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METABOLIC PATHWAYS
ALLOWS CELLS TO USE METABOLITES TO THEIR FULL
POTENTIAL
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METABOLISM
THE SUM OF ALL CELLULAR REACTIONS
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TYPES OF METABOLIC REACTIONS
ANABOLIC VS CATABOLIC
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CATABOLISM
AEROBIC VS ANAEROBIC
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COUPLING AGENTS
ATP
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ATP STRUCTURE
• ADENINE
• RIBOSE
• THREE PHOSPHATE GROUPS
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THE IMPORTANCE OF HIGH ENERGY BONDS
• THIRD PHOSPHATE BOND
• UNSTABLE
• HYDROLYSIS IS EXERGONINC
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ENERGY METABOLISM
OXIDATION VS REDUCTION
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BIOLOGICAL OXIDATION
• REMOVAL OF ELECTIONS IN INORGANIC MOLECULES
• REMOVAL OF HYDROGENS IN ORGANIC MOLECULES
• DEHYDROGENASES
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BIOLOGICAL REDUCTIONS
• ADDITIONS OF ELECTRONS IN INORGANIC
• ADDITIONS OF HYDROGENS IN ORGANIC
• OXIDATION AND REDUCTION MUST OCCUR SIMULTANEOUSLY
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COUPLING AGENTS
COENZYMES
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SUBSTRATES FOR OXIDATION
• INORGANIC COMPOUNDS (REDUCED IRON, SULFUR OR NITROGEN)
• HEXOSES (FRUCTOSE, GLUCOSE, LACTOSE)
• POLYSACCHARIDES
• PROTEINS
• FATS
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GLUCOSE
• HEXOSE• MAIN SUGAR IN
VERTEBRATES• SUGAR RELEASED FROM
STARCH IN PLANTS• CONSTITUENT OF
SUCROSE • BEGINNING POINT FOR
EMBDEN MYERHOFF, ENTNER DUODOROFF, PENTOSE PHOSPHATE PATHWAYS
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THE IMPORTANCE OF ELECTRON ACCEPTORS
• OXYGEN
• OTHER INORGANIC MOLECULES
• ORGANIC MOLECULES
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AEROBIC RESPIRATION VS FERMENTATION
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AEROBIC RESPIRATION
• NEED ELECTRON ACCEPTOR FOR OXIDATIONS TO OCCUR
• MOST USE OXYGEN
• ACCESS TO FULL ENERGY OF GLUCOSE REQUIRES OXIDATION
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FERMENTATION
• MOST ORGANISMS CAN EXTRACT ENERGY BY EMBDEN MYERHOFF PATHWAY/GLYCOLYSIS
• DOES NOT REQUIRE OXYGEN• ELECTRONS ARE GIVEN BACK TO AN
ORGANIC MOLECULE THAT IS PYRUVATE OR A DERIVATIVE OF PYRUVATE
• LACTATE, ALCOHOL, ACIDS, ETC
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ANAEROBIC RESPIRATION
• USES FINAL INORGANIC ELECTRON ACCEPTOR OTHER THAN ATMOSPHERIC OXYGEN
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CLASSIFICATION OF ORGANISMS BY THEIR USE OF OXYGEN
• STRICT OR OBLIGATE AEROBES
• STRICT OR OBLIGATE ANAEROBES
• FACULATIVE ANAEROBES
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EMBDEN - MEYERHOFF PATHWAY
GLYCOLYSIS
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EMBDEN-MEYERHOFF
• TEN STEP PATHWAY
• FOUND IN BOTH AEROBIC AND ANAEROBIC ORGANSIMS
• PYRUVATE IS THE END PRODUCT
• CAN BE REDUCED THROUGH FERMENTATION
• OR OXIDIZED FURTHER
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PRODUCTS OF EMBDEN MEYERHOFF
