Sucrose Based Thermal Stabilization Of Enzymes Biology Essay

Invention relates to a method for saccharose based thermic stabilisation of enzymes for breakage of biopolymer harm in gas and oil Wellss. The said enzyme includes Amylase and/or a Biopolymer Hydrolase Enzyme mix of Cellulase and/or Hemicellulase-Mannanase enzymes. The method comprises measure of fixing Thermostable Enzyme Treatment Fluid by uniting the Amylase and Hydrolase enzymes with Sucrose and seawater and a wetting agent, widening into an oil or gas or H2O bring forthing reservoir via a good bore with high down hole temperatures, utilizing a drill twine or deployed utilizing a coiled tubing or bullheaded into the breaks, while shooting the intervention fluid below or above the break force per unit area and close off the contents for a period of clip, leting to respond and disintegrate the Biopolymer based formation harm or filter cake wholly and later removed by normal blushing techniques known mode.

The Field of innovation

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The innovation relates to method for saccharose based thermic stabilisation of enzymes for breakage of biopolymer harm in gas and oil Wellss. More peculiarly the method of the present innovation is applicable to the production of oil, gas or H2O from Wellss drilled into belowground reservoirs where a Biopolymer based formation harm needs to be expeditiously disintegrated by Hydrolase enzymes, by virtuousness of Thermally Stabilizing it, despite the high downhole temperatures, which would otherwise wholly or partly deactivate the enzyme rendering it ‘s dynamicss unpredictable and inefficient.

Description of the Prior Art:

Since the formation harm can be in such country which is near to the oil bring forthing openhole, the same could mostly consequence production. Therefore, such Biopolymer Filter cake based harm demands to be efficaciously removed to significantly increase production of Hydrocarbon or H2O in H2O bring forthing Wellss and injectivity of such Wellss.

Though, such Biopolymer based formation harm needs to be removed to increase the flow of production fluids from the formation, it can non be easy flushed out of the formation being about indissoluble in aqueous fluids,

As a consequence, remotion of such filter bar warrants either intervention by strong oxidizers, like persulfates or strong acids, both of which are highly risky for operation and environment.

In the yesteryear, with a position to excite an under-producing well, frequently a rough Inorganic Acids ( e.g. HCL ) has been used to free and liquefy the harm and take quickly. However, it has been observed that such patterns have many disadvantages, which includes and non limited to, organizing of worm holes leaking off intervention fluid, eating tubings and stop uping screens, triping localised breakage, etc. Particularly for deviated and horizontal Wellss, the intervention fluid would leak off through worm holes incorporating strong acids or oxidizers, and pollute gas or H2O beds. Such leaking off besides cut down the zone of intervention of formation harm and via media productiveness.

Strong acids are non merely risky to manage, they are besides caustic to tubing and equipments, ensuing in Sludging and polluting petroleum and clogging screens, which is an expensive matter to clean. Corrosion inhibitors are normally toxic in nature.

Since Calcium Carbonates are frequently used as a weighting agent in the boring clay preparation, the Filtercake would practically dwell of a combination of Carbonate and the Biopolymer matrix. Carbonate mulcts generated during boring of carbonate stones may besides be present in the filter bars.

Filter bars signifier as the gel fluids are pumped into the subterraneous formation and some portion of the fluid leaks into the little stone pores of the formation, go forthing behind the supermolecules of biopolymer gel on the stone surface, organizing a comparatively impermeable bed. Those Biopolymers which does non organize bars, still increases viscousness on localisation, which acts similar to a filtercake, barricading off production fluid. Since the Filtercake is a concentrated retentate buildup of the fracturing liquid, it frequently contains high densenesss of polyose, U.S.Pat. No. 5,247,995 citations SPE Paper 21497 bespeaking that they can incorporate up to about 48 % polysaccharide versus about 4 % in fracturing fluids, which no uncertainty, is significant.

