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Microbial Biotechnology > Industrial Biotechnology > Flashcards

Industrial Biotechnology Flashcards

1
Q

At ScotBio, what organism do they use to produce blue food colouring

A

derived from Spirulina, a cyanobacteria

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2
Q

What is extracted from Spirulina and how?

A

the light-harvesting protein, phycocyanin is extracted and purified using photobioreactors

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3
Q

After purification, what happens to the protein

A

is formulated into powder & sold to distributers and manufacturers

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4
Q

how is cultivation of spirulina achieved

A

using open or closed systems, where the organism is grown to produce biomass or primary/secondary metabolites

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5
Q

Open system designs

A

raceway ponds
open ponds
tanks

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6
Q

advantages of open systems (2)

A
  • low cost
  • low energy requirement for culture mixing
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7
Q

disadvantages of open systems (3)

A
  • large areas must be scaled up for optimal growth (main cost associated with land)
  • contamination risks (weather conditions like wind may blow soil particles, chemicals e.g. pesticides, or algal grazers)
  • productivities are seasonal (in summer higher levels of sunlight, more growth)
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8
Q

Closed system designs

A

bioreactors & photobioreactors
- tubular reactors
- flat plate reactors
- air-lift reactors

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9
Q

Why are closed systems more suitable for culturing cyanobacteria (3)

A
  • allow for better control over environmental conditions required for growth (temp, light & nutrient level)
  • minimises risk of contamination (contained in sterile, sealed environment)
  • more efficient for space utilisation and resource consumption
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10
Q

At ScotBio, what do they use

A

Patented LED technology to provide optimal light levels required for Spirulina growth

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11
Q

disadvantages of closed systems (6)

A
  • bio-fouling (accumulation organic matter surfaces, affecting performance)
  • regular cleaning & maintenance necessary (issue if tank with corners, hard to CIP)
  • foaming (excess gas, foam reduces SA for gas exchange & light pene)
  • overheating due to lights or LEDs
  • growth of benthic algae (build up of DO, limit growth)
  • very high capital costs (design & installation - specialised equipment & monitoring)
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12
Q

Types of culture

A

batch
fed-batch
continuous

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13
Q

what is a batch culture

A

microorganisms are grown in a closed system with a fixed amount of nutrients, allowing them to consume this until depleted

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14
Q

what is a fed-batch culture

A
  • additional nutrients periodically added into the system
  • sustains growth over a longer time
  • allows for more control over nutrient level, optimising growth
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15
Q

what is a continuous culture

A

new nutrients are added while an equal amount of culture medium is removed (allows for constant growth of organisms, maintaining steady-state condition)

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16
Q

what are the main processes in downstream processing (7)

A

harvesting
lysis/extraction
separation
concentration
purification
formulation
drying

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17
Q

what is harvesting

A

the recovery of biomass from the culture, or removal of culture, leaving cells in reaction vessel

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18
Q

what is harvesting based on

A
  • filtration (using sieves, filter sheets or membrane filtration)
  • gravity-based (using separators like centrifuges or decanters)
  • sedimentation tanks
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19
Q

why is lysis required and what do we need to consider

A

to release intracellular target metabolites into the extractant
must consider thickness and composition of cell wall

20
Q

lysis methods

A
  • mechanical (agitation, high-pressure homogenisation)
  • chemical/biological (acid/alkali, enzymatic, solvent extraction)
  • physical/electromagnetic (free/thaw, ultrasound-assisted, microwave-assisted extraction)
21
Q

why must we consider the polarity/hydrophobicity of target compounds during lysis/extraction

A

if proteins are water soluble we must use an aqueous extraction buffer, so proteins can be solubilised and easier to extract

22
Q

why must we consider the stability of target compounds during lysis/extraction

A

if pigments are heat or pH-sensitive, proteins produced will be prone to denaturation if extraction process too vigorous

23
Q

what is separation for

A
  • to separate the complex mixture of molecules, cell debris and suspended solids left over from extraction
  • makes it easier to purify later
  • we may want to fractionate compounds into different product lines
24
Q

what are some gravity-based separation techniques (4)

A
  • separators (centrifuges & decanters)
  • sedimentation tanks
  • flocculants (aid removal of larger debris)
  • encourage precipitation of certain chemical groups
25
Q

how can distillation be used in the separation stage

A
  • can utilise differences in boiling points to separate liquids and volatiles from other liquids
  • freeze distillation can also be used to separate components (good for heat unstable compounds)
26
Q

different types of chromatography for separation (3)

A
  • reverse-phase
  • normal -phase
  • HILIC size-exclusive
27
Q

chromatography for separation

A
  • widely used technique to separate compounds
  • many different types & can be scaled
  • based on different compounds interacting with the stationary phase
  • or based on size
28
Q

why is concentration necessary

A

target compounds are often in very low concentrations, so we must make them amenable to processing, drying and shipping

29
Q

concentration techniques (3)

A

crossflow filtration
chromatography
evaporation

30
Q

what does the level of purification depend on

A
  • final application of product
  • legislation
31
Q

final applications for the product

A
  • technical grade - industrial use
  • laboratory grade - for use in teaching labs
  • food grade - for use in foods
  • reagent/BP/USP grade - for use in food, drug, body laboratory, & medical purposes
  • ACS grade - highest purity level, for use as analytical standards
32
Q

purification techniques

A
  • diafiltration
  • chromatography
  • sterile/microfiltration
33
Q

diafiltration

A
  • utilise ultrafiltration & nanofiltration membranes
  • removes salts & low MW molecules
  • repeatedly diluting & concentrating
  • increases the purity of retained molecules
34
Q

chromatography (for purification)

A
  • can bind (adsorb) compound to solid station phase
  • other compounds remain in mobile phase & pass through column
  • can then unbind analytes in another mobile phase
  • this concentrates and purifies them
35
Q

sterile/microfiltration

A
  • removes contaminating microbes and particulate matter
  • passes through filter with small pores
  • depth filtration (type of microfiltration) is commonly used to clarify liquids by removing certain contaminants
36
Q

purification

A
  • anti-solvent precipitation
  • salting out
  • isoelectric precipitation
37
Q

why do we often need to formulate the extract, concentrated & purified compound with excipients

A
  • improves functionality
  • improves bioavailability
  • increases stability & shelf-life
  • improves organoleptic qualities (smell, colour, taste)
38
Q

strategies for formulation

A
  • simply mixing excipients
  • homogenising mixtures to create micelles, emulsions or double emulsions
  • add gelling agents to create gels
  • formulate & dry compounds to micro-encapsulate them
39
Q

what is dried

A
  • whole biomass
  • cell extracts
  • purified compounds
40
Q

why are these compounds dried

A
  • increase shelf-life
  • aid micro encapsulation
  • help supply-chain logistics for transport
41
Q

what does drying choice depend on

A
  • heat stability of compounds
  • technoeconomics
  • other technical considerations
42
Q

methods for drying

A
  • drum drying
  • vacuum drying
  • spray drying
  • fluid bed drying
  • lyophilisation (freeze drying)
43
Q

drum drying

A
  • cheap
  • great for low value commodities
  • may damage heat sensitive compounds
44
Q

vacuum drying

A
  • economical
  • lower residence time than drum drying
45
Q

spray drying

A
  • allows fine control over final moisture content and particle size
  • very short exposure to high temperatures
46
Q

lyophilisation (freeze drying)

A
  • useful for very heat-sensitive compounds
  • expensive
  • slow
  • products need freezing beforehand

Microbial Biotechnology (10 decks)

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