Biotechnology based cleaning agents such as enzyme based agents, are widely used in industries.

The biotech nology based cleaning agents are cheaper and less harmful to the environment. They have specific cleaning action and can also be used at lower temperatures. They produce effluents with lower COD and noncorrosive nature. Enzyme based cleaners are becoming increasingly popular as compared to caustic or acid cleaning regimes. Enzymes used in detergents must be effective at low levels, compatible with various detergent components and to be active at wide range of p! " temperatures. #our classes of enzymes are generally used in detergents i$ %roteases ii$ &mylases iii$ 'ipases iv$ Cellulases

Proteases (ost widely used enzymes in the detergent industry, it removes stains such as grass, blood, egg " human sweat which have tendency to adhere strongly to te)tile fibers. *emoval of proteins by non enzymatic detergents can result in permanent stains due to o)idation and denaturing caused by bleaching and drying. %roteases hydrolyze proteins and brea+ them down into more soluble polypeptides or free amino acids. &s a result of the combined effect of surfactants and enzymes, such hard to remove stains can be removed from fibers. The enzyme protease is produced from al+aliphilic Bacillus clausii ,-( ,./ and strain ,% 01 and Bacillus sp. strain ,-(,%01 and have been incorporated into laundry detergents. -ubtilisin li+e serine proteases belonging to family & of subtilase super family has been used in laundry and dishwashing detergents Amylases 2t is used to remove residues of starch based foods li+e potatoes, spaghetti, custards, gravies and chocolate . &mylase is produced from various organisms. The amylases used in the detergents are produced from bacteria or recombinant organisms. -ome e)amples are Bacillus amyloliquefaciens, al+alophilic Bacillus sp. " thermostable Bacillus licheniformis. Lipases Decompose fatty material. 'ipase is capable of removing fatty stains such as fats, butter, salad oil, sauces and the tough stains on collars and cuffs. 'ipases which are stable and wor+ at al+aline p! 34 to ..$, suitable for enzymated detergent powders " li5uids have good potential for use in detergent industry *ecombinant enzyme Donor Humicola lanuginosa e)pressed in Aspergillus oryzae, *ecombinant enzyme Donor Pseudomonas mendocina e)pressed in Bacillus sp. Cellulases (odify the structure of cellulose fiber on cotton and cotton blends. 6hen it is added to a detergent, it results in7 color brightening, softening and soil removal. & number of al+aliphilic Bacillus produce al+aline cellulose 3carbo)ymethylcellulase$ that is used as an additive for improving the cleaning effect of detergents. 2t is also produced from the fungi Humicola insolens. &mong other enzymes used in some detergents are 8uardzyme 3a pero)idase$ which inhibits dye transfer and Carezyme removing the fuzz that builds up on cotton clothes.

