Midterm Flashcards
catalysts for biological reactions
enzymes
Enzymes can become inactive when they coagulate during:
Heating Treatment with: alcohol salts of heavy metals acids and bases
inorganic or organic substance which speeds up the rate of a chemical reaction without itself entering the reaction
Catalyst
organic catalysts made of protein
Enzymes
How do enzymes catalyze?
They lower the activation energy needed to start the chemical reaction
Enzymes begin to be destroyed at:
above 45 degrees Celsius
The enzyme must form a _________ with the substance or substances whose reaction rate it affects
temporary association
The association between enzyme and substrate is thought to form a close physical association between the molecules and is called-
enzyme-substrate complex
While the enzyme-substrate complex is formed,
enzyme action takes place
Upon completion of the reaction, the enzyme and product(s)-
separate
molecules upon which an enzyme acts
substrate
The enzyme is shaped so that it can only lock up with-
a specific substrate molecule.
Each enzyme is specific for
one and only one substrate
Source of catalase
potato
Role of catalase
Decomposition of hydrogen peroxide
Source of glutatione
animal cells
Role of glutathione
Protects red cells from oxidation damage
Source of oxidase
living cells
Role of oxidase
Oxidizes carbohydrates, fats and proteins
Enzymes end in
-ase
Enzymes that identify reacting substance:
sucrase-reacts sucrose
lipase - reacts lipid
Enzymes that describe function of enzyme
oxidase-catalyze oxidation
hydrolase - catalyze hydrolysis
Common names of digestion enzymes use:
-in
Examples of digestion enzymes
pepsin, trypsin, chymotrypsin
Classifications of Enzymes
Oxidoreductases Transferases Hydrolases Lyases Isomerases Ligases
Reaction catalyzed by oxidoreductases
oxidation-reduction
Reaction catalyzed by Transferases
transfer group of atoms
movement of a functional group from one molecule to another
Reaction catalyzed by Hydrolases
hydrolysis
Reaction catalyzed by Lyases
add/remove atoms to/from a double bond
Reaction catalyzed by Isomerases
rearrange atoms
Reaction catalyzed by Ligases
combine molecules using ATP
LER
reducing agent (redox)
GEO
oxidizing agent
basic reaction of Transferases
AX + B ——> A + BX
Lyases catalyze the cleavage of:
C-C, C-O,C-S and C-N bonds by any other means than hydrolysis or oxidation
- the reduction of methylene blue to methylene white by formaldehyde is rapidly catalyzed by fresh milk but not by boiled milk
- an example of oxidation in the absence of O2 with an organic hydrogen acceptor
Schardinger Reaction
catalyzing enzyme present in Schardinger reaction
xanthine oxidase
organic hydrogen acceptor in Schardinger reaction
methylene blue
Reactants in TT1
Boil –> Methylene Blue –> mix –> paraffin oil –> water bath
Reactants in TT2
Methylene Blue –> formalin –> mix –> paraffin oil –> water bath
Reactants in TT3 (Control)
Methylene blue –> mix –> paraffin oil –> water bath
Reaction in TT1
Light blue
did not decolorize after heating
Reaction in TT2
Light blue
decolorizes after to white heating
Reaction in TT3 (Control)
Light blue
decolorizes to white after heating
- is a compound containing three amino acids
- an antioxidant in the body
- vital component in the destruction of free radicals
- detoxification of harmful substances within the body
glutatione
three amino acids in glutathione
glutamate
cysteine
glycine
- Test used for detecting the presence of free thiol groups of cysteine in proteins
- used for the identification of ketones in urine testing
Nitroprusside Reaction
The color that indicates positive result in Nitroprusside Test
red
Factors that affect the speed of decomposition of hydrogen peroxide by catalase:
concentration of catalase
concentration of hydrogen peroxide
hydrogen ion concentration
temperature
reaction involved in liver catalase
absorption of hydrogen peroxide at the catalase surface
The optimum pH of catalytic decomposition of H2O2
6.8 - 7.0
Reaction in liver catalase
Liver + H2O ——> H2O + O2 (bubbles)
Roles of catalase in liver
- common antioxidant enzyme
- helps body break down H2O2, thus prevents accumulation of CO2 bubbles in the blood
- very potent enzyme; can decompose millions of H2O2