• NADH
• ATP
• PYRUVATE
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PHASES OF THE EMBDEN MEYERHOFF PATHWAY
• PREPRATORY AND CLEAVAGE– STEPS 1-5
• OXIDATIVE – STEPS 6 & 7
• ATP GENERATING PHASE– STEPS 8-10
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PREPARATION AND CLEAVAGE
• FRUCTOSE 1,6 BISPHOSPHATE
• TWO TERMINAL PHOSPHATE GROUPS
• DHAP AND GLYCERALDEHYDE 3 PHOSPHATE
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DHAP AND GLYCERALDEHYDE 3 PHOSPHATE
• DHAP = KETOSE
• GLYCERALDEHYDE 3 PHOSPHATE = ALDOSE
• ONLY GLYCERALDEHYDE 3 PHOSPHATE OXIDIZED
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OXIDATION AND ATP GENERATION
• NEGATIVE ENERGY YIELD SO FAR
• ATP PRODUCTION LINKED TO OXIDATION
• GLYCERALDEHYDE 3 DEHYDROGENASE
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PYRUVATE & ATP FORMATION
• ISOMERIZATION REACTION TO INCREASE AMOUNT OF FREE ENERGY
• 3 PHOSPHOGLCERAATE TO 2PHOSPHOGLYCERATE TO PHOSPHENOLPYRUVATE
• PHOSPHOENOLPYRUVATE TO PYRUVATE
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IMPORTANCE OF PYRUVATE
• BRANCHING POINT
• FATE DEPENDS ON ORGANISM AND OXYGEN
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CATABOLISM OF GLUCOSE BY CELLULAR RESPIRATION
ENERGY RELEASE IS EXACTLY THE SAME AS WHEN SUGAR IS
THROWN ONTO A FIRE
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BURNING GLUCOSE IN OXYGEN RELEASES 686,00
CALORIES PER MOLE
CELLS WILL NOT CAPTURE ALL THIS ENERGY
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THE CATABOLISM OF GLUCOSE
• GLYCOLYSIS—EMBDEN MERYEHOFF PATHWAY
• KREBS CYCLE—TCA CYCLE
• ELECTRON TRANSPORT
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STEP ONE OF GLYCOLYSIS
• SUBSTRATE PHOSPHORYLATION OF GLUCOSE
• CONVERSION TO GLUCOSE 6 PHOSPHATE
• GLUCOSE TRAPPING
• HEXOKINASE IS THE ENZYME USED IN EUKARYOTES
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HORMONAL REGULATION
• MAMMALS REGULATE THIS PHOSPHORYLATION WITH THE HORMONE INSULIN
• THERE ARE MANY HORMONES THAT HELP REGULATE SUGAR METABOLISM
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STEP TWO OF GLYCOLYSIS
• GLUCOSE 6 PHOSPHATE IS REARRANGED INTO FRUCTOSE 6 PHOSPHATE
• PHOSPHOFRUCTOKINASE IS THE ENZYME USED
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STEP THREE OF GLYCOLYSIS
• SUBSTRATE PHOSPHORYLATION OF FRUCTOSE 6 PHOSPHATE TO FORM FRUCTOSE 1, 6 DIPHOSPHATE
• CAN ALSO BE CALLED FRUCTOSE 1,6 BISPHOSPHATE
• PHOSPHOFRUCTOKINASE IS THE ENZYME USED
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IMPORTANCE OF PHOSPHOFRUCTOKINASE
• EXTENSIVELY STUDIED ENZYME
• KEY REGULATOR OF GLYCOLYTIC PATHWAY
• ACTIVITES STIMULATED BY FRUCTOSE 6 PHOSPHATE, AMP AND ADP
• ACTIVITIES INHIBITED BY ATP AND CITRATE
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PHOSPHOFRUCTOKINASE ALLOWS THE CELL TO
BALANCE THE RATES OF GLYCOLYSIS AND THE
KREBS CYCLE
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STEP FOUR OF GLYCOLYSIS
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STEP FIVE OF GLYCOLYSIS
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NO ENERGY HAS BEEN PRODUCED
• IN FACT WE HAVE INVESTED TWO ATPs
