Production and Utilization of Enzyme (Quiz 1) Flashcards
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.
- proteases
- hydrolases
- isomerases
- oxidases
Separation and Purification of an enzyme from an organism require:
- disruption of cells,
- removal of cell debris and nucleic acids,
- precipitation of proteins,
- ultrafiltration of the desired enzyme,
- chromatographic separations (optional),
- crystallization
- drying.
The process scheme varies depending on whether the enzyme is ______ or _________.
Intracellular or extracellular
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.
Inactive (dead or resting) Cells
The first step in the large-scale production of enzymes is to __________ the organisms producing the desired enzyme.
cultivate
________ can be regulated and _________ conditions can be optimized for overproduction of the enzyme
Enzyme production
fermentation
__________ are produced by using overproducing strains of Bacillus, Aspergillus, Rhizopus, and Mucor
Proteases
__________ are produced by Aspergillus niger
Pectinases
________ are produced by yeast and Aspergillus
lactases
__________ are produced by certain strains of yeasts and fungi
lipases
____________ is produced by Flavobacterium arborescens or Bacillus coagulans.
glucose isomerase
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
filtration
centrifugation
The recovery of intracellular enzymes is more complicated and involves the _______ of cells and removal of cell debris and nucleic acids
disruption
In some cases, enzyme may be both intracellular and extracellular, which requires processing of both _____ and ______.
broth and cells.
- 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.
permeability
cell disruption
__________ hydrolyze proteins into smaller peptide units and constitute a large and industrially
important group of enzymes
Proteases
Proteases constitute about 60% of the total enzyme market.
Industrial Proteases are obtained from
- ________ (Bacillus),
- ________ (Aspergillus, Rhizopus, and Mucor),
- ________
- ________
- bacteria
- Molds
- Animal pancreas
- plants
Most of the industrial proteases are __________
endoproteases
Proteases are used in:
- ____________ (rennet)
- ____________
- ____________ (papain, trypsin),
- ____________ (trypsin, pepsin)
- ____________(subtilisin Carlsberg)
- ____________
- 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
__________ are produced mainly by A. niger (Aspergillus niger)
Pectinases
The major components in pectinases are:
- pectin esterase
- polygalacturonase
- polymethyl galacturonate lyase
___________ are used in fruit juice processing and wine making to Increase juice yield, reduce viscosity, and clear the juice.
Pectinases
_______ 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.
Lipases
_____________ 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
Interesterification
_____________ are used for the hydrolysis of starch and are produced by many different organisms, including A. niger (Aspergillus niger) and B. subtilis (Bacillus subtilis)
Amylases
Three major types of amylases are:
- a-amylase
- b-amylase
- glucoamylase
breaks a -1,4 glycosidic bonds randomly on the amylose chain and solubilizes amylose.
For this reason, __________ is known as the __________ enzyme
a -amylase
starch- liquefying
________ hydrolyzes a -1,4 glycosidic bonds on the nonreducing ends of amylose and produces maltose residues.
And is known as a _______
b-amylase
saccharifying enzyme.
a -1,6 glycosidic linkages in the amylopectin fraction of starch are hydrolyzed by _________,
which is also known as a saccharifying enzyme.
glucoamylase
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
pullulanase
__________ 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
Cellulases
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
cellobiose and glucose
___________ 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.
Hemicellulases
_______ 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
Lactases
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.
glucanases
_____________ is used by the antibiotic industry to convert penicillin G to 6-aminopenicillanic
acid (6-APA), which is a precursor for semisynthetic penicillin derivatives
Penicillin acylase
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 “___________.”
- packed column
- Fumarate solution,
- effluent
- aspartame,
- Nutrasweet
______ is nearly 1.7 times sweeter than glucose and is used as a sweetener in soft drinks. Glucose isomerase is an intracellular enzyme
Fructose
Some Industrially Important Enzymes pt. 1
- Application: Starch hydrolysis, glucose production
Ex. of Source: Bacillus subtilis, Aspergillus Niger - Application: Saccharification of starch, glucose production
Ex. of Source.: A. niger, Rhizopus niveus, endomycopsis - Application: Meat tenderizer, beer haze removal
Ex. of Source: Animal Pancreas - Application: Digestive aid, meat tenderizer, Medical applications
Ex. of Source: Papaya
- Amylase
- Glucoamylase
- Trypsin
- Papain
Some Industrially Important Enzymes pt. 2
- Application: Digestive aid, meat tenderizer
Ex. of Source: Animal stomach - Application: Cheese manufacturing
Ex. of Source: Calf stomach/recombinant E. coli - Application: Isomerization of glucose to fructose
Ex. of Source: Flavobacterium arborescens, Bacillus cougulans, Lactobacillus brevis - Application: Degradation of penicillin
Ex. of Source: B. subtilis - Application: Glucose to gluconic acid, dried-egg manufacture
Ex. of Source: A. niger
- Pepsin
- Rennet
- Glucose isomerase
- Penicillinase
- Glucose oxidase
Some Industrially Important Enzymes pt. 3
- Application: Biopulping of wood for paper manuf.
Ex. of Source: Fungal - Application: Hydrolysis of lipids, Flavoring and digestive aid
Ex. of Source: Rhizopus, pancreas - Application: Hydrolysis of sucrose for further fermentation
Ex. of Source: S. cerevisae - Application: Clarification of fruit juice, hydrolysis of pectin
Ex. of Source: A. oryzae, A. niger, A. flavus - Application: cellulose hydrolysis
Ex. of Source: Trichoderma viride
- Lignases
- Lipases
- Invertase
- Pectinase
- Cellulase
___________ are commonly used in medicine for diagnosis, therapy, and treatment purposes.
Enzymes
- __________ can be used as an anti-inflammatory agent
- _________ is used as an anti-inflammatory agent
- Trypsin
- streptokinase
_______, which hydrolyzes the cell wall of gram-positive bacteria, is used as an antibacterial agent;
Lysozyme
_______ is used in dissolving and preventing blood clots.
urokinase
__________, which catalyzes the conversion of L-asparagine to L- aspartate, is used as an anticancer agent. And is produced by E. Coli
Asparaginase
___________ 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.
Glucose oxidase
_________ hydrolyze penicillin and are used to treat allergic reactions against penicillin.
Penicillinases
__________ (TPA) and ______ are used in the dissolution of blood clots (particularly following a heart attack or stroke).
Tissue plasminogen activator and streptokinase
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
glucose oxidase and urease
