Production and Utilization of Enzyme (Quiz 1)

Question 1

Among various enzymes produced at large scale are
_______ (subtilisin, rennet),
_______ (pectinase, lipase, lactase),
_______ (glucose isomerase), and
_______ (glucose oxidase).

These enzymes are produced using overproducing strains of certain organisms.

Answer 1

• proteases
• hydrolases
• isomerases
• oxidases

Question 2

Separation and Purification of an enzyme from an organism require:

Answer 2

• disruption of cells,
• removal of cell debris and nucleic acids,
• precipitation of proteins,
• ultrafiltration of the desired enzyme,
• chromatographic separations (optional),
• crystallization
• drying.

Question 3

The process scheme varies depending on whether the enzyme is ______ or _________.

Answer 3

Intracellular or extracellular

Question 4

In some cases, it may be more advantageous to use __________ cells with the desired enzyme activity in immobilized form. This approach eliminates costly enzyme separation and purification steps and is therefore economically more feasible.

Answer 4

Inactive (dead or resting) Cells

Question 5

The first step in the large-scale production of enzymes is to __________ the organisms producing the desired enzyme.

Answer 5

cultivate

Question 6

________ can be regulated and _________ conditions can be optimized for overproduction of the enzyme

Answer 6

Enzyme production
fermentation

Question 7

__________ are produced by using overproducing strains of Bacillus, Aspergillus, Rhizopus, and Mucor

Answer 7

Proteases

Question 8

__________ are produced by Aspergillus niger

Answer 8

Pectinases

Question 9

________ are produced by yeast and Aspergillus

Answer 9

lactases

Question 10

__________ are produced by certain strains of yeasts and fungi

Answer 10

lipases

Question 11

____________ is produced by Flavobacterium arborescens or Bacillus coagulans.

Answer 11

glucose isomerase

Question 12

Glucose isomerase is produced by __________ or ___________.

Answer 12

Flavobacterium arborescens or Bacillus coagulans

Question 13

After the cultivation step, cells are separated from the media usually by ___________ or sometimes by _________. Depending on the intracellular or extracellular nature of the enzyme

Answer 13

filtration
centrifugation

Question 14

The recovery of intracellular enzymes is more complicated and involves the _______ of cells and removal of cell debris and nucleic acids

Answer 14

disruption

Question 15

In some cases, enzyme may be both intracellular and extracellular, which requires processing of both _____ and ______.

Answer 15

broth and cells.

Question 16

• Intracellular enzymes may be released by increasing the _________ of cell membrane.

Certain salts such as CaCl₂ and other chemicals such as dimethylsulfoxide (DMSO) and pH shift may be used for this purpose.

• If enzyme release is not complete, then cell ________ may be essential.

Answer 16

permeability
cell disruption

Question 17

__________ hydrolyze proteins into smaller peptide units and constitute a large and industrially
important group of enzymes

Answer 17

Proteases

Question 18

_________constitute about 60% of the total enzyme market.

Industrial Proteases are obtained from
- ________ (Bacillus),
- ________ (Aspergillus, Rhizopus, and Mucor),
- ________
- ________

Answer 18

Proteases
- bacteria
- Molds
- Animal pancreas
- plants

Question 19

Most of the industrial proteases are __________

Answer 19

endoproteases

Question 20

Proteases are used in:

• ____________ (rennet)
• ____________
• ____________ (papain, trypsin),
• ____________ (trypsin, pepsin)
• ____________(subtilisin Carlsberg)
• ____________

Answer 20

• food processing, such as cheese making (rennet)
• baking
• meat tenderization (papain, trypsin),
• brewing (trypsin, pepsin)
• detergents for the hydrolysis of protein stains
• tanning and the medical treatment of wounds

Question 21

__________ are produced mainly by A. niger (Aspergillus niger)

Answer 21

Pectinases

Question 22

The major components in pectinases are:

Answer 22

• pectin esterase
• polygalacturonase
• polymethyl galacturonate lyase

Question 23

___________ are used in fruit juice processing and wine making to Increase juice yield, reduce viscosity, and clear the juice.

