ERT 416/3 CHAPTER 10 :

1. ERT 416/3CHAPTER 10: INTEGRATED BIOSEPARATION SCHEME FOR PRODUCT ISOLATION, PURIFICATION AND FORMULATIONS UNITS FOR BIOPROCESS MISS. RAHIMAH BINTI OTHMAN (Email: rahimah@unimap.edu.my)

2. COURSE OUTCOME 1 CO1) CHAPTER 7 : Upstream Processing In Bioprocess Plant. CHAPTER 8 : Bioreaction Design In Bioprocess Plant. CHAPTER 9 : Downstream Processing In Bioprocess Plant. CHAPTER 10: Integrated Bioseparation Scheme for Product Isolation, Purification and Formulation Units For Bioprocess. CHOOSE the appropriate sequencing of processes; SYNTHESIZEand EVALUATE integrated bio-separation schemes that could be used to achieved the required purification of important biological molecules (product).

3. What is the “Integration Process”? What is “Product Isolation, Purification and Formulation Units”. How to CHOOSE the appropriate sequencing of processes. SYNTHESIZEand EVALUATE integrated bio-separation schemes. - [Propose: to achieve the required purification of important biological molecules (product)]. OUTLINES

4. GENERALIZED VIEW OF BIOPROCESS

5. WHAT IS INTEGRATION PROCESS? INTEGRATION In engineering, system integration is the bringing together of the component subsystems into one system and ensuring that the subsystems function together as a system. In information technology, systems integration is the process of linking together different computing systems and software applications physically or functionally.

6. EXAMPLE: The research roadmap of Process Integration study in Yeast & Ethyl Alcohol (Y&EA) plant.

7. Physical model of Y&EA plant.

8. Fig. 5. Scheme of the integrated bioprocess. Limonene-saturated air was supplied to the bioreactor by leading it through a washing flask filled with pure limonene (A).Glycerol (B) and limonene (C) were continuously fed via peristaltic pumps. Using another peristaltic pump (D) the culture broth was continuously recirculated through a recovery loop containing a column with a fluidized bed of anion exchanger Amberlite IRA 410 Cl (E).

9. CHEMICAL MOLECULE STRUCTURE INTEGRATION

10. What is “Product Isolation, Purification, and Formulation Units? Removal of insolubles is the first step and involves the capture of the product as a solute in a particulate-free liquid, for example the separation of cells, cell debris or other particulate matter from fermentation broth containing an antibiotic. - Typical operations to achieve this are filtration, centrifugation, sedimentation, flocculation, electro-precipitation, and gravity settling. - Additional operations such as grinding, homogenization, or leaching, required to recover products from solid sources such as plant and animal tissues, are usually included in this group. Product Isolation is the removal of those components whose properties vary markedly from that of the desired product. - For most products, water is the chief impurity and isolation steps are designed to remove most of it, reducing the volume of material to be handled and concentrating the product. - Solvent extraction, adsorption, ultrafiltration, and precipitation are some of the unit operations involved.

11. What is “Product Isolation, Purification, and Formulation Units? Product Purification is done to separate those contaminants that resemble the product very closely in physical and chemical properties. - Consequently steps in this stage are expensive to carry out and require sensitive and sophisticated equipment. - This stage contributes a significant fraction of the entire downstream processing expenditure. - Examples of operations include affinity, size exclusion, reversed phase chromatography, crystallization and fractional precipitation. Product Polishing describes the final processing steps which end with packaging of the product in a form that is stable, easily transportable and convenient. - Crystallization, desiccation, lyophilization and spray drying are typical unit operations.

12. How to CHOOSE the appropriate sequencing of processes? Process Integration (PI), is primary focused to system oriented process design for energy efficient and cost effective mapping of the plant. This approach can be summarised in one sentence: ‘‘PI is probably the best approach that can be used to obtain significant energy and water savings as well as pollution reductions for different kind of industries’’.

13. How to CHOOSE the appropriate sequencing of processes? Integrated bioprocesses have been developed to optimise yield and cost-effectiveness of production of low and high molecular weight molecules. Most of the recent research describes the integration of processes, especially the ISPR (in situ product removal) technique, during the production of low molecular weight compounds. To achieve a continuous product recovery (i.e. ISPR), techniques based on extraction (i.e. the most commonly used method), adsorption, electrodialysis and crystallisation are used. Other integrated processes include integrated product formation in which the breakdown product of a polymer is converted to a low molecular compound in a one-pot process, and integrated extraction and enzymatic conversion.

15. How to CHOOSE the appropriate sequencing of processes? In situ product removal (ISPR) • I n situ extraction of other systems • In situ adsorption and ion exchange • In situ electrodialysis • In situ crystallisation Integrated product formation EXAMPLE: The decomposition of a polymer (starch, cellulose, hemicellulose, protein) and the conversion of the monomer are performed sometimes in a one-pot process. Such a process is the simultaneous saccharification of corncobs by the enzyme of Trichoderma reesei and the conversion of the product by L.rhamnosus to lactic acid .

16. How to CHOOSE the appropriate sequencing of processes? • EXAMPLE:Membrane-based separations

17. SYNTHESIZE and EVALUATE integrated bio-separation schemes. Integrated stack-monitoring emission system.

18. Detailed flowsheet of Yeast & Ethyl Alcohol Plant.

24. References and recommended reading Freeman A, Woodley J, Lilly MD: In situ product removal as a tool for bioprocessing. Biotechnology 1993, 11:1007-1012. Schuegerl K: Solvent Extraction in Biotechnology. Springer Verlag; 1994. Schuegerl K, Kretzmer G, Freitag R, Scheper T: Integrierte biotechnologische produktionsprozesse. Chem Ing Technik 1994, 66:1585-1592. [Title translation: Integrated biotechnological processes.] 4. Schuegerl K: Integrated processing of biotechnology products. Biotechnol Adv 2000, 18:581-599. Fernandes P, Prazeres DM, Cabral JMS: Membrane assisted extraction bioprocesses. Adv Biochem Eng Biotechnol 2003, 80:115-148. This review covers all of the integrated membrane assisted extractive processes. Stark D, von Stockar U: In situ product removal (ISPR) in whole cell biotechnology during the last twenty years. Adv Bioche Eng/Biotechnol 2003, 80:149-175. [This review covers all types of in situ product removal techniques.] Maas D, Gerigk MR, Kreutzer A, Weuster-Botz D, Wubbolts M, Takors R: Integrated L-phenylalanine separation in an E. Coli fed-batch process from laboratory to pilot scale. BioprocBiosynth Eng 2002, 25:85-96. Gerigk MR, Maas D, Kreutzer A, Sprenger G, Bonaerts J, Wubbolts M, Takors R: Enhanced pilot-scale fed-batch Lphenylalanine production with recombinant Escherichia coli by fully integrated reactive extraction. Bioprocess Biosyst Eng 2002, 25:43-52. Ru¨ ffer N, Heidersdorf U, Kretzers I, Sprenger GA, Raeven L, Takors R: Fully integrated L-phenylalanine separation and concentration using reactive-extraction with liquid centrifuges in a fed-batch process with E. coli. Bioprocess Biosyst Eng 2004, 26:239-248.

25. Thank you Prepared by, MISS RAHIMAH OTHMAN