One of the oldest industrial uses of enzyme activity is in leather processing. Before raw hide is transformed into leather, it undergoes a series of operations whereby leather ma+ing protein collagen present in hides and s+ins is freed or partially freed from non collagenous constituents. !ides and s+ins contain fat as well as globular proteins, viz. albumin, globulin, mucoids and fibrous protein such as elastin, +eratin and reticulin between collagen fibers. 2n industry, raw material is processed through a series of operations including soa+ing, liming, dehairing, deliming, bating, degreasing and pic+ling. 2n pre tanning operations, s+ins and hides are sub9ected to a water soa+, which cleans the raw material and loosens the hair. The conventional and most widely used method for dehairing is the treatment of soa+ed raw material with lime and sodium sulphide. -ubse5uent deliming is also done to remove adsorbed lime from the hide. #at present in s+ins is usually removed with degreasing agents such as soluble lime soap, +erosene, chlorinated hydrocarbons or spirit. 2n the traditional processing, a large numbers of pollution causing chemicals, lime, sodium sulphide solvents are released in effluents, which are to)ic and cause environmental pollution. 2n addition of chemical treatment, an enzymatic treatment, +nown, as :bating; is an essential step to obtain optimum results. During bating, the scud is loosened and many unwanted proteins are removed. Bating ma+es the grain surface of the finished leather clean, fine and glossy. The traditional practice of bating is an unhygienic process where uncontrolled fermentation of leather is conducted with manures of dog, pigeon or hen as sources of microorganisms. This process gives a desired character to the finished leather. <o chemical process has been developed which can substitute the fermentation. The leather industry has a ma9or problem regarding industrial pollution due to the use of huge amounts of to)ic chemicals and biological pollution by addition of un+nown microbial load to the environment. =se of enzyme in pre tanning processes appeared to be a viable alternative technology where pollution problems resulting from tannery effluent could be significantly regulated or restricted. Enzymes in pre-tanning %roteolytic enzyme is the most important enzyme used in pre tanning process. Enzymes from plant, animals and microbial sources were +nown to leather industry for a long time. But development of enzyme based process became bright with the success of production of enzymes at commercial level and availability of enzymatic formulations at cheap rate. &nimal proteases and microbial proteases from bacteria and fungi are used in leather industry. The physicochemical properties of the enzyme such as substrate specificity, temperature and p! stability and p! activity range are very important factors in the application of enzymes in different steps of pre tanning operations. (icrobial enzymes appeared to be ideal source of the proteases. Enzymes with wide p! range of activities could be produced economically from different microbes. <ow various commercial enzyme preparations are available for use in leather industry. Enzymes in soaking %roteolytic enzyme combinations 3Aspergillus parasiticus, Aspergillus flavus, Bacillus subtilis, and Aspergillus awamori$ active in natural or al+aline p! ranges are usually used. Enzymes in dehairing =se of enzyme in dehairing is highly desired in leather procesing. 2t would eliminate the use of sodium sulphide, one of the most to)ic chemicals with obno)ious odour. & large number of proteases from A. flavus, A fumigatus, A. chraceus, A. effuses, Bacillus sp., Streptomyces sp. have dehairing or hair loosening effects. %otential use of specific protease +eratinase from Streptomyces fradiae, for dehairing was also indicated. Enzymes from fungal or bacterial sources are allowed to act at p! .>.> for about .? @ ./ hours, and hair is removed by mechanical means. Enzymatic dehairing in tanneries has been envisaged as an alternative to sulfides 3Beynon " Bond, .A4A7 &ltschul et al., .AAB$. &l+aline proteases can be used which enables the swelling of hair roots, and the subse5uent attac+ of protease on the hair follicle protein allowing easy removal of the hair 38upta et al. ?>>?$. The advantages of enzymatic dehairing are as followsC .$ significant reduction or even complete elimination of the use of sodium sulfide, ?$ total recovery of hair resulting good 5uality with good saleable value, and 1$ creation of an ecologically conducive atmosphere for the wor+ers. Enzymes in bating The process of bating is a method for softening hides by treating them in a warm infusion of animal dung. Deliming and proteolytic actions ta+e place simultaneously in bating. & :bate; usually contains a proteolytic enzyme, a carrier

li+e wood flour and deliming agents li+e <! 0Cl or 3<!0$?-O0. %ancreatic enzymes, bacterial and fungal proteases of neutral and al+aline types are used in bating.

Mismatch Repair -ometimes the D<& polymerase enzyme will incorporate the bases which are not complementary to the base present in the template D<& strand. D<& polymerase leaves behind one net error for every .> / to .>4 bases added. Correction of these mista+es done during the replication is accomplished by the mismatch repair. Correction of the rare mismatches left after replication in . coli improves the overall fidelity of replication by an additional factor of .> ? to .>1. The mismatches are nearly always corrected to reflect the information in the old 3template$ strand. -o the repair system must somehow discriminate between the template and the newly synthesized strand. The cell accomplishes this by tagging the template D<& with methyl groups to distinguish it from newly synthesized strands. The mismatch repair system of . coli includes at least .? protein components that function either in strand discrimination or in the repair process itself.

(ut' protein forms a comple) with (utprotein, and the comple) binds to all mismatched base pairs 3e)cept C@C$. (ut! protein binds to (ut' and to 8&TC se5uences encountered by the (ut' (utcomple). D<& on both sides of the mismatch is threaded through the (ut' (ut- comple), creating a D<& loop7 simultaneous movement of both legs of the loop through the comple) is e5uivalent to the comple) moving in both directions at once along the D<&. (ut! has a site specific endonuclease activity that is inactive until the comple) encounters a hemimethylated 8&TC se5uence. &t this site, (ut! catalyzes cleavage of the unmethylated strand on the DE side of the 8 in 8&TC, which mar+s the strand for repair.

