1. Industrial use of enzymes

2. The large-scale use of enzymes in solution
 starch processing,
high-fructose syrup manufacture,
textile desizing and
detergent formulation

3. The use of enzymes in detergents
The use of enzymes in detergent formulations is now common in developed countries, with over half of all detergents presently available containing enzymes.
Dirt comes in many forms and includes proteins, starches and lipids. In addition, clothes that have been starched must be freed of the starch. Using detergents in water at high temperatures and with vigorous mixing, it is possible to remove most types of dirt but the cost of heating the water is high and lengthy mixing or beating will shorten the life of clothing and other materials. The use of enzymes allows lower temperatures to be employed and shorter periods of agitation are needed, often after a preliminary period of soaking.

4. Detergent enzymes must be cost-effective and safe to use. 
The enzymes used are all produced using species of Bacillus, mainly by just two companies. Novo Industri A/S produce and supply three proteases, Alcalase, from B. licheniformis, Esperase, from an alkalophilic strain of a B. licheniformis and Savinase, from an alkalophilic strain of B. amyloliquefaciens (often mistakenly attributed to B. subtilis).

5. Compositions of an enzyme detergent

6. Applications of proteases in the food industry
Certain proteases have been used in food processing for centuries and any record of the discovery of their activity has been lost in the mists of time.
Rennet (mainly chymosin), obtained from the fourth stomach (abomasum) of unweaned calves has been used traditionally in the production of cheese. 

7. Similarly, papain from the leaves and unripe fruit of the pawpaw (Carica papaya) has been used to tenderise meats.
These ancient discoveries have led to the development of various food applications for a wide range of available proteases from many sources, usually microbial.

8. When proteases are used to depolymerise proteins, usually non-specifically, the extent of hydrolysis (degree of hydrolysis) is described in DH units where:

10. Baking industry
Meat tenderisation
Leather industry
Wool industry

11. Enzymes for starch conversion
a-amylase randomly hydrolyse a-1,4 linkages in both amylose and amylopectin to yeild mixture of glucose, maltose, maltotriose and series of a-limit dextrins.
b-amylase sometimes used in place of a-amylase. They hydrolyze alternate a-1,4 linkages and yield maltose residues and b-limit dextrins
Glucoamylase hydrolyses a-1,3. a-1,4 and a-1,6 linkages but is less efficient than a-amylase. Major role is to break cross links of amylopectin resulting in complete breakdown to glucose. Generally used to reduce CHO content of beers. Industrially obtained from fungus Aspergillus niger.
Glucose isomerase is used for conversion of glucose obtained after processing to fructose.
Pullulanase (pullulan a-1,6-glucanohydrolase) or isoamy- lase (glycogen a-1,6-glucanohydrolase) cleaves the a-1,6- linked branch points of starch and produces linear amylosaccharides of varying lengths.

12. Production of glucose syrup Production of high fructose corn syrup
Production of high maltose conversion syrups
Production of D-glucose from starch by acid hydrolysis (chemical) produces undesirable bitter sugar (gentiobiose), and the inevitable formation of salt (from subsequent neutralization with alkali) and coloring materials.
With the discovery and development of thermostable a-amylase from Bacillus licheniformis, an enzymatic process has replaced the acid hydrolysis process.
liquefaction and saccharification
Typically, glucose syrups (DE 97–98) having 96% glucose contain 2–3% disaccharides (maltose and isomaltose) and 1–2% higher saccharides.

13. The use of enzymes in starch hydrolysis, HFCS
There are three stages in the conversion of starch
gelatinisation, involving the dissolution of the nanogram-sized starch granules to form a viscous suspension; 
liquefaction, involving the partial hydrolysis of the starch, with concomitant loss in viscosity; and
saccharification, involving the production of glucose and maltose by further hydrolysis.

14. The starch and glucose syrup industry uses the expression dextrose equivalent or DE, similar in definition to the DH units of proteolysis, to describe its products, where:

16. 3. Various maltose-containing syrups are used in the brewing, baking, soft drink, canning, confectionery, and other food industries.
There are three types of maltose-containing syrups:
high-maltose syrup (DE 35–50, 45–60% maltose, 10–25% maltotriose, 0.5–3% glucose),
extra high-maltose syrup (DE 45–60, 70–85% maltose, 8–21% maltotriose, 1.5–2% glucose), and
high conversion syrup (DE 60–70, 30–47% maltose, 35–43% glucose, 8–15% maltotriose).
liquefaction and saccharification, as in the production of glucose.
However, in this process, the liquefaction reaction is terminated when the DE reaches about 5–10 since a low DE value increases the potential for attaining high maltose content.
maltogenic amylase such as b-amylase, b-amylase with pullulanase or isoamylase, or a fungal a-amylase at pH 5.0–5.5 and 50–55o C.

