Presentation is loading. Please wait.

Presentation is loading. Please wait.

Simulation Of Bioprocess ERT 315/4

Similar presentations

Presentation on theme: "Simulation Of Bioprocess ERT 315/4"— Presentation transcript:

1 Simulation Of Bioprocess ERT 315/4


3 Criteria to select appropriate biocatalyst
What yield, product concentration, and productivity can be reached? What substrate can be utilized, what additional media components are required, and how does it all effect downstream processing? What by-products are formed and how do they affect yield and downstream processing? What are the challenges in biocatalyst preparation, storage, propagation, security, and safety? What are the optimal reaction conditions, e.g. temperature, oxygen supply, shear sensitivity, foam formation, etc? How well do we understand the reaction mechanisms, are they robust and genetically stable? If the product is expressed intracellularly, how is it extracted? How do we purify the desired product form the many impurities in the process?

4 Characteristics of biocatalyst
Production device Raw material Time-scale Purification Complex protein structure Viral/prion risk Process examples Enzymes Bacteria and yeast Fungi Mammalian cells Plant cells Transgenic plants Transgenic animals Extractive technology Bioreactor Whole plant Whole animal Pure substrates Simple media Complex media Fertilizer, CO2, various others Various plant & animal materials Certain parts of plants, animals and humans Short Medium Long Simple Complex No Yes Possible Small High Cyclodextrin, Acrlyamide, L-dopa Lysine,Vitamin B2, Insulin Citric acid, antibiotics Monoclonal Antibodies, Interferons Taxol, shikonin, Methyldigoxin Antibodies, antibody Fragments, HAS, PHB Α1-antirypsin, HAS, Lactoferrin Plasma,components,taxol

5 Biocatalyst Enzyme Biotransformation
-enzymes: protein with a unique three-dimensional structure able to bind a substrate, usually but not always a small molecule, and catalyze a specific reaction, similar to chemical catalysis but under mild conditions of temperature and pressure -classified in six groups according to the chemical reaction: oxido-reductase, transferases, hydrolases, lyases, isomerases, ligases -highly selective and specific in the reaction: regio-, stereo- and enantioselective -can be in solution or immobilized -five major of reactions where enzymes are used industrially: hydrolysis of proteins, palysaccharides,esters,amides, nitriles, epoxides; synthesis of esters, amides, glycosides; carbon-carbon bond formation; reduction reaction; oxidation reactions

6 Metabolic Bioconversion using cell cultivation
- Classified into five groups: Bacteria, Fungi, Mammalian cells, insect cells, plant cells Bacteria -unicellar prokaryotes with a rigid cell wall -key determinants for their growth: media composition, temp., gaseous environment, pH -show a range responses to oxygen: aerobic bacteria- require oxygen for their growth; anaerobic bacteria- grow only at the absence of oxygen; facultatively-anaerobic bacteria- able to grow under both conditions -temperature optimal for growth: psychrophiles (20-30 °C), mesophiles (30-40 °C), thermophiles (45-60 °C), extreme thermophiles (extremophiles) ( °C) -optimum pH:

7 (ii) Fungi -divide into two subgroups: yeasts, molds -yeast: small, single cells that can grow as individual cells or clumps -molds: multicellular, vegetative structure call mycelium, as usually highly-branched systems of tubules (iii) Mammalian cells -produce correctly folded proteins and secrete them to the culture environment -grow quite slowly, with typical doubling times of h -temperature : 37 °C, pH: 7.3 -mammalian cell product: monoclonal antibodies, interferons, vaccines, erythropoietin (iv) Insect cells -produce recombinant proteins less expensively and more quickly than mammalian cells and at high expression levels -typically grow at around 28 °C, and pH 6.2 -used for veterinary vaccines for the swine fever virus

8 (v) Plant cells -10 to 100 times larger than microbial cells and more sensitive to shear -slow metabolism, with doubling times of h -cultivate as a callus or a lump of undifferentiated plant tissue growing on a solid nutrient medium or as aggregated plant cells in suspension -used to produce secondary metabolites, anticancer drug paclitexel (taxol), recombinant proteins of high value Transgenic Plants -genetically modified plants to produce a wide variety of products -the expression can take place in the whole plant or only in a certain part as in the seeds -commonly used plants: tobacco, potato, rice, wheat -inexpensive, easy to scale-up, free of human pathogens

