EXAM 2 DEFINITIONS & MICROBES

Biochemical FERMENTATION

internally balanced anaerobic oxidation-reduction process in which fermentable substrate becomes oxidized and the METABOLITE is REDUCED

Also NAD+ has to be regenerated

(NADH — oxidized —> NAD+)

(OXIDIZED PRODUCT = REDUCED PRODUCT)

Energy obtained via substrate- level phosphoralatin

Substrate-level phosphorylation

When give away a phosphate group from some high energy compound and put on ADP to form ATP or GTP

ATP

Adenosine triphosphate

Main source of energy for synthesis of complex macromolecules

GTP

guanosine triphosphate

Can be used to transport energy but mostly a signal molecule

Catabolism

energy yielding, degradative porcess

WHAT are the two mechanisms of ATP production

substrate level phosphorylation (fermentation anaerobic)

Proton motive force (respiration)

Anabolism

Mechanism that takes smaller untis like nutrients, cells or amino acids and bonds them together to create bigger structures

amino acids

5 MAIN pathways of glycolysis

Embden-Meyerhof-Parnas (EMP) pathway

Entner-Doudoroff pathway

Phosphoketolase pathway

Pentose phosphate pathway

what is glycolysis

it is the breakdown of glucose.

It metabolises glucose to pyruvate

And provides ATP (through susbtrate-level phosphoralation) and

Reduces coenzymes (ex: NADH)

COENZYME

Non-protein, organic molecule that partheners with and helps enzymes

Specific type of organic cofactor

Coenzymes facilitate chemical reactions

Embden-Meyerhof-Parnas (EMP) pathway

= fructose-1, 6-biosphosphate aldolase pathway

It is the most common route and found in all major groups of organisms

Entner-Doudoroff pathway

= ketogluconate pathway

And it is the glucose degradation and fermentation to ethanol by Zymomonas mobilis

Phosphoketolase pathway

= hexose monophosphate pathway

Major route in many HETEROFERMENTATIVE organisms ex: Leuconostoc

Pentose phosphate pathway

both catabolic and anabolic roles

Main difference between the Embden-Meyerhof-Parnas (EMP) pathway and Enter-Doudoroff pathway

Steps in the Embden-Meyerhof-Parnas (EMP) pathway

1. Preparatory reaction (glucose → fructose 1,6-bisphosphate (MAIN INTERMADIATE) → Glyeraldehyde 3-phosphate)

2. Glyceraldehyde 3-phosphate → Pyruvate and ATP

   1. SUBSTRATE-LEVEL PHOSPHORALATION

3. Pyruvate

Overall reaction of the Embden-Meyerhof-Parnas (EMP) pathway

C₆H₁₂O₆ + 2 NAD + 2 ATP + 2 P_i (inorganic phosphate) → 2 C₃H₄O₃ + 2 NADH + 2H⁺ + 4 ATP

1GLUCOSE → 2 pyruvate

2 ATP → 4ATP; net 2 ATP

NAD+ —REDUCED—> NADH

main steps of the Entner-Doudoroff pathway

1. Glucose → 2-keto-3-deoxy-6-phosphogluconate (MAIN intermediate) → PYRUVATE + Glyceraldehyde 3-phosphate

2. Glyceraldehyde 3-phosphate → PYRUVATE + ATP

2 PYRUVATE

2 ATP

NAD+ —REDUCED—> NADH

Main steps phosphoketolase pathway

found in HETEROFORMATIVE LACTIC ACID BACTERIA

GLUCOSE → Xylulose 5-P

Xylulose 5-P + Pi —phosphoketolase (enzyme)→ ETHANOL

Xylose 5-P → Pyruvate → LACTATE

NADH —OXIDIZED—> NAD+

KEY THINGS ABOUT pentose phosphate pathway

• found in many bacteria and most eukaryotic organisms

• Both anabolic and catabolic roles

• Important in the provision of NADPH

which glycolytic pathway does ETHANOLIC fermentation use

Embden-Meyerhof (EMP)

Ethanolic fermentation main products

EtOH + CO2

which glycolytic pathway does Homolactic acid fermentation use

Embden-Meyerhof

main products of homolactic acid fermentation

lactic acid

which glycolytic pathway does HETEROLACTIC ACID fermentation use

Entner-Doudoroff

Main products of HETEROLACTIC ACID FERMENTATION

lactic acid + EtOH + CO2

which glycolytic pathway does PROPIONIC ACID fermentation use

Embden-Meyerhof (EMB)