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STEP SIX OF GLYCOLYSIS
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STEP SEVEN OF GLYCOLYSIS
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STEP EIGHT OF GLYCOLYSIS
• THIS PRIMES THE GLUCOSE FOR ATP PRODUCTION
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STEP NINE OF GLYCOLYSIS
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STEP TEN OF GLYCOLYSIS
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FOUR ATP MOLECULES WILL BE HARVESTED IN THIS PORTION OF
THE PATHWAY
• TWO ATPs INVESTED EARLIER MUST BE PAID BACK
• THE NET YIELD OF GLYCOLYSIS IS 2 ATPs
• TWO NADHs
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AEROBIC PATHWAYS
• TCA CYCLE/KREBS CYCLE
• ELECTRON TRANSPORT SYSTEM– PRODUCE CARBON DIOXIDE
• WATER
• ENERGY– STORED AS ATP
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KREBS CYCLE
TCA CYCLE
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PREPRATORY RX
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CITRIC ACID AS A REGULATOR OF PHOSPHOFRUCTOKINASE
ACTIVITY
• CITRIC ACID IS AN INHIBITOR OF PHOSPHOFRUCTOKINASE
• PREVENTS GLYCOLYSIS FROM OCCURRING WHEN IT IS NOT NECESSARY
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CITRATE IS A TERTIARY ALCOHOL
• MUST BE CONVERTED TO A SECONDARY ALCOHOL
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ELECTRON TRANSPORT SYSTEM
• GLYCOLYSIS = 2 ATPs
• TCA = 2 ATPs
• MOST OF ATPs PRODUCED BY ETS
• SERIES OF ELECTRON CARRIERS
• REDOX REACTIONS
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ELECTRON TRANSPORT SYSTEM
• FROM MORE NEGATIVE REDUCTION POTENTIALS TO MORE POSITIVE POTENTIALS
• DIFFERENCE BETWEEN NADH AND OXYGEN IS ABOUT 1.14 VOLTS
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SITES OF ATP SYNTHESIS
• BETWEEN NADH AND COENZYME Q
• BETWEEN CYTOCHROME B AND C1
• BETWEEN CYTOCHROME A AND OXYGEN
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ELECTRON TRANSPORT SYSTEM
• ALLOWS SMALL RELEASES OF ENERGY IN SMALL STEPS
• ESTABLISHES PROTON AND ELECTRICAL GRADIENT
• FOUND IN INNER MITOCHONDRIAL MEMBRANE• FOUR COMPLEXES OF CARRIERS• CYTOCHROME C AND COENZYME Q CONNECT
COMPLEXES
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OXIDATIVE PHOSPHORYLATION
• AS MANY AS THREE ATPs FOR EACH TWO ELECTRONS FROM NADH TO OXYGEN
• AS MANY AS TWO ATPs FOR EACH TWO ELECTRONS FROM FADH2 TO OXYGEN
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BACTERIAL ELECTRON TRANSPORT SYSTEMS
• SOME RESEMBLE MITOCHONDRIAL CHAINS
• MANY ARE VERY DIFFERENT
• VARIOUS TYPES OF CYTOCHROMES
• SOME HIGHLY BRANCHED
• MAY BE SHORTER
• HAVE LESS ATP PRODUCTION
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OXIDATIVE PHOSPHORYLATION
• CHEMIOSMOSIS
• CONFORMATIONAL CHANGE HYPOTHESIS
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CHEMIOSMOSIS
• PROTONS MOVE INTO INTERMEMBRANAL SPACE
• CAUSES PROTON MOTIVE FORCE• GRADIENT OF PROTONS• MEMBRANE POTENTIAL DUE TO
UNEVEN DISTRIBUTION OF CHARGES• ATPs MADE AS H+ DIFFUSE BACK
INTO MATRIX
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OTHER USES OF PROTON MOTIVE FORCE
• TRANSPORT ACROSS MEMBRANES
• ROTATION OF BACTERIAL FLAGELLA