Answer 23

Pectinases

Question 24

_______ hydrolyze lipids into fatty acids and glycerol and are produced from animal pancreas, some molds, and yeasts.

o may be used to hydrolyze oils for soap manufacture and to hydrolyze the lipid-fat. compounds present in waste-water streams.

Answer 24

Lipases

Question 25

_____________ of **oils and fats may be catalyzed by lipases**. Lipases may also be used in the **cheese and butter industry to impart flavor** as a result of the hydrolysis of fats. o Lipase containing detergents are an important application of lipase

Answer 25

Interesterification

Question 26

_____________ are used for the **hydrolysis of starch** and are **produced by** many different organisms, including **A. niger** (Aspergillus niger) and **B. subtilis** (Bacillus subtilis)

Answer 26

Amylases

Question 27

Three **major types of amylases** are:

Answer 27

- a-amylase - b-amylase - glucoamylase

Question 28

**breaks a -1,4 glycosidic bonds randomly** on the amylose chain and **solubilizes amylose**. For this reason, __________ is known as the __________ enzyme

Answer 28

***a -amylase*** starch- liquefying

Question 29

**________ hydrolyzes a -1,4 glycosidic bonds** on the **nonreducing ends of amylose** and **produces maltose residues**. And is known as a _______

Answer 29

**b-amylase** saccharifying enzyme.

Question 30

a **-1,6 glycosidic linkages in the amylopectin** fraction of starch are **hydrolyzed** by _________, which is **also known as a saccharifying enzyme**.

Answer 30

glucoamylase

Question 31

In the United States on the average, nearly **1.3x109 lb/yr of glucose is produced** by the *enzymatic hydrolysis of starch*. The enzyme _______ also **hydrolyzes a -1,6 glycosidic linkages** in starch **selectively**

Answer 31

pullulanase

Question 32

__________ are used in the **hydrolysis of cellulose** and are produced by some *Trichoderma species*, some *molds*, and by some *Clostridium species* - is an **enzyme complex**, and its formation is induced by cellulose - **Tricho derma cellulase hydrolyzes crystalline cellulose**, but *Aspergillus cellulase* does not

Answer 32

Cellulases

Question 33

**Cellulose** is *first hydrolyzed* to **cellobiose** by cellulase, and __________ is **further hydrolyzed** to _______ by b-glycosidase. Both of these enzymes are Inhibited by their end products, _____and______. (same 2 ans.) * **Cellulases** are used in *cereal processing*, *alcohol fermentation from biomass*, *brewing*, and *waste treatment*

Answer 33

cellobiose and glucose

Question 34

___________ **hydrolyze** hemicellulose to **five-carbon sugar units** and are produced by some **molds**, such as white rot fungi and A. niger - are used in **combination with other enzymes** in baking doughs, brewing mashes, alcohol fermentation from biomass, and waste treatment.

Answer 34

Hemicellulases

Question 35

_______ are used to **hydrolyze lactose in whey to glucose** and **galactose** and are produced by *yeast* and some *Aspergillus species*. o are used in the fermentation of cheese whey to ethanol

Answer 35

Lactases

Question 36

Other microbial ________ produced by **Bacillus amyloliquefaciens**, **A. niger**, and **Penicillium emersonii** are used in *brewing mashes* containing barley or malt. These enzymes **improve wort filtration** and **extract yield**.

Answer 36

glucanases

Question 37

_____________ is used by the **antibiotic industry** to convert *penicillin G to 6-aminopenicillanic acid (6-APA)*, which is a **precursor for semisynthetic penicillin derivatives**

Answer 37

Penicillin acylase

Question 38

**For the conversion of fumarate to L-aspartate by aspartase**, - in industry, this conversion is realized in a _____ of immobilized **dead E. coli cells with active aspartase** enzyme. - ______ solution is passed through the column, and **aspartate is obtained in the ____ stream**. - **Aspartate** is further **coupled with L-phenylalanine** to produce ______, which is a **low-calorie sweetener** known as "___________."

