed • SmF • Biogas • Cost • Compost • Initial investment • Product recovery • Growth substrate
  • 28. Scaling up fermentation process? • Mode of fermentation • Solid state fermentation • Submerged fermentation • Organisms • Bacteria • Fungi • Enzymes • Proteases • Amylases • Xylanases, etc
  • 29. Gap in the transfer of research results to commercial products • The gap • How to scale up? • Absence of private sector (or is risk avert) • Lack of entrepreneurship skill in academia • Capital • Lack of role models • Important to bridge the gap
  • 30. Can we change course? • Scale up production • Pilot scale • Evaluation under application condition • Stabilization • Collaboration with end users • Ultimately • Can we establish new companies?
  • 31. If yes, what do new companies need? • Proper care • Public sector • Creation of an enabling environment • Those in academia must • Learn entrepreneurship skills • Proactive
  • 32. We need to jump big!
  • 33. We need to jump big! Thank you!