Bio-Process Kinetics

1. Bio-Process Kinetics Process Biotechnology Anondho WIJANARKO University of Indonesia Simplified Lecture Notes For Instructor

2. To Dwi ImantiandAdam Aulia, Musa Maulana, Isa Dzulqarnain and Lovely Muhammad Al Muhammad

3. CONTENTS • Microbial Growth • Microbial Cell Growth • Classical & Empirical Growth Kinetics • Environmental Alteration Studies • Light Illumination Effect • Temperature Effect • Microbial Kinetic Studies • Non Elementer Reaction • Microbial Growth Reaction Kinetics

4. Microbial Growth

5. Microbial Cell Growth • Mode of Growth • Selective assimilation of nutrients and convert into and also include Chemical rearrangement of protoplasmic material characteristic of the particular organism • Production of an increased amount of nuclear substance and cell division

6. Growth Phase V IV X [g/l] I II III t [h]

7. Growth Phase • Induction Phase (Lag Phase) • Transient Phase (Acceleration Phase) • Exponential Phase • Stationary Phase (Declining Phase) • Death Phase

8. Question Sheet • Why microbial growth have an lag phase? • Why death phase could be occurred in microbial growth? • What is essential nutrient for growth of organism especially prokaryotes? • What is important factor for cell division?

9. Growth Approximation X [g/l] mf f 1.0 t X m m X i t [h]

10. Growth Constants Exponential – Stationary Growth Phase Total Biomass Production(G) Incident growth rate, Incident mean division rate (mf) Specific growth rate, Beginning mean division rate (m) Doubling time of population (tD); exponential growth phase

11. Classical Growth KineticsEmpirical Approximation • Monod Growth Kinetic • Tessier Growth Kinetic • Moser Growth Kinetic • Contois Growth Kinetic

12. Empirical Growth KineticsMedium constituent Inhibition • Andrews Growth Kinetic • Aiba Growth Kinetic

13. Growth KineticsMultiple essential nutrient • Bailey Growth Kinetic

14. Home Work • Which kinetic approximation do you choose in case of microbial growth of Hepatotoxin produced Oscilatoria Agardhii NIVA CYA 97 in low temperature? • Which empirical equation that you choose of inoculation of microorganism in case of multiple content limitation of nutrients, such as Mg2+, phosphate, Nitrate and organic compound? • Which kinetic approximation do you choose of cultivation photosynthetic microorganism that did not grew up in pH above 7.8?

15. Simple Bio-Production Kinetic • Cellular growth rate • Monod approximation • Yield factor • Substrate Utilization • Product Formation (Beginning of Stationary Phase)

16. Environmental Alteration Studies

17. Microbial Growth KineticEnviromental Condition • Direct Effects • Light Illumination (Energy Source) • Temperature • Essential nutrients content • Indirect Effects • Gas inlet volumetric rate • Gas inlet content • Liquid circulation rate • Non essential nutrients content

18. Light Illumination Effect • Oscillatoria agardhi Gomont • (Post AF, R de Witt, LC Mur, J. Plank. Res., 7 (1985) 487-495) [Chl a/X] m 288K 293K 0.010 290K 293K 0.2/h 283K 288K I 283K 20W/m 2 I 2 20W/m

19. Temperature effect • Modified Arhenius Model m Arhenius Approximation Microbial growth 1/T

20. Temperature EffectClassification of Microorganism Thermophiles m Mesophiles Psychrophiles Obligate Facultative T 273 293 313

21. Question Sheet • What is happen if microorganism is at 90oC? Why? • In case of decreasing of temperature about 20oC from optimum temperature, what is happen in case of microbial growth rate? • In case of ethanol production that was S. sake have ethanol tolerance around 10%, what do you do to make an whisky industry? • Why a shade microbe does not grew well in high light illumination and commonly have not high temperature resistance?