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CONFORMATIONAL CHANGE HYPOTHESIS
• ENERGY RELEASED CAUSES CHANGES IN THE SHAPE OF ATPase ENZYME
• CHANGES DO ACTUALLY OCCUR DURING ELECTRON TRANSPORT
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INHIBITORS OF ELECTRON TRANSPORT
• BLOCK ELECTRON TRANSPORT
• UNCOUPLE ATPase AND ELECTRON TRANSPORT
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BLOCKERS
• PERICIDIN -- COMPETES WITH COENZYME Q FOR ELECTRONS
• ANTIMYCIN A -- BLOCKS ELECTRON TRANSPORT BETWEEN B AND C
• CYANIDE & AZIDE -- STOP TRANSFER BETWEEN CYTOCHROME A AND OXYGEN
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UNCOUPLERS
• DO NOT AFFECT ELECTRON TRANSPORT
• DISCONNECT OXIDATIVE PHOSPHORYLATION FROM ELECTRON TRANSPORT
• ENERGY IS RELEASED AS HEAT
• DINTITOPHENOL & VALINOMYCIN
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ATP YIELDS
GLYCOLYSIS & AEROBIC RESPIRATION
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GLYCOLYSIS
• NET YIELD OF 2 ATPs
• TWO NADHs PRODUCE SIX ATPs
• TOTAL OF EIGHT ATPs
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PREPARATORY REACTION
• 2 NADHs FOR A YIELD OF 6 ATPs
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TCA CYCLE
• 2 ATPs
• 6 NADHs FOR 18 ATPs
• 2 FADH2s FOR 4 ATPs
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ANAEROBIC PATHWAYS
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LACTATE FERMENTATION
• DIRECT TRANSFER FROM NADH TO PYRUVATE
• LACTATE DEHYDROGENASE
• MAJOR PATHWAY IN MANY ANAEROBIC BACTERIA
• MUSCLES
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ETHANOL FERMENTATION
• NADH AND ACETALDEHYDE (A DERIVATIVE OF PYRUVATE)
• TWO STEP SEQUENCE• DECARBOXYLATION--
PYRUVATE DECARBOXYLASE
• ACETALDEHYDE REDUCTION -- ALCOHOL DEHYDROGENASE
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TYPES OF FERMENTATIONS
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ALCOHOLIC FERMENTATION
• FUNGI
• ALGAE
• BACTERIA
• PROTOZOA
• SUGARS --------> ALCOHOL
• ALCOHOL DEHYDROGENASE
• NADH DONOR
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LACTIC ACID FERMENTATION
• REDUCTION OF PYRUVATE TO LACTATE• MORE COMMON THAN ALCOHOL
FERMENTATION• BACTERIA• ALGAE• WATER MOLDS• PROTOZOA• ANIMAL MUSCLE CELLS
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HOMOLACTIC ACID FERMENTERS
• LACTATE DEHYDROGENASE
• USE GLYCOLYTIC PATHWAY
• TWO KINDS OF LACTIC ACID FERMENTERS
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HETEROLACTIC FERMENTERS
• MANY PRODUCE LACTATE
• ETHANOL
• VARIETY OF OTHER PRODUCTS
• USES PHOSPHOKETALOSE PATHWAY
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FORMIC ACID FERMENTERS
MIXED ACID FERMENTATION
BUTANEDIOL FERMENTATION
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MIXED ACID FERMENTATION
• ENTEROBACTERIACEAE
• PYRUVATE ---->
• HYDROGEN GAS
• CARBON DIOXIDE
• FORMIC HYDROGENLYASE
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MIXED ACID FERMENTATION
• ETHANOL
• COMPLEX MIXTURE OF ACIDS
• ACETIC
• LACTIC
• SUCCINIC
• FORMIC
• ESCHERICHIA, SALMONELLA & ETC
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ATP GENERATION IN FORMIC ACID
FERMENTATION• SOMETIME GENERATE ATP WILL
REOXIDIZING NADH
• USE ACETYL CO A -------> ACETYL-PHOSPHATE
