Simulation Of BioprocessERT 315/4

5

TYPES OF BIOPROCESSAND

BIOPRODUCTS

Criteria to select appropriate Criteria to select appropriate biocatalystbiocatalyst

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?

Characteristics of biocatalystCharacteristics of biocatalyst

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

Bioreactor

Bioreactor

Bioreactor

Bioreactor

Bioreactor

Whole plant

Whole animal

Pure substrates

Simple media

Simple media

Complex media

Simple media

Fertilizer, CO2, various others

Various plant & animal materials

Certain parts of plants, animals and humans

Short

Short

Medium

Medium

Medium

Long

Long

Long

Simple

Medium

Medium

Medium

Medium

Complex

Complex

Complex

No

No

No

Yes

Possible

Possible

Yes

Possible

No

Small

Small

Medium

Small

Small

High

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

Biocatalyst Biocatalyst Enzyme BiotransformationEnzyme 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

-widely used in the production of fine chemicals and pharmaceuticals, e.g. vitamin C, amino acids, antibiotics, and steroids

-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

Metabolic Bioconversion using cell cultivationMetabolic Bioconversion using cell cultivation

- Classified into five groups: Bacteria, Fungi, Mammalian cells, insect cells, plant cells

(i) Bacteria-unicellar prokaryotes with a rigid cell wall-key determinants for their growth: media composition, temperature, 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) (80-105 °C)-optimum pH: 6.5-7.5

(ii) Fungi-divide into two subgroups: yeasts, molds-yeast: small, single cells that can grow as individual cells or clumps-used to produce alcohol in anaerobic fermentation, baker’s yeast, yeast extract as food additive-molds: multicellular, vegetative structure call mycelium, as usually highly-branched systems of tubules-mostly grown under aerobic conditions and the formation of a dense filamentous mycelium in the form of cell aggregates and pellets often cause oxygen-transfer problems-used for citric acid production, antibiotics, and riboflavin

(iii) Mammalian cells-produce correctly folded proteins and secrete them to the culture environment-carrying out required post-translational modifications of proteins, e.g. glycosylations-generally produce high-value proteins where are correct (native) three-dimensional structure is crucial-have complex nutritional requirements requiring serum, e.g. fetal calf serrum-grow quite slowly, with typical doubling times of 12-20 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 fir veterinary vaccines for the swine fever virus

(v) Plant cells-10 to 100 times larger than microbial cells and more sensitive to shear-slow metabolism, with doubling times of 20-100h resulting in low volumetric productivities even though high cells densities can be reached-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 PlantsTransgenic 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

Transgenic AnimalsTransgenic 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 TechnologiesExtractive 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

BioproductsBioproductsProduct Classifications/Characteristics-according to size, bioproducts can be divided into:a. Small moleculesb. Large moleculesc. Solid particles-Small moleculesi. sugars, amino acids, organic acids, vitaminsii. Molecular weight of 30-600 Da and a radius smaller than 1 nmIii divided into primary and secondary metabolites:a. Primary metabolites: sugar, organic alcohols, acids-produced in the primary growth face of the organismb. Secondary metabolites: formed at or near the beginning of the stationary phase, e.g. antibiotics and steroids-large moleculesi. proteins, nucleic acids, polysaccharidesii. Molecular weight of 103-106 Da and a radius typically larger than 1 nm-solid particlei. Whole cell like yeast, animal cell, ribosomes, virusesii. A radius of up to several µm-by the scale of production, bioproducts can be classified into:a. Bulk or commodity chemicals made at large scaleb. Fine chemicalsc. Pharmaceuticals made a smaller scale

-bulk chemicalsi. Produced in very large amounts (e,g, more than 1 000 000 tons per year)ii. Simple downstream processingiii. Sold at a relatively low priceiv. Medium purityv. Biocatalyst that grows in inexpensive media and reaches at a high productivity are necessary-pharmaceuticalsi. produced in a small amounts (as low as few kg per year)ii. high priceiii. used expensive media and complex equipment with low productivitiesiv. complex product separation and purification is acceptable for economic commercial production-fine chemicalsi. Used as intermediates and have application in a variety of industriesii. Annual production, price, and required purity lie between bucl chemicals and pharmaceuticals