6hen the mismatch is on the DE side of the cleavage site, the unmethylated strand is unwound and degraded in the 1EFDE direction from the cleavage site through the mismatch, and this segment is replaced with new D<&. This process re5uires the combined action of D<& helicase 22, --B, e)onuclease 2 or e)onuclease G 3both of which degrade strands of D<& in the 1EFDE direction$, D<& polymerase 222, and D<& ligase. The pathway for repair of mismatches on the 1E side of the cleavage site is similar, e)cept that the e)onuclease is either e)onuclease H22 3which degrades single stranded D<& in the DEF1E or 1EFDE direction$ or *ecI nuclease 3which degrades single stranded D<& in the DEF1E direction$. Base-Excision Repair Base e)cision repair system helps in the removal of wrong bases present in the D<&, which arise due to the spontaneous deamination. #or e)ample when the cytosine undergoes deamination it leads to the formation of the adenine which leads to 8 C to & T mutation. Every cell has a class of enzymes called DNA glycosylases that recognize particularly common D<& lesions 3such as the products of cytosine and adenine deamination$ and remove the affected base by cleaving the ! glycosyl bond. This cleavage creates an apurinic or apyrimidinic site in the D<&, commonly referred to as an AP site or abasic site. Each D<& glycosylase is generally specific for one type of mista+e. #or e)ample, =racil D<& glycosylases, found in most cells, specifically remove the uracil that results from spontaneous deamination of cytosine from D<&. This glycosylase does not remove uracil residues from *<& or thymine residues from D<&.

..

& D<& glycosylase recognizes a damaged base and cleaves between the base and deo)yribose in the bac+bone. &n &% endonuclease cleaves the phosphodiester bac+bone near the &% site. D<& polymerase 2 initiates repair synthesis from the free 1E hydro)yl at the nic+, removing 3with its DEF1E e)onuclease activity$ a portion of the damaged strand and replacing it with undamaged D<&.

?. 1.

0. The nic+ remaining after D<& polymerase 2 has dissociated is sealed by D<& ligase.

Nucleoti e-Excision Repair (ista+es in D<& that affects large portions of the helical structure of D<& are generally repaired by the nucleotide e)cision system. 2n nucleotide e)cision repair, a multisubunit enzyme hydrolyzes two phosphodiester bonds, one on either side of the distortion caused by the mista+e. 2n . coli and other pro+aryotes, the enzyme system hydrolyzes the fifth phosphodiester bond on the 1E side and the eighth phosphodiester bond on the DE side to generate a fragment of .? to .1 nucleotides 3depending on whether the lesion involves one or two bases$. 2n . coli, the +ey enzymatic comple) is the &BC e)cinuclease, which has three subunits, =vr& 3 "r .>0,>>>$, =vrB 3"r B4,>>>$, and =vrC 3"r /4,>>>$. & comple) of the =vr& and =vrB proteins 3&?B$

scans the D<& and binds to the site of a lesion. The =vr& dimer then dissociates, leaving a tight =vrB D<& comple). =vrC protein then binds to =vrB, and =vrB ma+es an incision at the fifth phosphodiester bond on the 1E side of the lesion. This is followed by a =vrC mediated incision at the eighth phosphodiester bond on the DE side. The resulting .? to .1 nucleotide fragment is removed by =vrD helicase. The short gap thus created is filled in by D<& polymerase 2 and D<& ligase.

2n humans and other eu+aryotes, the enzyme system hydrolyzes the si)th phosphodiester bond on the 1E side and the twenty second phosphodiester bond on the DE side, producing a fragment of ?B to ?A nucleotides. #ollowing the dual incision, the e)cised oligonucleotides are released from the duple) and the resulting gap is filledJby D<& polymerase 2 in ligase seals the nic+. Direct Repair -everal types of damage are repaired without removing a base or nucleotide. The best characterized e)ample is direct photoreactivation of cyclobutane pyrimidine dimers, a reaction promoted by DNA photolyases. %yrimidine dimers result from an ultraviolet light@induced reaction, and photolyases use energy derived from absorbed light to reverse the damage. %hotolyases generally contain two cofactors that serve as light absorbing agents, or chromophores. One of the chromophores is always #&D!L. 2n . coli and yeast, the other chromophore is a folate. . coli and D<& polymerase

in humans. D<&

.. & blue light photon 31>> to D>> nm wavelength$ is absorbed by the (T!#poly8lu, which functions as a photoantenna. ?. The e)citation energy passes to #&D!L in the active site of the enzyme. 1. The e)cited flavin 3M#&D!L$ donates an electron to the pyrimidine dimer 3shown here in a simplified representation$ to generate an unstable dimer radical. 0. Electronic rearrangement restores the monomeric pyrimidines, and D. The electron is transferred bac+ to the flavin radical to regenerate #&D!L.