U.S. Pat. No. 5,165,477, to Shell, et. al. , and assigned to Phillips Petroleum Co. which describes a method of taking used boring clay of the type consisting solid stuffs including at least one polymeric organic viscosifier from a well dullard and parts of formations next thereto comprising: shooting a well intervention fluid consisting an enzyme capable of quickly enzymatically degrading polymeric organic viscosifier into good ; and leting enzyme to degrade polymeric organic viscosifier and good intervention fluid to scatter used boring clay. In this innovation, Shell adds the enzyme to a viscosifier.

U.S. Pat. No. 5,881,813, to Brannon, et. al. , and assigned to Phillips Petroleum Co. describes a method for bettering the effectivity of a well intervention in subterraneous formations consisting the stairss of: shooting a clean-up fluid into the well wherein the clean-up fluid contains one or more enzymes in an sum sufficient to degrade polymeric viscosifiers ; reaching the well bore and formation with the clean-up fluid for a period of clip sufficient to degrade polymeric viscosifiers therein ; executing a intervention to take non-polymer solids that may be present ; and taking the non-polymer solids in the well to better productiveness or injectivity of the subterraneous formation.

U.S. Pat. No. 5,247,995, to Tjon-Joe-Pin, et. al. , and assigned to BJ Services, which describes a method of increasing the flow of production fluids from a subterraneous formation by taking a polysaccharide-containing filter bar formed during production operations and found within the subterraneous formation which surrounds a completed good bore consisting the stairss of leting production fluids to flux from the well bore, cut downing the flow of production fluids from the formation below expected flow rates and explicating an enzyme intervention by intermixing together an aqueous fluid and enzymes. The enzyme intervention is pumped to a desired location within the well bore and the enzyme intervention is allowed to degrade the polysaccharide-containing filter bar, whereby the filter bar can be removed from the subterraneous formation to the well surface.

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U.S. Pat. No. 6,110,875, to Tjon-Joe-Pin, et. al. , and assigned to BJ Services, describes a method for degrading xanthan molecules consisting the measure of reaching the molecules with xanthanase enzyme composite produced by a dirt bacteria bearing the ATCC No. 55941 under conditions such that at least a part of the molecules are degraded.

U.S. Pat. No. 6,936,454, to Kelly, et. al. , and assigned to North Carolina State University, which describes a composing consisting an stray mannanase enzyme that hydrolyzes ( 3-1,4 hemicellulolytic linkages in galactomannans at a temperature above 180A° F. and that is basically incapable of degrading the linkages at a temperature of 100A° F. or less.

US Pat. No 4,617,662, to Curtis J, et. al. , and assigned to Miles Laboratories, Inc. disclosed is a method for heightening the thermic stableness of microbic alpha-amylase. The method involves adding a stabilising sum of an amphiphile to the enzyme in its aqueous solution. Besides included within the range of the innovation is the stabilised alpha-amylase preparation and its usage in the liquefaction of amylum.

U.S. Pat. No. 4,284,722, to Masaki Tamuri, et.al. , and assigned to CPC International Inc.claims a heat and acerb stable alpha-amylase derived from an being of the species Bacillus stearothermophilus.

U.S. Pat. No. 4,497,897 to Jens H. Eilertsen, et. al. , and assigned to Novo Industri A/S, disclosed a method for heightening the shelf life during storage of peptidase from Subtilisin Carlsberg which involves the add-on of Ca ion and a H2O soluble carboxylate selected from the group of formate, ethanoate, propionate and mixtures thereof to a solution of the enzyme.

U.S. Pat. No. 4,451,569, to Setsuo Kobayashi, et.al. , and assigned to Toyo Boseki Kabushiki Kaisha, describes a stable enzyme composing consisting glutathione peroxidase and at least one stabilizer compound selected from the group dwelling of pentoses, hexoses, penthahydric sugar intoxicants, hexahydric sugar intoxicants and disaccharides.