Product classes-describe by its function (proteins, organic acids, lipid) or application (food and feed additives, pharmaceuticals, detergents, chemical intermediates, agriculturally used products)-organic alcohol and ketones- produced in anaerobic fermentations, from inexpensive carbon-energy sources such as glucose, starchy materials, molasses, sucrose containing materials-organic acids:-used as intermediates or as food additives-major organic acids produced are citric, lactic, gluconic acid.-amino acids:-the building blocks of protein and are connected via peptide bonds-used as food additives, feed additives, and in pharmaceuticals-most important amino acid are L-glutamic acid and L-lysine that are produced from molasses and starch hydrolysates-nucleic acids: -used as therapeutics, e.g. DNA vaccines, gene therapy-produced as short interference RNA molecules (sRNAi), aptamers (small DNA, RNA, or peptide molecules that bind with high specifity and affinity to DNA, RNA, or proteins-antibiotics:-frequent use in human and animal health-produced on fungal fermentation

-vitamins:-produced in bioprocesses, e.g. vitamin A, C, E, and the B vitamins-biodegradable biopolymers:-plastics derived from renewable material-common biopolymer are polyhydroxyalkanoates (PHA) and polyhydroxybutyrate (PHB)-dextran and xanthan:-industrially produced microbial polysaccharides-used as thickening, gelatinizing, suspending agents in food and pharmaceuticals-carotenoids:-natural pigments (yellow or red color)-produced by microorganisms-pesticides-lipids:-including fats, oils, waxes, phospholipids, steroids-commercially produced lipid: prostaglandins, leukotrienes, thromboxane-proteins:-characterized by four level of structure: primary structure (linear amino acid sequence), secondary, hydrogen-bonded structure (alpha helix and beta sheet), tertiary (folding pattern of hydrogen-bonded and disulfide-bonded structures), quaternary structure (formation of homo- and hetero- multimeric complexes by individual protein molecules)-two major applications: as industrial enzymes, as therapeutic and diagnostic proteins

Raw materialsRaw 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 mineralssuch as magnesium and potassium ions-carbon-energy source:-dominant requirement as it 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, amino acid, urea, complex materials-oxygen and hydrogen:-20% of the cell mass (oxygen), 8% (hydrogen)-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 ( almost always needed), copper, sodium, calcium, boron(needed only under specific growth conditions)-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-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.

Waste treatment, Reduction and Waste treatment, Reduction and RecyclingRecycling

Eco

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ic

savi

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Eco

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cost

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1. Avoid waste formation

2. Reduce waste formation

3. Extend material use

4. Recycle material

5. Downcycle material

6. Treat waste/energetic recovery

7. Safe water disposal

Steps of waste avoidance and treatment

1. 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 as much as useful

2. Reduce waste formation-reuse of material, e.g. a chromatography resin can be used for multiple cycles, the annual amount of waste is significantly reduced

3. Recycle material use-recycling of an organic solvent used in an extraction step: good of cost-effective recycling-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 (downcycling)

4. Treat waste/energetic recovery-have to be treated or disposed safely-some energy is produced during the treatment (e.g. incineration)

Types of wasteTypes of waste-gaseous stream:-cause from the exhaust air from a bioreactor (contains air, CO2, and water), distillation and evaporation steps (e.g. associated with crystallization)

-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-can be used as animal feed or organic fertilizer or treatment plant

-liquid waste:-treated in a biological sewage treatment plant at the production site of the bioprocesses-also can be released to the municipal sewer system-pretreatment is necessary – at a high low or low pH, the liquid has to be neutralized byadding base or acid

Overall development processOverall development processD

evel

op

men

t st

eps

Development processProduct idea

Literature/patent review

Biocatalyst screening

Biocatalyst optimization

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

• Goal of every process development: product• Product- should have a market, potential market, sufficient size that economically justifies

the required environment• Required literature and patent review – to clarify if there are similar products already on

the market or in the development• Find a biocatalyst (organism or an enzyme that catalyses the formation of the desired

product)- to reach an economically feasible product yield and concentration• The medium and reaction conditions are adjusted enable the best performance of the

catalyst- medium should be as simple and inexpensive as possible and the reaction conditions should provide the best environment for the biocatalyst

