Lesson 4.4. Hydrolysis of Polysaccharides

5-mL of 1% Starch solution

ACID HYDROLYSIS ENZYMATIC HYDROLYSIS

1.Transfer _______________________ (Distilled water) in a clean hard test tube.

2.Add __________________ and mix thoroughly.

3.Cover the test tube with a ________________________.

4.Digest the contents of the sealed test tube in a boiling water bath for _______.

5.After digestion, remove the test tube from the water bath and allow it to ______.

6.Neutralize the hydrolysate by adding ____________________ until the solution reaches a neutral pH.

7.Keep the neutral hydrolyzed product for ____________________.

1 = ?

2.5 mL of 6M HCl

ACID HYDROLYSIS ENZYMATIC HYDROLYSIS

1.Transfer _______________________ (Distilled water) in a clean hard test tube.

2.Add __________________ and mix thoroughly.

3.Cover the test tube with a ________________________.

4.Digest the contents of the sealed test tube in a boiling water bath for _______.

5.After digestion, remove the test tube from the water bath and allow it to ______.

6.Neutralize the hydrolysate by adding ____________________ until the solution reaches a neutral pH.

7.Keep the neutral hydrolyzed product for ____________________.

2 = ?

cork and seal with a parafilm

ACID HYDROLYSIS ENZYMATIC HYDROLYSIS

1.Transfer _______________________ (Distilled water) in a clean hard test tube.

2.Add __________________ and mix thoroughly.

3.Cover the test tube with a ________________________.

4.Digest the contents of the sealed test tube in a boiling water bath for _______.

5.After digestion, remove the test tube from the water bath and allow it to ______.

6.Neutralize the hydrolysate by adding ____________________ until the solution reaches a neutral pH.

7.Keep the neutral hydrolyzed product for ____________________.

3 = ?

1 hour

ACID HYDROLYSIS ENZYMATIC HYDROLYSIS

1.Transfer _______________________ (Distilled water) in a clean hard test tube.

2.Add __________________ and mix thoroughly.

3.Cover the test tube with a ________________________.

4.Digest the contents of the sealed test tube in a boiling water bath for _______.

5.After digestion, remove the test tube from the water bath and allow it to ______.

6.Neutralize the hydrolysate by adding ____________________ until the solution reaches a neutral pH.

7.Keep the neutral hydrolyzed product for ____________________.

4 = ?

cool

ACID HYDROLYSIS ENZYMATIC HYDROLYSIS

1.Transfer _______________________ (Distilled water) in a clean hard test tube.

2.Add __________________ and mix thoroughly.

3.Cover the test tube with a ________________________.

4.Digest the contents of the sealed test tube in a boiling water bath for _______.

5.After digestion, remove the test tube from the water bath and allow it to ______.

6.Neutralize the hydrolysate by adding ____________________ until the solution reaches a neutral pH.

7.Keep the neutral hydrolyzed product for ____________________.

5 = ?

6M NaOH dropwise

ACID HYDROLYSIS ENZYMATIC HYDROLYSIS

1.Transfer _______________________ (Distilled water) in a clean hard test tube.

2.Add __________________ and mix thoroughly.

3.Cover the test tube with a ________________________.

4.Digest the contents of the sealed test tube in a boiling water bath for _______.

5.After digestion, remove the test tube from the water bath and allow it to ______.

6.Neutralize the hydrolysate by adding ____________________ until the solution reaches a neutral pH.

7.Keep the neutral hydrolyzed product for ____________________.

6 = ?

further analysis

ACID HYDROLYSIS ENZYMATIC HYDROLYSIS

1.Transfer _______________________ (Distilled water) in a clean hard test tube.

2.Add __________________ and mix thoroughly.

3.Cover the test tube with a ________________________.

4.Digest the contents of the sealed test tube in a boiling water bath for _______.

5.After digestion, remove the test tube from the water bath and allow it to ______.

6.Neutralize the hydrolysate by adding ____________________ until the solution reaches a neutral pH.

7.Keep the neutral hydrolyzed product for ____________________.

7 = ?

1% Starch solution

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

1 = ?

1 mL of amylase enzyme

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

2 = ?

3x with clean water

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

2.a. = ?

2 ml of saliva

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

2.b. = ?

5 ml PB pH 7.0

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

2.c. = ?

1 mL of 5% acarbose

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

3 = ?

Cap

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

4 = ?

40*C oven for 1 hour

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

5 = ?

boiling water bath

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

6 = ?

further analysis

ENZYMATIC HYDROLYSIS:

1.Prepare two 5-mL of ______________ (use PBS buffer) in a separate 10-mL centrifuge tube and label with CT1 and CT2.

2.Add ___________________ into CT1 and CT2 and mix thoroughly.

a. Collection of amylase (saliva): Rinse the mouth ___________________

b. Spit at least ___________________ in a beaker

c. Add ___________________

3.Add ___________________ into CT1 (this will serve as the inhibited tube)

4._____ tightly the centrifuge tube

5.Incubate the test tube in a ______________________.

6.After incubation, heat the hydrolysate in a ____________ to denature the enzyme.

7.Keep the enzyme hydrolyzed product for ______________.

7 = ?

Acid Hydrolysis

complete hydrolysis

monosaccharide product

Acid hydrolysis yields:

Enzymatic hydrolysis

incomplete hydrolysis that breaks specific bonds

mixture of glucose, maltose & other hydrolysates

In enzymatic hydrolysis, the salivary amylase will break down α-1,4 glycosidic bonds and yield a _________________________

Basic Hydrolysis

• aldo/keto groups are sensitive to strong base attacks

• epimerization of aldoses & ketoses and aldose-ketose interconversion