22. Temperature EffectCellular Consideration DBI • Psychrophile • Obligate • Protococcus Agardh SS 100-3 • Oscillatoria redekei Van Goor • Oscillatoria sp. SS 100-5 • Facultative • Anabena cylindrica Lemmerman • Oscillatoria Agardhi Gomont • Nostoc commune Antartica • Mesophile • Synechococcus leopoliensis • Anabaena variabilis IAM M3 • Microcystis Aeruginosa IAM M228 • Thermophile • Mastigocladus laminosus HTF • Synechococcus lividus OH75S • Synechcocus elongatus It 7S Obligate Psychrophiles 2.0 Facultative Mesophiles 1.0 Thermophiles GC content 0.3 0.5 0.7

23. Home Work • Why optimum specific growth rate values of psychrophile factually, lower than thermophile? • Why GC content of microbial DNA is important for classification of organism in terms of growth rate dependence on temperature? • What is DBI?

24. Microbial Kinetic Studies

25. Non Elementer Reaction • Common reaction rate • N = integer Elementer • N = non integer Non Elementer Non Elementer Example :

26. Reaction Mechanism Mechanism path is microscopic description of a chemical reaction that was composed in term of elementer reactions Chemical reaction Mechanism Path Fast reaction Slow reaction, DECISIVE Path Fast reaction Intermediete species

27. Question Sheet • What is mechanism path? • What definition of intermediate species? • What was become determining factor of reaction rate?

28. Microbial GrowthEnzymatic Reaction/Kinetic consideration • Michaelis-Menten Kinetics • Reaction mechanism • Kinetic equation • Substrate Actvation and Inhibition • Reaction mechanism • Kinetic equation • Product Activation and Inhibition • Reaction mechanism • Kinetic equation

29. Michaelis-Menten Kinetics • Reaction mechanism • Kinetic derivation

30. Home Work • Please exhibit kinetic derivation of substrate activation and inhibition? • Please exhibit kinetic derivation of product activation and inhibition? • What do you think about reaction kinetic if Km is high that was indicated in bioremediation of toluene by C. nivalis?

31. Literature • Aruga Y., Ecological Studies of Photosynthesis and Matter Production of Phytoplankton I: Seasonal Changes in Photosynthesis of Natural Phytoplankton, Bot. Mag. Tokyo, 78 (1965) 280-288 • Bailey JE and DF Ollis, Biochemical Engineering Fundamentals, McGraw-Hill Book Co., New York, 1986 • Boney AD, Phytoplankton, Edward Arnold Publ. Ltd., London, 1975 • Contois DE, Kinetics of Bacterial Growth: Relationship between Population Density and Specific Growth Rate of Continuous Cultures, J. Gen. Microbiol., 21 (1959) 40-50 • Castenholz RW, Laboratory Cultures of Thermophilic Cyanophyte, Symposium uber Cyanophytensystematik, 32 (1970) 538551 • Foy RH, CE Gibson and RV Smith, The Influence of Day-length, Light Intensity and Temperature, on the Growth of Planktonic Blue-Green Algae, J Phycol., 11 (1976) 151-163 • Frohlich BT, IA Webster, MM Ataai and ML Shuler, Photobioreactors: Model of Interaction of Light Intensity, Reactor Design and Algal Physiology, Biotechnol. Bioeng. Symp., 13 (1983) 331-350

32. 8. Leopold AC and PE Kriedmann, Plant Growth and Development, Tata McGraw-Hill Publ. Co., New Delhi, 1975 9. Pilling MJ and PW Seakins, Reaction Kinetics, Oxford Sc. Publ., Tokyo 10. Mosser JL and TD Brock, Temperature Optima for Algae Inhabiting Cold Mountains Stream, Arctic and Alpine Research, 8 (1976) 111-114 11. Murata N, Low Temperature effects on Cyanobacterial Membranes, J. Bioenerg. Biomembr., 21 (1989) 61-75 12. Post AF, R de Witt and LR Muur, Interaction between Temperature and Light Intensity on Growth and Photosynthesis of the Cyanobacterium Oscilatoria agardhii, J. Plankton Research, 7 (1985) 487-495 13. Sato N, N Murata, Y Miura and N Ueta, Effect of Growth Temperature on Lipid and Fatty Acid Composition in the Blue Green Algae, Anabaena variabilis and Anacystis nidulans, Biochim. Biophys. Act., 572 (1979) 19-28 14. Seaburg KG, BC Parker, RA Wharton Jr. and GM Simmons Jr., Temperature-Growth Responses of Algal isolates from Antartica Oases, J. Phycol., 17 (1981) 353-360