- uses H2O2 to oxidize harmful toxins
Potentially harmful toxins oxidized by H2O2
formaldehyde
formic acid
alcohol
phenol
chemicals used in Nitroprusside test
sodium nitroprusside
ammonium hydroxide
the substance added to the mixture of milk and methylene blue to prevent oxygen from reacting with oxidase
paraffin oil
The compound responsible for the rapid reduction of methylene blue to methylene white in the presence of the enzyme present in milk
formaldehyde
It is a group present in proteins that is being liberated in Nitroprusside Reaction for Glutathione to indicate a positive result
thiol
It is an enzyme from potato responsible for the decomposition of hydrogen peroxide
catalase
Source of catalase in animal enzyme activity
liver
Source of specimen for vegetable/fruit oxidase and catalase:
potato
guava
chico
apples
oxidizes carbohydrates, fats and proteins (main source of body energy)
found in living cells
Oxidase
- a biochemical process in which fruit or vegetable tissues turn brown when exposed to oxygen
- one of the most important color reactions that affects fruits, vegetables and seafood
- can adversely affect flavor and nutritional value
Enzymatic browning
Enzymatic browning is catalyzed by:
polyphenol oxidase or phenolase
Enzymatic browning takes place at:
warm temperature
pH 5.0 - 7.0
Enzymatic browning can economically benefit:
tea, coffee, cocoa, chico
Enzymatic browning can economically jeopardize:
fruits and vegetables
in the presence of oxygen from the air, this enzyme catalyzes the formation of brown pigments called melanin
phenolase
Ways to reduce enzymatic browning
- Treatment of slices with organic substances such as ascorbic acid, citric acid or acetic acid
- Soaking in plain water
- Heating
- acts as an antioxidant
- oxygen preferentially reacts with this rather than phenolic compounds in the fruit or vegetable
- browning does not proceed until all of it is used up in the reaction
- reacts with melanin to bleach them
ascorbic acid
lower the pH of the fruit tissue to retard the action of phenolase
Citric and acetic acid
pH where phenolase will be deactivated
3.0
restricts the amount of oxygen in contact with apple tissue
soaking in plain watter
cannot be used as treatment for fruits which will be served fresh
heat
TT1 Potato Oxidase (Phenol)
During Exp: Cloudy
Next Lab Period: Red/brown
TT2 Potato Oxidase ( Pyrogallol)
During Exp: colorless
Next lab period: From colorless to purple because it absorbed oxygen from air
toxic white with slightly pink shade crystalline solid
phenol
phenol is also known as
carbolic acid
formula of phenol
C6H6O
white crystalline powder
powerful reducing agen
Pyrogallol
formula of pyrogallol
C6H6O3
uses of pyrogallol
hair drying
developing agent in black and white photos
an enzyme that catalyzes the reduction of H2O2 liberating O and H2O
catakase
Reagents in TT1 Vegetable catalase
potato extract
H2O2
Reagents in TT2 Vegetable Catalase
boiled potato extract
H2O2
Reagents in TT3 (control) vegetable catalase
Water
H2O2
Reaction in TT1 Vegetable catalase
produced bubbles
Reaction in TT2 Vegetable catalase
did not produce bubbles
Reaction in TT3 Vegetable catalase
does not produce bubbles
Factors that influence enzyme activity
concentration of enzyme concentration of substrate pH Inorganic salts of heavy metals Presence of activators (co-enzymes) and inhibitors (anti-enzymes)
Tests for Carbohydrates
Iodine Test Fehling's Test Benedict's Test Barfoed's Test Picric acid Test Seliwanoff's Test Moore's Test Molisch's Test Trommer's Test Mucic Acid Test
- specific test for starch
- test for dextrin
Iodine test
Reagent composition and color of Iodine test
Iodine
brown
Positive result in Iodine Test
Starch - dark blue
Dextrin - purple
- test for sugar
- tests the presence of aldehydes but not ketones
Fehling’s test
Reagent composition and color of Fehling’s test
A - CuSO4
B- NaOH + Na K Tartrate
Dark blue
Positive result of Fehling’s test
Brick red precipitate
- sensitive test for the presence of sugar in urine
- test determine whether a monosaccharide or disaccharide is a reducing sugar
Benedict’s test
Reagent composition and color of Benedict’s test
CuSO4 + Na Citrate + Na2CO3
Light blue
Positive result of Benedict’s test