9 Transgenic Animals -reduce the dependency on the seasonal and geographical conditions, post-translational modifications are more likely to mimic the native structure -usually done by injecting exogenous DNA into egg cells to produce a vital embryo that is later able to express the desired product Extractive Technologies -comprise all processes where a product is extracted from natural material -Used in the extraction of pharmaceuticals from human or animal blood and from plant material -the products usually chemically complex non-protein materials

10 Bioproducts Product Classifications/Characteristics
-according to size, bioproducts can be divided into: Small molecules, Large molecules and Solid particles -Small molecules sugars, amino acids, organic acids, vitamins Molecular weight of Da and a radius smaller than 1 nm Iii divided into primary and secondary metabolites: Primary metabolites: sugar, organic alcohols, acids-produced in the primary growth face of the organism Secondary metabolites: formed at or near the beginning of the stationary phase, e.g. antibiotics and steroids -large molecules proteins, nucleic acids, polysaccharides Molecular weight of Da -solid particle Whole cell like yeast, animal cell, ribosomes, viruses A radius of up to several µm

11 Produced in very large amounts Simple downstream processing
-by the scale of production, bioproducts can be classified into: Bulk or commodity chemicals made at large scale, Fine chemicals and Pharmaceuticals made a smaller scale -bulk chemicals Produced in very large amounts Simple downstream processing Sold at a relatively low price -pharmaceuticals produced in a small amounts high price used expensive media and complex equipment with low productivities -fine chemicals Used as intermediates and have application in a variety of industries Annual production, price, and required purity lie between bulk chemicals and pharmaceuticals

12 Product classes -describe by its function (proteins, organic acids, lipid) or application (food and feed additives, pharmaceuticals, detergents, chemical intermediates, agriculturally used products) i. organic alcohol and ketones produced in anaerobic fermentations ii. organic acids: -used as intermediates or as food additives -major organic acids produced are citric, lactic, gluconic acid. iii. amino acids: -the building blocks of protein and are connected via peptide bonds -used as food additives, feed additives, and in pharmaceuticals iv. nucleic acids: -used as therapeutics, e.g. DNA vaccines, gene therapy v. antibiotics: -frequent use in human and animal health -produced on fungal fermentation

13 vi. vitamins: -produced in bioprocesses, e.g. vitamin A, C, E, and the B vitamins vii. biodegradable biopolymers: -plastics derived from renewable material -common biopolymer are polyhydroxyalkanoates (PHA) viii. dextran and xanthan: -industrially produced microbial polysaccharides -used as thickening, gelatinizing, suspending agents ix. carotenoids: -natural pigments (yellow or red color) -produced by microorganisms x. pesticides xi. lipids: -including fats, oils, waxes, phospholipids, steroids -commercially produced lipid: prostaglandins, leukotrienes, xii. proteins

14 Raw materials -Water: -dominant raw material
-other component of the reaction medium: macronutrients and micronutrients -macronutrient: -needed in concentrations larger than 10-4 M -including carbon–energy source, oxygen, nitrogen, phosphate, sulfur, and some minerals such as magnesium and potassium ions -carbon-energy source: -provides the carbon for biosynthesis as well as energy derived by its oxidation -Typically used carbons sources: glucose, starch, corn syrup, molasses. soybean oil, palm oil, ethanol, methanol -50% is incorporated in the biomass, and remaining 50% is used to derived energy for biosynthesis -nitrogen: -accounts for 10-14% of the dry cell mass -most widely used are ammonia and ammonia salts, proteins,

15 -oxygen and hydrogen:-20% of the cell mass (O2), 8% (H2)
-phosphorus: 3% of cell dry weight and is provided by phosphate salts -sulfate: 0.5% of cell mass is added as sulfate salts or with amino acids contained in complex media -magnesium and potassium ions: provided as inorganic potassium and magnesium sulfate -micronutrients: -required in low concentrations: -including iron, zinc, manganese,copper, sodium, calcium, boron -added as inorganic salts -also can be classified into defined or synthetic media and complex or natural media -defined media: -contain specific amounts of pure chemicals with a known composition -complex media: -include one or more natural materials whose chemical composition is not exactly known and which may vary with source of time

16 -natural media: -cheaper -cause less reproducible fermentation and more complex downstram processing -bacteria and fungi: need only a relative simple media and very low cost -mammalian cells: more complex medium is necessary, need serum as required ingredient (complex media) or not (synthetic media) -plant cell: require a carbohydrate cell source, inorganic macronutrients and micronutrients.

