MICROBIOLOGY

WEEK 3

THINGS TO KNOW
KEYWORDS
METABOLISM
ENZYMES
CELLULAR RESPIRATION
FERMENTATION
FISSION
BACTERIAL GROWTH
CULTURE
PLATE COUNTING
SERIAL DILUTION
COMPLEX MEDIA
SELECTIVE MEDIA
DIFFERENTIAL MEDIA
AEROBIC
ANAEROBIC

KEY CONCEPTS
ENZYMES AND METABOLISM
STAGES OF CELLULAR RESPIRATION
AND FERMENTATION
CALCULATING BACTERIAL GROWTH

IDENTIFYING PHYSICAL AND

CHEMICAL REQUIREMENTS FOR
GROWTH
MEDIA TYPES

YOU SHOULD BE ABLE TO

DISTINGUISH BETWEEN PHOTOAUTOTROPH, CHEMOAUTOTROPH, AND
PHOTOHETEROTROPH
DESCRIBE THE ROLE OF ENZYMES IN METABOLISM
DESCRIBE THE PROCESS OF CELLULAR RESPIRATION (GLYCOLYSIS, KREBS
CYCLE, ELECTRON TRANSPORT) AND FERMENTATION
DEFINE MICROBIAL GROWTH, BINARY FISSION AND BUDDING
DESCRIBE METHODS OF MEASURING BACTERIAL GROWTH

CALCULATE GROWTH RATES

IDENTIFY PHYSICAL REQUIREMENTS FOR GROWTH
IDENTIFY NUTRITIONAL REQUIREMENTS FOR GROWTH
DESCRIBE METHODS TO OBTAIN A PURE CULTURE
DIFFERENTIATE BETWEEN CULTURE METHODS INCLUDING SELECTIVE AND
DIFFERENTIAL, ENRICHMENT, ANAEROBIC AND LIVING HOST CELLS
TRANSFER BACTERIA USING ASEPTIC TECHNIQUES

METABOLISM
THE SUM OF ALL CHEMICAL REACTIONS INSIDE OF A CELL = METABOLISM
IS IT DESTRUCTIVE OR CONSTRUCTIVE?
BOTH!
CATABOLISM IS DESTRUCTIVE (BREAKS DOWN BIG MOLECULES INTO SMALLER
MOLECULES, OFTEN TO OBTAIN ENERGY)
ANABOLISM IS CONSTRUCTIVE (BUILDING LARGER MOLECULES OUT OF SMALLER OFTEN
IN ORDER TO STORE ENERGY)
GIVE ME EXAMPLES OF EACH!
CATABOLISM: BREAKING DOWN OF GLUCOSE, LACTOSE, WASTES, ETC.

ANABOLISM: CREATION OF PROTEINS, LIPIDS, CARBOHYDRATES, ETC.

TAKING A CLOSER LOOK AT WORDS

WHAT DO THE FOLLOWING ROOT WORDS MEAN?
CHEMO
CHEMICAL
USES CHEMICAL REACTIONS TO FIX INORGANIC CARBON INTO MOLECULES USEFUL FOR LIFE

PHOTO
LIGHT
USES LIGHT REACTIONS TO FIX INORGANIC CARBON INTO MOLECULES USEFUL FOR LIFE

AUTO
SELF
DEPENDS ONLY ON ITSELF TO PROVIDE THE ENERGY AND BUILDING BLOCKS USEFUL FOR LIFE
(DOES NOT NEED TO INGEST/CONSUME AN INTERMEDIARY)

HETERO
DIFFERENT/OTHER
USES AN INTERMEDIARY TO PROVIDE BUILDING BLOCKS ESSENTIAL FOR MOLECULES NEEDED
FOR LIVING

SO WHAT DO THESE MEAN?

CHEMOAUTOTROPH: USES CHEMICAL REACTIONS TO TURN INORGANIC CARBON (CARBON DIOXIDE) INTO AN
ENERGY SOURCE
BACTERIA LIVING IN/NEAR DEEP SEA VENTS

CHEMOHETEROTROPH: USES CHEMICAL REACTIONS TO TURN INORGANIC CARBON (CARBON DIOXIDE) INTO AN
ENERGY SOURCE OR CONSUMES INTERMEDIARIES TO OBTAIN THE BUILDING BLOCKS TO CREATE MOLECULES
NECESSARY FOR LIFE
FUNGI

PHOTOAUTOTROPH: USES LIGHT REACTIONS TO TURN INORGANIC CARBON (CARBON DIOXIDE) INTO AN
ENERGY SOURCE
PLANTS

PHOTOHETEROTROPH: USES LIGHT REACTIONS TO TURN INORGANIC CARBON (CARBON DIOXIDE) INTO AN
ENERGY SOURCE OR CONSUMES INTERMEDIARIES TO OBTAIN THE BUILDING BLOCKS TO CREATE MOLECULES
NECESSARY FOR LIFE
PURPLE NON-SULFUR BACTERIA

ENZYMES

WHAT ARE ENZYMES MADE OF?

PROTEINS OR RNA (REMEMBER FROM LAST WEEK?)

ENZYME STRUCTURE

After this happens, the enzyme returns to its original state while the substrate is changed.

WHAT HAPPENS TO PROTEIN WHEN YOU

ADD HEAT?