Brick red precipitate
- differentiates mono and disaccharides
- similar to Fehling’s test, except that different types of sugars react at different rates
Barfoed’s test
Reagent composition and color of Barfoed’s Test
Copper acetate (CuAc) Acetic acid (HAc)
light blue
Positive result of Barfoed’s test
Brick red precipitate
- test for the formation of sodium picramic acid/ picramate
Picric acid test
Reagent composition and color of Picric acid test
Picric acid
yellow
Positive result of Picric acid test
Mahogany red solution
- distinguishes aldose (-) from ketose (-)
- specific test for fructose
Seliwanoff’s test
Reagent composition and color of Seliwanoff’s test
Resorcinol + HCl
yellow
Positive result of Seliwanoff’s test
Cherry red compound/complex
test based on the liberation of aldehyde
Moore’s test
Reagent composition and color of Moore’s test
NaOH
Colorless
Positive result of Moore’s test
Brown color with caramel-like odor
general test for carbohydrates
Molisch’s test
Reagent composition and color of Molisch’s test
alphanaphthol in alcohol layered with H2SO4
red orange
Positive result of Molisch’s test
violet ring
specific test for glucose
Trommer’s test
Reagent composition and color of Trommer’s test
CuSO4 + NaOH
blue
Positive result of Trommer’s test
glucose - red precipitate
water - black
specific test for galactose
Mucic Acid test
Reagent composition and color of Mucic Acid test
HNO3
colorless
Positive result of Mucic Acid test
galactose - white flaked crystals
lactose - glucose interferes making fine elongated crystals
Positive and negative results in Fehling’s test
Positive: glucose, maltose, fructose, lactose, galactose
negative: sucrose
Positive and negative results in Benedict’s test
Positive: glucose, maltose, fructose, lactose, galactose
negative: sucrose
Positive and negative results in Barfoed’s test
Positive: glucose, fructose, galactose
negative: sucrose, maltose, lactose
Positive and negative results in Picric Acid test
Positive: glucose, maltose, fructose, lactose, galactose
negative: sucrose
Positive and negative results in Seliwanoff’s test
Positive: sucrose, fructose
Negative: Glucose, Maltose, galactose, lactose
Positive and negative results in Moore’s test
Positive: glucose, maltose, fructose, lactose, galactose
negative: sucrose
Positive and negative results in Molisch’s test
all positive
- an intermediate product before the final change of starch into sugar
- has same chemical formula as starch but possesses different properties
- smaller and less complex structure to starch that is formed during hydrolysis of starch
Dextrin
formula of dextrin
(C6H10O5)n
- are polyhydroxyaldehydes or polyhydroxy ketones
- substances that yield these compounds upon hydrolysis
carbohydrates
means “containing several alcohol groups”
Polyhydroxy
are alcohols and are also either aldehydes or ketones
simple carbohydrates
Difference of Reducing sugars from non-reducing sugars
Reducing:
- possess a free aldehyde or ketone
- can reduce cupric ions to cuprous ions in Fehling’s or Benedict’s solution that precipitate out as Cu2O cuprous oxide
- oxidized by mild oxidizing agents
Difference of Non reducing sugars from reducing sugars
Non-reducing:
- free aldehyde or ketone is lacking
- no brick red precipitate
- not oxidized by mild oxidizing agents
exist in solution as an equilibrium of open chain and closed ring structures
sugars
carbon atom that contains the C=O bond in the open chain form
carbonyl carbon
the carbonyl carbon is the one which is attached to the O of the ring and an OH group
closed ring (cyclic)
type of sugar that is a reducing sugar
all common monosaccharides
reducing disaccharides
lactose and maltose
disaccharide that is a non-reducing sugar
sucrose
common oxidizing agents used to test for the presence of a reducing sugar
Benedict’s solution
Fehling’s solution
result when closed ring opens to form a chain
alkanal
alkanone
sugars that are aldehydes
aldoses
sugars that are ketones
ketoses
Advantages of Benedict’s solution over Fehling’s
- Sensitive test for glucose thus used in urine test
- More sensitive to reducing sugars
- Not affected by uric acid, creatinine, chloroform, formalin which are sometimes employed as preservatives in urine