BACKGROUND:
Cleaning of toilets with toilet cleaners is very necessary to protect our health. It is important to understand difference between cleaning and sanitizing. Cleaning is defined as "the systematic application of energy to the surface or substrate, with the intention of removing dirt. Sanitizing is the process of reducing the number of microorganism on the surface to a safe level. Infections are spread, by germs and bacteria. The toilet, like bathroom is a humid area of the house due to the amount of water in the cistern, and as we know germs love a humid environment to breed in. Bacteria in the toilet absorb organic waste and release gases which of bad smell. Every time if we flush the toilet they projected into the air up to l.Smeters high. 2.5 -billion People -almost two fifth of the world's population do not have access to adequate sanitation. It's also spread different types of diseases.
There are many sanitizer cleaners commercially available in the market like Phenol, Harpic, Lysol, Domestor etc. These sanitizers contain a lot of chemicals, acids and other ingredients which are harmful to human beings and environment. These acids clean but degrade the life period of ceramic sanitizers and causes diseases.
Composition of some commercial products: HARPIC: HC1 -10%'as the active ingredient and butyl oleylamine. LYSOL: Ethanol 1-3%, Isopropyl alcohol- 1-2 %, p-chloro-o-benzylphenol- 5-6%, o-phenyphenol 0.1%, potassium hydroxide 3-4%, Alkyl-50%,^ dimethylbenzyl ammonium saccharinate 0.10%, lactic acid , hydrogen peroxide.

UREASE: Urease functionally belongs to the super family of Amidohydrolases and Phosphotriesterases. It is an enzyme that catalyzes the hydrolysis of urea into carbon dioxide.and ammonia
(NH2)CO + H20 ->C02 + 2NH3 More specifically, urease catalyzes the hydrolysis of urea to produce ammonia and carbamate. Carbamate produced is spontaneous degraded to produce ammonia and carbonic acid. Urease activity tends to increases PH in environment. Urease found in numerous bacteria, fungi, algae, Jack bean. Optimum PH =7.4 Inhibitors heavy metals (like: lead)
Commercially-available liquid toilet bowl cleaning compositions generally incorporate-, mineral acids such as hydrochloric, sulfuric or phosphoric acid; A typical product might contain about 5-35% hydrochloric acid. In response to the danger associated with such liquid acid compositions, such as spillage onto substrates other than those intended for cleaning, skin burns or irritation, deleterious effects upon metals, accidental ingestion and the like compositions formulated as mixtures of solids were developed. These compositions incorporated acidic active ingredients such as sodium bisulfate, sulfamic acid, oxalic acid or citric acid in » combination with surfactants, disinfectants, dyes and perfumes. Solid compositions are considered to be safer to use. However, solid toilet bowl cleaning compositions present the problem of over- or underuse due to measuring difficulties.
Several patents describe cleaning compositions; however, each of them has several shortcomings. However, this composition does not effectively clean and

dissolve scale nor is it able to cling to the vertical walls of the bowl. This material is a slow acting cleaner and does not facilitate the active scouring of a bowl.
Thus, there is a. need in the art. for a liquid toilet cleaner that is-faster acting,
safer and thicker than previous compositions. The cleaner must be a quick and
effective means of cleaning a toilet bowl and not dissolving lime scale. It must not
be corrosive to delicate surfaces or initially irritating to the user. It must also cling to
the vertical walls of the bowl and maintain sufficient contact time. It also should be
safe to ecosystem. '
References:
1. US4330626A- Methods of preparing high-activity, low-bacteria,Urease enzyme. _
2. US 3409508 A- Methods for analysis of urea in biological fluids.

3. DONALD D.VAN SLYKE AND GLENN E. CULLEN, THE MODE OF ACTION UREASE AND OF ENZYMES IN GENERAL (From the Hospital of The Rockefeller Institute for Medical Research).
4. Robert L. Blakeley, Edwin C.Webb, Burt Zerner Biochemistry (1969), JACK BEAN UREASE (EC 3.5.1.5.) A NEW PURIFICATION AND RELIABLE RATE ASSAY, 8(5), ppl984-1990
5. NICHOLAS E.DIXON, CARLO GAZZZOLA, ROBERT L. BLAKELEY, BURT ZERNER (1975), JACK BEAN UREASE (E.C. 3.5.1.5.) METALLOENZYME. SIMPLE BIOLOGICAL ROLE FOR NICKEL, J. Am.Chem.soc.,97(14), pp 4131-4133
6. U.S. Pat. No. 5,322,635 to Hieatt is directed to a soap composition.