• The process flow diagram (PFD) put together all the operation• All unit operations have to be chosen and connected in an efficient and robust manner- to

maximize the overall yield• Determined the size plant- validate the market share of the product estimated at the

beginning• Scale up the process• Plan and implement the clinical trials for pharmaceuticals• Build a process models to estimate the material balance, energy consumption, labor

requirement and equipment needed

Upstream processing

BioreactionDonwstream processingRaw

material

Process

Waste Treatment/

disposal

ConsumablesUtilities Labor

Finalproduct

6

SustainabilityAssessment

Sustainability AssessmentSustainability AssessmentWhat 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

Economic Assessment

Bioengineering

Conversion, yield Process flow diagram

Volume/massof product

Raw materials

Utilities/waste

Labor

Consumables

Operating cost

Purchase equipmentcost

Capital investment

Equipment prices

Multipliers

1.Estimate the capital investment2.Operating costs can be derived from the different cost items

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:i. Equipment purchase costii. Estimation of total capital investment:a. Direct cost- the purchased equipment that need to be installedb. Indirect cost- a number of planning costs, like the preparation of design books that document the process, the design of equipment, etciii. Multiplier values –for a realistic estimation of the capital investment, derived from the empirical data and are different for different process typesiv. 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

v. 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

Operating-cost simulationOperating-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:1. Raw materials – the list of raw materials and the amounts consumed 2. Consumables- all material and equipment parts that have to be replaced from time to time3. Labor- determined by the operator hours and the hourly wage4. Operating supplies- including clothing, tools, and protective devices for the workers and also everyday items needed to run the plant5. Laboratory, quality control, and quality assurance6. Utilities- energy consumption for heating, cooling, evaporation/distillation, aeration, etc7. Waste treatment and disposal8. Royalty expenses- single unit operations or even the whole process that has to be covered by a patent owned by others-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 money2. Maintenance and repair3. 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-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

Profitability AssessmentProfitability 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

processi. 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

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 100v. 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/ROITime value of money: time-value of the earned money NPV=ΣAj/(1+i)j

n

j+1

Environmental Assessment-to identify the environmental ‘hot spots’ of the process

Process characteristicsModeling and simulation

Component propertiesImpact categories

Material balance ABC classification

Mass indices (MI)Environmental factors

(EF)

Environmental indices(EI)

Impact categories Process Components

Assessment structure of the method

Structure of the method-two starting point:a. 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 outputcomponents (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 concerningenvironmental, 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

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

Environmental Factor component i,

EFi (index points/kg)

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

Raw Material Availability

Land Use

Complexity of Synthesis

Thermal Risks

Acute Toxicity

Chronic Toxicity

Ecotoxicity

Global Warming Potential

Ozone Depletion Potential

Acidification Potential

Photochemical Ozone Creation Potential

Odor

Eutrohication Potential

Organic Carbon Pollution Potential

Resources

Grey Input

Component Risk

Organism

Air

Water/Soil

Environmental FactorInput component

Environmental FactorOutput component

Impact categories Impact groups Environmental Factors

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

I

I

I

I/O

I/O

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 30-100 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: 100-1000mg/m3; IDLH: 100-1000 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

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

I/O

I/O

O

O

O

O

O

O

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

POCP<@ 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

Social AssessmentSocial AssessmentAspect 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

-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

-apprenticeship

-voluntary training offerings

-identification of training needs

-incorporation of employee expectations

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

-Used information system

-Control of knowledge exchange

-Employee involvement in decision-making

-degree of innovation

-product readiness and marketability

-estimated market penetration

-number and types of patents

-product acceptance

-usage of genetic engineering methods

-social standards in supply chain

-societal benefits

-used communication channels

-reporting of core activities to neighbors

-targeted dialogue partners

-measures taken to promote dialogue