Answer 38

* **packed column** * **Fumarate** solution, * **effluent** * **aspartame**, * **Nutrasweet**

Question 39

______ is nearly **1.7 times sweeter than glucose** and is used as a **sweetener in soft drinks**. Glucose isomerase is an intracellular enzyme

Answer 39

Fructose

Question 40

Some **Industrially Important Enzymes** pt. 1 1. Application: **Starch hydrolysis**, **glucose production** Ex. of Source: *Bacillus subtilis*, *Aspergillus Niger* 2. Application: **Saccharification of starch**, **glucose production** Ex. of Source.: *A. niger*, *Rhizopus niveus*, *endomycopsis* 3. Application: **Meat tenderizer**, **beer haze removal** Ex. of Source: *Animal Pancreas* 4. Application: **Digestive aid**, **meat tenderizer**, **Medical applications** Ex. of Source: *Papaya*

Answer 40

1. Amylase 2. Glucoamylase 3. Trypsin 4. Papain

Question 41

Some **Industrially Important Enzymes** pt. 2 5. Application: **Digestive aid**, **meat tenderizer** Ex. of Source: *Animal stomach* 6. Application: **Cheese manufacturing** Ex. of Source: *Calf stomach/recombinant E. coli* 7. Application: **Isomerization of glucose to fructose** Ex. of Source: *Flavobacterium arborescens*, *Bacillus cougulans*, *Lactobacillus brevis* 8. Application: **Degradation of penicillin** Ex. of Source: *B. subtilis* 9. Application: **Glucose to gluconic acid**, **dried-egg manufacture** Ex. of Source: *A. niger*

Answer 41

5. Pepsin 6. Rennet 7. Glucose isomerase 8. Penicillinase 9. Glucose oxidase

Question 42

Some **Industrially Important Enzymes** pt. 3 10. Application: **Biopulping of wood for paper manuf.** Ex. of Source: *Fungal* 11. Application: **Hydrolysis of lipids**, **Flavoring and digestive aid** Ex. of Source: *Rhizopus*, *pancreas* 12. Application: **Hydrolysis of sucrose** for further fermentation Ex. of Source: *S. cerevisae* 13. Application: **Clarification of fruit juice**, **hydrolysis of pectin** Ex. of Source: *A. oryzae*, *A. niger*, *A. flavus* 14. Application: **cellulose hydrolysis** Ex. of Source: *Trichoderma viride*

Answer 42

10. Lignases 11. Lipases 12. Invertase 13. Pectinase 14. Cellulase

Question 43

___________ are commonly used in **medicine for diagnosis**, **therapy**, and **treatment** purposes.

Answer 43

Enzymes

Question 44

1. __________ **can be** used as an **anti-inflammatory agent** 2. _________ **is used** as an **anti-inflammatory agent**

Answer 44

1. Trypsin 2. streptokinase

Question 45

_______, which **hydrolyzes the cell wall of gram-positive bacteria**, is used as an **antibacterial agent**;

Answer 45

Lysozyme

Question 46

_______ is used in **dissolving and preventing blood clots**.

Answer 46

urokinase

Question 47

__________, which catalyzes the conversion of L-asparagine to L- aspartate, is used as an **anticancer agent**. And is produced by E. Coli

Answer 47

Asparaginase

Question 48

___________ catalyzes the **oxidation of glucose to gluconic acid and hydrogen peroxide**, which can easily be detected. And is used for the **determination of glucose levels in blood and urine**.

Answer 48

Glucose oxidase

Question 49

_________ **hydrolyze penicillin** and are used to **treat allergic reactions against penicillin**.

Answer 49

Penicillinases

Question 50

__________ (TPA) and ______ are used in the **dissolution of blood clots** (particularly following a **heart attack** or **stroke**).

Answer 50

Tissue plasminogen activator and streptokinase

Question 51

The **development of biosensors using enzymes** as integral components is proceeding rapidly. **Two examples of immobilized enzyme electrodes** are those used in the **determination of glucose and urea** by using _______ and ________ immobilized on the **electrode membrane**, respectively

Answer 51

glucose oxidase and urease