, 7. U.S.Pat.No 4,269,723 to Barford et al. is directed" to a particulate material
and solid tablets that slowly dissolve in the cistern of a toilet bowl. Claims:
1. A novel biotechnologicai toilet cleaner containing Urease, sanitizer base, buffer and thickening agents.
2. As mentioned in the claimed, Urease (1 to 20%) obtained from Jack bean or Bacillus sps bacteria.
3. As mentioned in claimed, the sanitizer base is ATX (5-25%) and SDS (1-10%)
4. As mentioned in claimed , the buffer is 0.1M Phosphate or Acetate (pH 2-8)
5. As mentioned in claimed, the thickeners are CMC (1-10%), NaCl (1-10%)
6. A novel Biotechnologicai toilet cleaner prepared as claims 1 to 5 can be used for cleaning toilet bowls, toilet, sinks, washbasins, etc. It should be applied on surface, allow for 30minutes and wash with brush and water.
Description:
1. The current invention relates to solve the problem of corrosive toilet cleaners. In one aspect, the present invention relates to biotechnologicai toilet cleaner containing Urease (1 to 20%), ATX (5-25%),'SDS (1 to 10%), NaCl (1 to 10%) and CMC (1 to 10%)
2. In another aspect, present invention relates to preparation of toilet cleaner by taking all of above components in their required concentrations.
3. In another aspect, the method of using the toilet cleaner is to apply the cleaner on site of application i.e., Toilet bowls, wash basins, toilet floor and

walls, urinals and sinks etc., allow for 30 minutes and wash with brushed and water.
4. In another aspect, the invention is non-corrosive and made-up of Urease enzyme and other components.
5. Toilet apparatus are not damaged with biotechnological toilets cleaner as it is happened with normal chemical toilets cleaners.
, Summary: Problems to be solved:
1. In all civilized societies, toilet cleaners are being used to clean the dirt and deodour the toilets, toilet bowls, sinks etc.
2. High concentrations of acids making commercial toilet cleaners corrosive and damaging toilet apparatus.
3. Commercial toilet cleaner are expensive.
Means of solving problem:
1. The present invention relates to biotechnological preparation of toilet cleaner.
2. Urease of Jack bean and Bacillus species was extracted in buffer and used.
3. Low concentration of ATX, SDS were used for dirt removal, NaCl and CMC were used to thickening.
Advantages of Invention:
1. Urease enzyme is used to deodorize urea and nitrogenous waste and anti-bacterial activity.
2. Very low concentration of chemicals were used, hence not a corrosive.

Example 1:
Extraction of enzyme: Urease:
Jack-bean seeds are collected; thin peels present on seeds are removed and dried completely. These seeds are grinded to fine powder. The phosphate buffer 0.1M pH7 is prepared. 20gms of jack bean powder is added in 100ml phosphate buffer and kept on magnetic stirrer for an hour. After an hour the solution is filtered using filter paper. The liquid which is collected is urease enzyme. It is checked for activity by ninhydrin test. It is found positive for this test Example 2:
Best toilet cleaner preparation: Ingredients:
Urease - 10% (aqueous containing 1 Omg/ml) ATX -20% SDS -1% NaCl -1 %. CMC -1 %
Preparation: Urease was extracted from.Jack beans which were powdered finely and 20 grams jack bean powder was added to O.lMphosphate buffer and allowed for stirring on magnetic stirrer for 60 minutes.
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Now 200ml of classical ATX (sanitizer base) is taken and diluted for 5 times and made it to 1 liter. To this suspension add 100ml of Urease (lOmg/ml protein), 10 grams SDS and mixed it properly. This prepared sample is very much diluted and it cannot stop on toilets as it lacks stickiness. So to get rid of this problem 10 grams of Methyl cellulose and 10 grams of Sodium chloride is added and mixed properly and the solution now becomes a perfect sticky sanitizer.
Example3 Anti-microbial activity:
Anti-microbial activity of Urease:
Urease, Biotechnological toilet cleaner and commercial toilet cleaner (Harpic) were evaluated by well diffusion method against toilet pathogenic bacteria like E.coli , Klebsiella , Pseudomonas, Entervbacter , Bacillus ,Streptococcus, Clostridium etc.
Luria agar plates were prepared and 12mm wells were prepared with sterile dweller. Cultures were grown in Luria broth for overnight at 37°C 100 RPM to get active culture. The cultures were spreads on wells prepared Luria agar plates with sterile cotton swab. lOOmicro liter each Urease, biotechnological toilet cleaner, commercial toilet cleaner and standard antibiotic (10|igAmphicilin) were added and incubated at 37° c for overnight.
Clearance zone was observed as anti-microbial activity. Biotechnological toilet cleaner was effective antimicrobial against all the tested microorganisms whereas Harpic is effective against gram negative bacteria.
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Example 4
Method of usage: It is used and tested activity on toilet bowls and toilets. 1 liter of
sample is good enough for 10 toilet bowls washes. The solution is poured on
different bathrooms, lavatories and allowed for 20 min. Then washed with the help
of toilet brush and water. It is effective in dirt removal and deodouring. It is also
compared with the present sanitizers in the market. Biotechnological toilet cleaner
was better cleaning but taking 10 minutes more than commercial Harpic. It takes
time as - it contain enzyme and enzyme activity depends upon environmental
conditions.