KC32003 BIOPROCESS PRINCIPLES

LECTURE 1: INTRODUCTION - Streak plate method

 Bioprocess engineer

- Study of biological and biochemical principles

 Bioprocess products: animal feed, cosmetic, fuel,
- Pour plate (loop dilution) method
pharmaceutical, chemicals, polymers, paper, food

LECTURE 2: BIOPROCESS INSTRUMENTATION/

LABORATORY TOOLS
1. Procedures for investigating and characterizing
microorganisms
 Inoculation: introduction of a sample into a container
of media to produce a culture of observable growth
- Placing a sample into a container of medium (S/L)
that supplies nutrients for growth
- Purpose: to increase visibility
- Pour plate and spread plate methods: - Spread plate method

 Inspection: observation; macroscopic (growth: colour,

texture, size) and microscopic (cell: shape, size,
motility)
 Identification: macroscopic and microscopic
appearance, biochemical tests, genetic characteristics,
immunological testing
 Disposal of Cultures
- Steam sterilization
- Incineration

2. Media: Providing nutrients in the laboratory

 Physical state
- Liquid
- Semisolid
- Solid (Can or cannot be liquefied)
 Incubation: exposing inoculated medium to optimal
 Chemical composition
growth conditions, usually a few hours or day
- Synthetic (chemically defined): contains pure organic/
- To promote multiplication and produce the actual
inorganic compounds in an exact chemical formula
culture
- Non-synthetic (complex, not chemically defined):
 Isolation: separating one species of bacteria from
contains at least one ingredient that is not chemically
another definable
- An individual bacterial cell will grow into a colony, if it
 Functional type
has space on a nutrient surface
- General purpose: grows a broad range of microbes
- Pure culture: single known species of microorganism
- Enriched: contains complex organic substances (e.g.:
- Mixed cultures: two or more identified species of
blood, serum) required by fastidious microbes
microorganisms
- Selective: inhibits growth of some microbes and
- Contaminated culture: once pure/mixed culture that
encourage growth desired microbes
has unwanted microbes growing
- Differential: allows growth of several types of
microbes, displays visible differences among them
 - Selective and differential media:
 Miscellaneous media
- Reducing medium: contains a substance that
absorbs oxygen/slows penetration of oxygen into
medium; used for growing anaerobic bacteria
- Carbohydrate fermentation medium: contains sugars
that can be fermented, converted to acids, and a pH
indicator to show this reaction
3. Microscope
 Magnification: ability to enlarge objects
- Objective lens forms the magnified real image
- Real image is projected to the ocular where it is
magnified again to form the virtual image
 Resolution: ability to show details
- Minimum distance at which two distinct points of a
specimen still can be seen
 Optical microscope
- Bright-field: specimen is darker than surrounding field
- Dark-field: brightly illuminated specimens surrounded
by dark fields
- Phase-contrast: transforms subtle changes in light
waves passing through the specimen into differences
in light intensity
 Fluorescence microscope
- Has a UV radiation source and filter that protects the
viewer’s eyes
- Uses dyes that emit visible light when bombarded
with shorter UV rays
 Electron microscopy
- Transmission electron microscope: transmits
electrons through the specimen (darker: thicker and
denser parts; lighter: transparent, less dense parts)
- Scanning electron microscope: provides detailed 3D
view
 Specimen preparation for optical microscopes
- Condition of specimen
- Aims of examiner
- Type of microscopy available
LECTURE 3: CELL STRUCTURE AND
CLASSIFICATION OF MICROBES
 Eukaryotic cell structure (external and boundary)
- Appendages: flagella and cilia (motility)
- Glycocalyx: outermost boundary that comes into
direct contact with environment (adherence,
protection, signal reception)
- Cell wall: provides structural support and shape
- Cell membrane: permeability barrier in transport
 Eukaryotic cell structure (internal)
- Nucleus: bounded with a double membrane (stores
DNA, coordinate cell’s activities)
- Endoplasmic reticulum: RER: protein synthesis,
modification of protein structure; SER: make cellular
products, regulates and releases calcium ions and
processes toxins
- Golgi apparatus: made up of cisternae (transporting,
modifying, and packaging proteins/lipids into vesicles
for delivery)
- Mitochondria: powerhouse of the cell
- Ribosomes: synthesise protein molecules
- Cytoskeleton: gives shape, structural support
 Prokaryotic cell structure (external and boundary)
- Appendages: flagella and pili (motility), fimbriae
(adherence), sex pili (share gene with other bacteria)
- Glycocalyx: thick outer covering of plasma membrane
(protects plasma membrane and cell, adherence)
- Cell wall: structural support, protection, filtering
mechanism, pressure vessel
- Cell membrane: protection, maintains cell shape,
regulate and control flow of materials in and out of cell
 Prokaryotic cell structure (internal)
- Cytoplasm: control what goes in and out of cell
- Ribosomes: smaller than eukaryotic ribosomes (build
proteins)
- Inclusion body: non-membrane bounded structure
located in cytoplasm (acts as a storage vessel)
- Nucleoid/chromosomes: single large circular strand
of DNA
- Plasmid: circular rings of DNA (carry auxiliary
information and genes for antibiotic resistance)
- Endospore: resistant to UV and gamma radiation,
desiccation, lysozyme, temperature, starvation, and
chemical disinfectants (ensure survival of bacterium)
 Taxonomy
- Organizing, classifying, and naming living things
- Classification, nomenclature, identification
 Usage of microorganisms
- Biotechnology: production of food, drugs, and