17. Enzymmes used in starch hydrolysis

18. Application of Carbohydrases
Cellulase - hydrolyzes cellulose
Cellulose is a polysaccharide (glucose units)
Stonewashing genes
break down fabric (which is cellulose)
releases dye

19. Cellulose (C6H10O5)n
Structural component of cell wall of green plants, many algae and fungi. Some bacteria secrete it to form biofilms.
It is the most common organic compound on earth.
33% plant matter is cellulose. Cotton is 90% cellulose and wood is 50% cellulose.
Industrially, cellulose is obtained from wood pulp and cotton to produce cardboard and paper and derivatized to make cellophane and rayon.
Cellulose can be digested in the gut of ruminants and termites with the help of symbiotic bacteria (Trichonympha, which produces cellulases). Humans cannot digest cellulose but acts as dietary fiber and hydrophilic bulking agent for faces.
The major combustible component of non-food energy crops is cellulose, with lignin being second.
Some bacteria can convert cellulose into ethanol which can then be used as a fuel .
Cellulose is crystalline, strong, and resistant to hydrolysis, cellulose contains only anhydrous glucose residues with beta configuration.

20. Structural Unit
Cellulose is a linear polymer of β-(1,4)-D-glucopyranose units in 4C1 conformation. The fully equatorial conformation of β-linked glucopyranose residues stabilizes the chair structure, minimizing its flexibility (for example, relative to the slightly more flexible α-linked glucopyranose residues in amylose).

21. Functionality
Cellulose has many uses as an anticake agent, emulsifier, stabilizer, dispersing agent, thickener, and gelling agent but these are generally subsidiary to its most important use of water holding capacity.
Water cannot penetrate crystalline cellulose but dry amorphous cellulose absorbs water becoming soft and flexible. Some of this water is non-freezing but most is simply trapped.
Less water is bound by direct hydrogen bonding if the cellulose has high crystallinity but some fibrous cellulose products can hold on to considerable water in pores and its typically straw-like cavities; water holding ability correlating well with the amorphous (surface area effect) and void fraction (that is, the porosity).
As such water is supercoolable, this effect may protect against ice damage. Cellulose can give improved volume and texture particularly as a fat replacer in sauces and dressings but its insolubility means that all products will be cloudy.

22. Lignocellulosics Primary cellulosics Agricultural waste cellulosics
Lignocellulose: structural support system for all terrestrial plants
Plant biomass comprises of Lignin, hemicellulose and cellulose combined in different proportions
Lignocellulosics
Primary
cellulosics
Agricultural
waste
cellulosics
Municipal
Waste
cellulosics
Plant material that remain after harvesting and processing Straw, corn, rice hulls, sugarcane baggase, animal manure, timber residues
Plants harvested for cellulosic content, structural use or feed
Cotton, timber, hay
Waste paper and discarded paper products

23. Enzymes for cellulose hydrolysis
Fungus: Trichoderma reesei
Cellulomonas fimi
Aspergillus
Endo-1,4-b-glucanse: hydrolyzes b-1,4 linkages b/w adj glu mocs (cellulase, EC )
Exo-1,4-b-glucanase: degrades nicked cellulose chains from non reducing ends and produced glucose, cellobiose (2 glu units) and cellotriose (3 glu units)
1,4-b-Cellobiohydrolase; type of exoglucanase removes units of 10 or more glu residues from non reducing ends (found in cellulolytic fungi)
b-glucosidase or cellobiase converts cellobiose and cellotriose to glucose
cellobiose

24. Enzymatic biodegradation of cellulose
Crystalline region
Amorphous region
Endoglucanase
Enzymatic biodegradation of cellulose
Removal of oligosacc. from reducing ends
Exoglucanase
glu
cellobiose
Endoglucanase
cellotriose
Exoglucanase
b-glucosidase
Cellobiohydrolase

25. b-glucosidase enhances enzymatic utilization of cellulose
Cellobiose
Glucose
Inhibits
Feedback Inhibitor of cellobiose
Feedback Inhibitor of cellulose hydrolysis
Fermentation
b-glucosidase not only produces glucose from cellobiose but also lowers cellobiose inhibition, allowing the cellulolytic enzymes to function more efficiently.
However, like b-glucanases, most b-glucosidases are subject to product (glucose) inhibition.
Decrease amount of cellobiose which prevents end product inhibition of exo and endo
b-glucosidase
Cloning of gene in host cell
To increase rate and extent of degradation , addition of
b-glucosidase enhances enzymatic utilization of cellulose

26. Enzymes use in Molecular biology
1. Restriction endonucleases
defense
specific sequence
methylation
blunt vs staggered
4, 6, 8 base cutters
2. Ligase catalyze formation of bonds of nucleic acids (DNA)
3. DNA polymerase
taq
Deep Vent DNA polymerase