17 Waste treatment, Reduction and Recycling
Avoid waste formation Reduce waste formation Extend material use Recycle material Downcycle material Treat waste/energetic recovery Safe water disposal Ecological costs Economic savings Steps of waste avoidance and treatment

18 Avoid waste formation -to avoid the formation of waste -if feasible and cost-effective, subsequent treatment is unnecessary -if cannot be prevented completely, try to reduce it 2. Reduce waste formation -reuse of material 3. Recycle material use -recycling of an organic solvent used in an extraction step -Compared the amount recycle and the amount of material and energy necessary for the recycling to decide it is economically and environmentally favorable -if cannot be recycled because the purification becomes too expensive, used another purpose that requires less purity 4. Treat waste/energetic recovery -have to be treated or disposed safely

19 Types of waste -gaseous stream: -cause from the exhaust air from a bioreactor (contains air, CO2, and water), distillation and evaporation steps -solid waste: -categorized as hazardous and non-hazardous waste -hazardous waste: -containing heavy metals or highly toxic substances -need special treatment or disposal with high-safety manners- cause higher costs -non-hazardous waste: -wet biomass -if a recombinant organism is used, sterilization of the material is necessary, usually by heat -liquid waste: -treated in a biological sewage treatment plant at the production site of the bioprocesses

20 Overall development process
Literature/patent review Biocatalyst screening Biocatalyst optimization Development steps Medium and reaction condition optimization Selection of downstream steps Identification of PFD Optimization of unit operations Plant size Scale-up : Lab – technical - industrial Approval, clinical trials Process modeling and uncertainty analysis Economic and environmental assessment Development process Product idea

21 Process Upstream processing Bioreaction Donwstream processing Raw material Final product Consumables Utilities Labor Waste Treatment/ disposal

22 Sustainability Assessment

23 Sustainability Assessment
What is “sustainability”? -the development that meets the needs of the present without compromising the ability of the future generations to meet their own needs (Brundtland, 1980s) -the optimal growth path that maintains economic development while protecting the environment and optimizing the social conditions with the boundary of relying on limited, exhaustible natural resources -a business approach that creates a long-term shareholder value by embracing opportunities and managing risks deriving from economic, environmental, and social development Sustainability Economic Ecological Social

24 Economic Assessment 1.Estimate the capital investment
Bioengineering Conversion, yield Process flow diagram Equipment prices Raw materials Volume/mass of product Utilities/waste Purchase equipment cost Labor Consumables Multipliers Operating cost Capital investment 1.Estimate the capital investment 2.Operating costs can be derived from the different cost items

25 Capital cost estimation
-total amount of money that has to be spent to supply the necessary plant (the fixed capital investment) plus the working capital that is needed for the operation of the facility -how to estimate: Equipment purchase cost Estimation of total capital investment: Direct cost- the purchased equipment that need to be installed Indirect cost- a number of planning costs, like the preparation of design books that document the process, the design of equipment, etc Multiplier values –for a realistic estimation of the capital investment, derived from the empirical data and are different for different process types Prices indices- changes of equipment price over time due to inflation/deflation or market conditions Present prices = Price at t0 x index value today/ index value at t0 Scale-up Factors- the cost of a single piece of equipment or a complete plant changes when its capacity is changed C2= C1 x (q2/q1)0.6, where C1: coast of a plant with a certain capacity q1

26 Operating-cost simulation
-the total of all costs operating the plant and recovering the capital investment, i.e. the annual amount of money necessary to produce the product and pay back investment cost -Divided into variable, fixed, plant overhead cost -Variable cost: Raw materials – the list of raw materials and the amounts consumed Consumables- all material and equipment parts that have to be replaced from time to time Labor- determined by the operator hours and the hourly wage Operating supplies- including clothing, tools, and protective devices for the workers and also everyday items needed to run the plant Laboratory, quality control, and quality assurance Utilities- energy consumption for heating, cooling, evaporation/distillation, aeration, etc Waste treatment and disposal Royalty expenses- single unit operations or even the whole process that has to be covered by a patent owned by others