vaccines using living organisms
- Genetic engineering: manipulation of organism genes
to make new products
- Bioremediation: using living organisms to remedy an
environmental problem
 Microbial roles in infectious diseases
- Parasites: lives in or on a host and damages the host
- Pathogen: an organism that causes diseases
 Bacteria
- All bacteria are prokaryote, smallest microorganism
- Shape
Cocci Bacilli Spirilla Vibrio
- Cell wall is made up of peptidoglycan
- No well-developed nucleus, nuclear material is
scattered in the cytoplasm
- May have flagella to help in movement
- Nutritional: water, carbon and nitrogen sources,
inorganic salts
- Habitat range: psychrophilic (20 ⁰C), mesophilic (25 –
40 ⁰C), thermophilic (55 – 80 ⁰C)
- Reproduction/growth:
- Reproduces by binary fission
- Importance: producers and decomposer, enzyme for
medicine, F&B productions, nitrogen fixation
 Fungi
- Eukaryotic, heterotrophs, acquire food by absorbing
dissolved molecules, principal decomposers
- Nutritional: saprophytes (obtain food from dead
organic materials), parasites (obtain food from host),
symbionts (obtain organic food from partner and in
turn absorb inorganic nutrients and water)
- Reproduction: sexually/asexually, produces spores
- Importance: medicine (antibiotic), food (yeast),
agriculture (increasing soil fertility)
- Pathogenic: aspergillus (allergic), Cryptococcus (c.
neoformans, c. meningitis)
 Protozoa
- Eukaryote, nucleus is enclosed in a membrane
- Pellicle (shape), cytosome (ingesting F/S particles),
subpellicular tubules (motility)
- Nutritional: holozoic (require organic materials)
- Reproduction:
- Life cycle:
- Importance: predators of algae and bacteria,
producers in water ecosystem, decomposer
- Pathogenic: brain infection, intestinal, cardiovascular
 Helminths (parasitic worms)
- Relatively large, have well-developed organ
- Flatworms (flattened body covered with plasma
membrane) and roundworm (cylindrical body covered
with cuticle)
- Nutritional: parasitic (obtain nutrients from host)
- Habitat range: inside animal or plant
- Reproduction: either hermaphrodite or have
differentiated sex (produce eggs)
- Life cycle: 1 to 8 years
- Importance: treat overactive immune response
- Pathogenic: intestinal problem, elephantiasis,
 Algae
- Mostly eukaryotes, photosynthetic
- Nutritional: photoautotrophic (euglenoids),
heterotrophic (fire algae), holozoic (golden-brown
algae)
- Habitat range: aquatic environment
- Reproduction:
 - Importance: high biofuel yields, purify wastewater,  Nutritional types
biomass source, consumes carbon dioxide - Heterotrophs: obtain carbon in organic form
- Pathogenic: harmful to aquatic/land animals, blocking - Autotrophs: obtain carbon from carbon dioxide
of photosynthesis, contamination of water supply - Chemotrophs: gain energy from chemical
 Cell construction compounds
- Carbohydrates: supply energy to all cells in the body - Phototrophs: gain energy through photosynthesis
- Proteins: structure, function, and regulation of the - Photoautotrophs: gain energy from organic material
body’s tissues and organs synthesised through photosynthesis
- Lipids: storing energy, signalling, structural - Chemoautotrophs: synthesise organic food from
component of cell membrane inorganic compounds
- Nucleic acid: make up genetic information in living - Saprobe: gain nutrients from non-living or decaying
things organic matter
- Parasites: gain nutrients from host at its expense
LECTURE 4: MICROBIAL NUTRITION AND - Photoheterotrophs: use light for energy, but cannot
METABOLISM use carbon dioxide as sole carbon source
1. Microbial nutrition  Transport: movement of chemicals across the cell
 Microbial Nutrition membrane
- Nutrition: process of acquiring nutrients 1. Passive transport: does not require energy,
- Essential nutrients: macronutrients (large quantities: gradient dependent (higher to lower concentration)
proteins, carbs) and micronutrients (small amounts, - Diffusion
enzyme function and maintenance of protein
structure: manganese, zinc)
- Nutrients: inorganic (contains atoms other than C and
H: metals and their salts) and organic (contains C and
H: methane, carbs, proteins, lipids, nucleic acid)
 Sources of essential nutrients - Osmosis
1. Carbon sources
- Organic matter (heterotrophs)
- Carbon dioxide (autotroph)
2. Nitrogen sources
- Nitrogen gas (79% of earth’s atmosphere)
- Part of structure of proteins, DNA, RNA and ATP
3. Oxygen sources
- Oxygen gas (20% of earth’s atmosphere)
- Carbohydrates, lipids, nucleic acids, proteins
- Inorganic salts (phosphates, nitrates) and water
4. Hydrogen sources - Facilitated diffusion
- Organic and inorganic (water, salts, gas) compounds
- Maintain pH, forming H bonds between molecules,
source of free energy in redox reactions of respiration
5. Phosphorus sources
- Phosphate from phosphoric acid found in rocks and
oceanic mineral deposits 2. Active transport
6. Sulfur sources - Active transport
- Rocks, sediments, sulfides, hydrogen sulfide gas
7. Other sources
- Potassium: protein synthesis and membrane function
- Sodium: cell transport
- Calcium: cell wall and endospore stabilizer
- Magnesium: component of chlorophyll, membrane - Group translocation
and ribosome stabilizer
- Iron: component of proteins of cell respiration
- Zinc, copper, nickel, manganese
 - Bulk transport
1.Endocytosis 2.Exocytosis

3.Pinocytosis 4.Phagocytosis

 Environmental factors that influence microbes

1. Temperature
2. Oxygen requirements
3. pH
4. osmotic pressure
 Ecological associations among microorganisms
1. Mutualism
-
2. Commensalism
3. Parasitism
4. Synergism
5. Antagonism
6. Interrelationship between microbes and humans

2. Microbial metabolism
 The basic function of enzymes in cells
 Utilization of energy
 Pathway of bioenergetics