• DONATES TO ADP
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FERMENTERS OF AMINO ACIDS
• STICKLAND REACTION
• CLOSTRIDIUM SPECIES
• GENERATE AMMONIA, HYDROGEN SULFIDE, FATTY ACIDS AND AMINES
• PUTREFACTION ODORS
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ANAEROBIC RESPIRATION
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ANAEROBIC RESPIRATION
• ELECTRON TRANSPORT SYSTEMS DONATE ELECTRONS TO INORGANIC MOLECULES OTHER THAN OXYGEN
• NITRATE
• SULFATE
• CARBON DIOXIDE
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NITRATE
• NO3 + 2 e- + 2 H+ -------> NO2 + H2O
• NITRATE IS THE ELECTRON ACCEPTOR
• NITRATE REDUCTASE REPLACES CYTOCHROME OXIDASE
• NOT EFFECTIVE
• PRODUCT TOXIC
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DENITRIFICATION
• REDUCED ALL THE WAY TO N2
• ACCEPTS FIVE ELECTRONS
• IS NONTOXIC
• 2 NO3 + 10 e- + 12 H+ ------> N2 + 6 H2O
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THE PENTOSE-PHOSPHATE PATHWAY
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THE PENTOSE PHOSPHATE PATHWAY
• HEXOSE MONOPHOSPHATE PATHWAY
• CAN OPERATE AT SAME TIME AS EMBDEN-MEYERHOF
• AEROBIC OR ANAEROBIC• PROVIDES THREE TO SEVEN
CARBON SUGARS PHOSPHATE• NADPH IS ALSO PRODUCED
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END PRODUCTS
• FRUCTOSE 6 PHOSPHATE
• GLYCERALDEHYDE 3 PHOSPHATE
• THREE CARBON DIOXIDE MOLECULES
• 6 NADPHs
• INTERMEDIATES FOR ANABOLIC REACTIONS
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P GLUCOSE 6 - PHOSPHATE
`
NADPH
6-PHOSPHOGLUCONATE
NADPH
RIBULOSE 5 - PHOSPHATE
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END PRODUCTS
ERYTHROSE 4 PHOSPHATE XYLOSE 5 PHOSPHATE
GLYCERALDEHYDE 3 PHOSPHATE FRUCTOSE 6 PHOSPHATE
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IMPORTANCE OF PENTOSE PHOSPHATE PATHWAY
• MAKES ATP• NADPH ACTS AS SOURCE OF
ELECTRONS FOR BIOSYNTHESIS• PRODUCES FOUR AND FIVE CARBON
SUGARS WHICH CAN BE USED • PRODUCES HEXOSE SUGARS FOR
MICROBES GROWN ON PENTOSE CARBON SOURCE
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ENTNER DOUDOROFF PATHWAY
• STARTS WITH GLUCOSE 6 PHOSPHATE
• DEHYDRATION FORMS KDPG
• END PRODUCTS ARE PGAL AND PYRUVATE
• PGAL CAN ENTER LOWER PART OF GLYCOLYSIS
• ONE ATP, ONE NADPH AND ONE NAD
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BETA OXIDATION
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BETA OXIDATION
• REMOVAL OF TWO CARBON GROUPS
• TO FORM ACETYL CoA– MAY ENTER KREBS CYCLE
• NADH AND FADH2 IS FORMED
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PROTEIN METABOLISM
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AMINO ACID BIOSYNTHESIS
• ADD AMINE GROUP TO KETO ACID
• SIMPLE ONE STEP TRANSMINATION REACTION– GLUTAMATE AND ASPARTATE ARE
EXAMPLES
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CATABOLISM OF AMINO ACIDS
• REMOVAL OF AMINE GROUP FROM AMINO ACID
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•
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CATABOLISM OF PROTEINS AND AMINO ACIDS
• PATHOGENIC, FOOD SPOILERS, SOIL MICROBES
• PROTEASE ENZYMES• DEAMINATION• TRANSAMINATION• ALPHA KETO ACID ACCEPTOR• AMMONIUM ION MAY MAKE
MEDIUM ALKALINE