27 -Fixed Cost (Facility-dependent cost)
1. depreciation- a capital investment that was need to built a plant and this investment has to be paid by charging an annual amount of money 2. Maintenance and repair 3. Insurance and local taxes-derived from the direct fixed capital (DFC) 4. Rent and interests -Plant overhead cost –factory expenses, or plant overhead costs caused by the operation of facilities that are not directly related to the process, e.g. medical service, safe and protection Others: -general expenses- the general expenses that has to cover to manage the company, to sell product and computer support. Including administration, distribution and marketing, research and development -Unit production cost (UPC)- the total product cost allocated to the annual amount of product

28 Profitability Assessment
-revenues: sum of all sales of the main and side product of a process within a certain time period usually a year. For a single-product facility, the revenue r for year j is : rj= mjxpj , where mj is the amount of product sold in year j and pj the (average) price realized in this year -measurement of profitability- a number of indices that are used to evaluate the profitability of a process i. The gross profit in year j (Gj) is the annual revenue rj minus the annual total product cost including depreciation: Gj=rj-cj ii. The net profit in year j (Nj) is the gross profit minus the income tax. The income tax is determined by the tax rate Ф Nj = (rj-cj)x(1- Ф) = Gjx(1- Ф) iii. The net cash flow in year j (Aj) is the sum of net profit and the depreciation dj of the year Aj=Nj+dj n j+1

29 iv. The return on investment (ROI) is the ratio of profit to investment and measures how effectively the company uses its invented capital to generate profit. Usually calculated using the net profit and the total investment (TCI) and is shown in percentage value: ROI=Nj/TCI x 100 v. The payback period (PBP) is the length of time necessary to pay out the capital investment by using the annual cash flow that return to the company’s capital reservoir. In most cases, the direct fixed capital (DFC) is used for this index PBP=DFC/Aj The PBP also can be calculated using the TCI and the net profit PBP=TCI/Nj=100/ROI Time value of money: time-value of the earned money NPV=ΣAj/(1+i)j

30 Environmental Assessment
-to identify the environmental ‘hot spots’ of the process Process characteristics Modeling and simulation Component properties Impact categories Material balance ABC classification Mass indices (MI) Environmental factors (EF) Environmental indices (EI) Impact categories Process Components Assessment structure of the method

31 Structure of the method
-two starting point: Process and characteristics by Superpro Designer model -the result is the material balance of the process -from the material balance, Mass Index (MI) can be calculated for all input (states how much of component is consumed to produce a unit amount of the final product) and output components (how much of a component is formed per unit final product) b. From the component properties -classified into A, B and C classes that represent environmental relevance (high, medium, low relevance) from the 15 impact categories (IC) that may have a negative effect on human health and the environment -the IC are allocated to six impact groups, representing an important field Concerning environmental, health or safety aspects -numerical values for the classes A, B and C, and a weighting factor (=environmental factor) is derived from its classifications in the impact groups -next step: -Link the amount of the components in the mass balance with Environmental Factors -The resulting Environmental Index (EI) helps to identify those components that are environmentally most relevant in the process

32 Weighting factors/indices Calculation
Mass Index component i, MIi (kg/kg P) mi= amount of component i (kg); mp= amount of final product (kg P) Mass Index process, MIprocess (kg/kg P) Environmental Factor component i, EFi (index points/kg) Via arithmetic average; as EFMv,i,In/EFMV,i,Out IGj,i= value of component i in Impact Group j; j= Number of impact Groups Via multiplicatiom; as EFMult,i,In and EFMult,i,Out Environmental Index component i, EIi (index points/kg P) (as EIi,In or EIi,out) Enviromental Index process EIprocess (index points/kg P) (as EIIn or EIOut) Generak Effect Index Process GEI (nondimensional) MI=mi/mp MIprocess,In =Σmi/mp MIprocess,Out=1+Σmi/mp EFMv,i=(IG1,i+IG2,i+IG3,i+IG4,i)/j EFMult,i=πIGj,i EIi=EFi,x mi/mp=EFi x MIi EIprocess=ΣEIi GEI=EIprocess/MIprocess