Main products of PROPIONIC ACID FEREMNTATION

Propionic acid + CO2

which glycolytic pathway does MIXED ACID fermentation use

Embden-Meyerhof (EMB)

MAIN PRODUCTS OF MIXED ACID FERMENTATION

EtOh + acetic acid + lactic acid + succinic acid + formaic acid + H2 + CO2

which glycolytic pathway does Butyric acid fermentation use

Embden-Meyerhof

main products of BUTYRIC ACID fermentation

butyric acid + butanol + acetone + CO2

which glycolytic pathway does BUTANEDIOL fermentation use

Embden-Meyerhof (EMB)

main products of butanediol fermentation

butanediol + CO2

what industrial product is made through __________ + _____ fermentation

When a terminal electron acceptor is available what is the alternative to fermentation

TCA cycle and respiration

Pyruvate → acetate —TCA cycle—> respiration

TCA cycle purpose

When terminal acceptro is available, TCA occurs as an alternative to fermentaiton —lead to—> respiration

TCA intermediates serve as building blocks for amino acids, nucleotides and lipids

PRIMARY source of NADH and FADH₂ for aerobic respiration (they are elctron carriers needed for electron transport chain, creating proton motive force that drive ATP synthesis)

What is the PASTEUR EFFECT

• cells that switch from fermentation to respiration consume glucose more slowly brcause respiration yields more ATP per glucose

• (WHAT IS THE INHIBITOR THING… TOO MUCH ATP INHIBIT MITOCHONDRIA

importance of Oxaloacetate in TCA cycle

its concentration directly control cycle flux

If it is depleted, citrate synthase cannot function and entire cycle SLOWS or stops

What are the major industrial products obtained from microbes

1. Fermented foods/bbeverages

2. Flavoring agents & food supplements

3. Organic acids

4. Solvents

5. Enzymes and microbial transformations

6. Inhibitors

7. Products from genetically engineered micrboes

Industrial products obtained from microbes can be…

Microbial cells (the microbial cells themselves)

or cell products

• products made by the cell

  • Enzymatic biotransformation products (products produced by an enzyme in the cell)

  • Fermentation products

Oxychemicals obtained by microbial fermentation

are organic chemical compounds containing oxygen that are produced on a commercial scale through metabolic activity of microorganisms

primary metabolite

It is produced during active growth

It is generally a consequence of energy metabolism and necessary for the CONTINUED growth of the microbe

secondary metabolite

it is produced AFTER the growth phase nears completion

The product is formed by metabolism of an intermediate of PRIMARY METABOLISM

Trophophase

In industrial microbiology, It is what the growth phase of microbes is called

• at this stage- Primary metabolites are produced

idiophase

it is the phase involved in the production of metabolites (idiolites)

Follows the trophosphase

It is a shift in the microorganism’s metabolic activity

Begin to produce secondary metabolites

Upstream process

at this stage, fermentation of raw products occurs. This also entails media development:

• Ensuring optimal conditions

• Source of trace minerals

• N, P, C sources

• Aeration

• PH

• T

Also part of the process:

• improve strain and production of strain

Downstream process

purifying and recovery of product after industrial fermentation

To keep in mind :

• yield at each step

• Concentration and stability of product

• Cost

• Purity requirement → methods

Aeration

aerobic v.s anaerobic process

It is how control oxygen in fermentation system

• how to do in large-scale vessles, how will mix

Important aspect to keep in mind regarding Temperature when utilizing fermentors for large scale industrial fermentation

Temperature will increase as microorganisms grow

Transfer of energy NOT 100% efficient → some of the energy will be released as heat (waste)

Need cooling coils and water jackets to maintain optimal temperature

airlift fermentor

Chemostat

Type of continuous fermentation where the rate of new medium fed into the fermentor is proportional to the rate of cell loss by the outflow of it to achieve steady state

Turbidostat

A method of continuous fermentation where cell growth is kept constant by using turbidity to monitor the biomass concentration, and the rate of feed of nutrient solution i adjusted to maintain a constant turbidity

steady state

Cell loss is balanced by cell growth

D = mu

Dilution factor = Growth rate

What are the main principles for separating fermentation products from solution

size

Diffusion

Charge

Vapor pressure

Solubility

Surface tension

Weight

what are the maiin principles for opening cells to isolate microbila metabolites

• physical methods

  • Mills

  • Freeze/thaw

• Chemical methods

  • Lyophilization

  • Treatment with acid

• Biological methods

  • Cell wall digesting enzymes

  • Agents that inhibit cell wall synthesis

freeze/thaw

Methods for purifying microbial metabolites