WATCH THIS!
PINEAPPLE & JELL-O

PINEAPPLE & JELL-O EXPERIMENT

EXPLAINED
THE STUDENT TOOK CANNED PINEAPPLE, DRIED PINEAPPLE, FRESH
PINEAPPLE, AND FRESH COOKED PINEAPPLE (2 SECONDS30 SECONDS) AND
TESTED WHETHER OR NOT THE PINEAPPLE VARIABLE IMPACTS THE JELL-OS
ABILITY TO SET
JELL-O IS MADE OF SIMPLE PROTEINS
WHY DID THE STUDENT GET THE RESULTS THAT HE OBSERVED?
PINEAPPLE CONTAINS AN ENZYME INTENDED TO BREAK DOWN PROTEINS
BUT WHY DIDNT ALL OF THE PINEAPPLE HE TESTED HAVE THE SAME RESULT?
BECAUSE THE ENZYMES WERE RENDERED USELESS THROUGH BOILING (HEAT + PROTEIN =
CHANGE IN PROTEIN STRUCTURE)

CAN ANYTHING IMPEDE AN ENZYME

WITHOUT PERMANENTLY DAMAGING IT?
COMPETITIVE INHIBITION

NONCOMPETITIVE INHIBITION

THINK ABOUT IT
WHY WOULD BACTERIAL TOXINS IN THE FORM OF ENZYMES BE DAMAGING TO
OUR CELLS?

TIME FOR SOME DEFINITIONS!

OBLIGATE
REQUIRED

FACULTATIVE
CAN MAKE USE OF

AEROBE
OXYGEN

ANAEROBE
NO OXYGEN

THE CIRCLE OF LIFE

CELLULAR RESPIRATION
FIRST, LETS WATCH THIS, THEN WELL SLOW IT DOWN AND REVIEW

RESPIRATION STEPS
If oxygen
present

Krebs cycle
ETC

In
absence
of
oxygen

fermentation

glycolysis

GLYCOLYSIS

REBS CYCLE (OXYGEN ONLY)

Products:
3 NADH
1 GTP
1 FADH2
And releases 2 Co2

ELECTRON TRANSPORT CHAIN (ETC)

High energy molecules formed in KREBS cycle and glycolysis are used to fuel ATP pump and make water from
oxygen!

ERMENTATION (ABSENCE OF OXYGEN)

Very few ATP produced during fermentation. Why then do organisms undergo fermentation at all?

BACTERIAL GROWTH
REFERS TO THE NUMBER
OF BACTERIA, NOT THE SIZE

PHASES EXPLAINED
DURING LAG PHASE, INDIVIDUAL BACTERIA ARE GROWING AND STORING ATP

EVENTUALLY, DURING THE EXPONENTIAL PHASE, THE BACTERIA ARE

DOUBLING IN SIZE
MOST BACTERIA HAVE A DOUBLING TIME BETWEEN 20 MINUTES AND 20 HOURS

DURING THE STATIONARY PHASE THE AMOUNT OF NUTRIENTS IS LIMITING

AND/OR BACTERIAL WASTES ARE LIMITING NEW GROWTH.
GROWTH AND DEATH OCCURRING AT SAME RATE

EVENTUALLY DEATH RESULTS FROM STAGNATION

THIS HAPPENS FASTER THAN GROWTH!

BINARY FISSION

STOP, CALCULATE AND LISTEN

FIND A PARTNER, GO!

E.COLI HAS A DOUBLING TIME OF 20 MINUTES

HOW MANY CELLS WILL THERE BE IF WE INOCULATE A BROTH WITH PLENTIFUL
NUTRIENTS WITH ONE CELL OF E. COLI
AFTER 20 MINUTES?
AFTER 1 HOUR?
.AFTER 3 HOURS?

E.COLI GROWTH

time

20

40

60

80

100 120 140 160 180

# cells

16

32

64

128 256 512

INOCULATION
DO WE EVER JUST INOCULATE WITH ONE CELL?

SERIAL DILUTION

PLATE COUNTS

TYPES OF MEDIUM FOR MICROBIAL GROWTH

SYNTHETIC: MADE CHEMICALLY
COMPLEX: ONE WHERE EXACT INGREDIENTS ARE NOT KNOWN
SELECTIVE: SELECTS FOR THE GROWTH OF ONE PARTICULAR TYPE OF
MICROORGANISM
DIFFERENTIAL: PROVIDES A MEANS OF DETECTING BETWEEN DIFFERENT
TYPES OF GROWTH ON THE PLATE
ENRICHMENT: PROVIDES A NUTRIENT THAT ONE TYPE OF BACTERIA NEEDS,
THUS PROVIDING FOR GROWTH OF THE ONE AND NOT OTHER TYPES

WHAT MEDIUM WOULD YOU

CHOOSE FOR GROWTH
OBLIGATE ANAEROBE
FACULTATIVE ANAEROBE

WHAT TYPE OF GROWTH WOULD YOU

EXPECT IN A BROTH FOR THE FOLLOWING?
OBLIGATE ANAEROBE
FACULTATIVE ANAEROBE
OBLIGATE AEROBE
MICROAEROPHILE (WHAT?!)

TECHNIQUE

Frequent subculturing and aseptic technique!

A CLOSER LOOK