33 Environmental Factors
Impact groups Impact categories Environmental Factors Raw Material Availability Resources Land Use Environmental Factor Input component Complexity of Synthesis Grey Input Thermal Risks Component Risk Acute Toxicity Chronic Toxicity Organism Ecotoxicity Global Warming Potential Ozone Depletion Potential Acidification Potential Air Photochemical Ozone Creation Potential Environmental Factor Output component Odor Eutrohication Potential Water/Soil Organic Carbon Pollution Potential

34 Impact category I/O Class A Class B Class C Raw Material Availability Land use Critical material used Complexity of the Synthesis Thermal risk Acute toxicity I Only fossil, Predicted exhaustion within 30 years >100m2/kg Critical materials like heavy metals, AOX, PCB used or produced in stoichiometric amounts >10stages R 1-4, 9, 12, 15-17, 44; EU; F+, E; NFPA F+R: 3,4 EU:T+; R 26-28,32; CH-poison class: 1,2; NFPA H:4; WGK 3; ERPG:<100mg/m3; IDLH: <100mg/m3 Only fossil, predicted exhaustion in years >10m2/kg and<100m2/kg Critilcal materials involved in sub-stochimetric amounts 3-10stages R 5-8, 10, 11,14,18,19, 30; EU: F, O; NFPA F+R:2 EU: T, Xn,Xi,C; R 20-25, 29, 31,34-39,41-43,65,66,67; NFPA H: 2,3; WGK 2; ERPG: mg/m3; IDLH: mg/m3 Exclusively renewable, or guaranteed long term supply (>100 years) <10m2/kg No critical compounds involved <3 stages NFPA F+R: 0,1 CH-poison class: 5; NFPA H: 0,1; WGK 1, ERPG: >1000mg/m3; IDLH:>1000mg/m3

35 Impact category I/O Class A Class B Class C Chronic Toxicity Ecotoxicity Global Warning Potential Ozone depletion potential Acidification potential Photochemical ozone creation potential Odor Euthrophication potential Organic carbon pollution potential O MAK:1mg/m3; IARC;1,2A;R 45-49,60,61 EU:N;R 50 WGK 3; GWP>20 ODP>0.5 AP>0.5 POCP>30 or NOx N-content>0.2 or P-content>0.05 MAK:1-10mg/m3;IARC;2B,3; R 33,40,62,63; EU: T,T+,Xn; CH-poison class: 1,2 R 51-58; WGK 2 GWP<20 ODP<0.5 AP<0.5 30>POCP>2 Odor threshold <300mg/m3 N-content<0.2 and P-content<0.05 ThOD>0.2 g O2/g substrate MAK:>10mg/m#;IARC: 4: CH-poison class: 3,4,5 WGK ! Or no water hazard No global warming potential No ozone depletion potential No acidification potential or no effect known Odor threshold >300mg/m3 or no odor compound without N and P Compound without N and P ThOD<0.2g O2/g substrate or no organic compound

36 Social Assessment Aspect Social Indicators Technology development
Technology application Health and safety Quality of working conditions Employment Education and training -risk group of biological substances -risk factors for health and safety -voluntary health measures -quality of health and safety management -working time arrangement -degree of psychological strain -Percentage of women in leading positions -measures taken to improve working conditions -safeguarding of jobs -continuity of job creation effects -regions of job creation -extent of job creation -focus on employee training -quality of human resource management -identification of training needs -incorporation of employee expectations -job security levels -amount of hazardous substances -voluntary health measures during application -voluntary health measures during usage -working time arrangements -percentage of women in leading positions -apprenticeship -voluntary training offerings

37 Aspect Social Indicators Technology development Technology application Knowledge management Innovation potential Product acceptance and social benefit Societal dialogue -Degree of knowledge exchange -Used information system -Control of knowledge exchange -Employee involvement in decision-making -commercial exploitation potential -contribution to scientific debate -management of patents and licenses -number and types of patents -stakeholder involvement -usage of genetic engineering methods -social standards in supply chain -societal benefits -voluntary provision of information -reporting of core activities to neighborhood -stakeholder involvement in strategic decision making -communication channels to political debates -aspects of knowledge exchange -degree of innovation -product readiness and marketability -estimated market penetration -product acceptance -used communication channels -reporting of core activities to neighbors -targeted dialogue partners -measures taken to promote dialogue

Download ppt "Simulation Of Bioprocess ERT 315/4"

Similar presentations

Ads by Google