K.Samborska et al. , Journal of Food Process Engineering 29 ( 2006 ) 287-303. study that among all stabilising compounds investigated, sucrose exhibited the largest protective consequence on tried Amylase enzyme. The denary decrease clip of a-amylase activity increased by 33.9 times when 420 mg/mL of saccharose was added to the environment. When the same concentration of trehalose was used, the D-value increased by 6.4 times compared to the value in the buffer system. The nOH provided in the enzyme

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solution could non be related to the D-values for the enzyme thermic inactivation, intending that the enzyme heat stableness was non dependent on the nOH.

JC Lee and SN Timasheff et al. , J. Biol. Chem. , Vol. 256, Issue 14, 7193-7201, Jul, 1981, reports that the consequences from the protein-solvent interaction survey indicate that saccharose is preferentially excluded from the protein sphere, increasing the free energy of the system.They study that Thermodynamically this leads to protein stabilisation since the unfolded province of the protein becomes thermodynamically even less favourable in the presence of saccharose.

Once subjected to High Temperatures that could otherwise deactivate enzymes in downhole conditions, effectivity of a Filtercake breakage Hydrolase enzyme system can be established in the research lab. This can be done, either by look intoing the addition of filtration rate through a Biopolymer based filter-bed of similar composing prepared and cured in the lab or merely by gauging the cut downing sugar produced as a hydrolysis merchandise of the Biopolymer in the bed.

It has been reported that although enzymes potentially offer a figure of advantages over conventional chemical accelerators, they are by and large unstable in utmost conditions and they get deactivated quickly by heat and other environmental alterations such as alterations in pH and ionic disA¬balance. Since the active site of the enzyme consists of amino acids brought together merely in the native 3-dimensional construction, an unfolded enzyme loses its catalytic activity.

Therefore, thermally strengthening and protecting the Hydrolase enzymes to present an efficient Enzyme Hydrolysis at highly high downhole temperatures non merely bears commercial value of Enzyme dose in intervention fluid preparation, it besides provides higher predictability of the Enzyme kinetic rate in downhole conditions, conveying about easiness in operation, planning and computation.

It is an object of the present innovation to supply a simple and effectual method for Thermal Fortification of the Hydrolase enzymes by stabilising it even at highly high downhole temperatures of an belowground reservoir, for an efficient and predictable Enzymatic hydrolysis of the Biopolymers.

It is a peculiar object of the present innovation to supply simple and effectual methods for the efficient remotion of filter bars from downhole conditions with comparatively high temperatures, above 100 Deg C, which could potentially deactivate most enzymes with incubation.

Another object of the present innovation is to supply a thermic munition for the enzyme proteins for a more predictable operation of the enzymes in hydrolysing the Biopolymers even at non so high temperatures e.g. below 100 Deg C, but more than 60 Deg C, so that Biopolymer based harm can be

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easy removed and matrix permeableness can be increased for a better production.

It is a farther object of the present innovation to supply methods which are non harmful for the environment and easy to manage by the operators without any sum of jeopardies.

Summary of the Invention:

The present innovation provides a method of Thermally Stabilizing Hydrolase enzymes for pumping into a subterraneous formation bearing high downhole temperatures, along with other ingredients of the Treatment Fluid, for effectual remotion of Biopolymer based filter bar, without the Enzyme acquiring De-activated. Efficient remotion of filter bar increases the permeableness of the formation or the break, exciting the well to bring forth at a higher rate.

The method relates to Hydrolase enzyme chosen based on the nature of the Biopolymer substrate, e.g. Amylase for Starch substrate, Cellulase for a CMC substrate or Mannanase for Guar derivative based harm substrate, to Thermally Stabilize the enzyme and guarantee that it works at an efficient kinetic rate, even when subjected to high downhole temperatures. The range of this innovation peculiarly addresses oil, gas or H2O bring forthing Wellss with high downhole temperatures runing between 80 to 130 Deg C, that is 176 to 266 Deg F.

Consequently the innovation provides a method for saccharose based thermic stabilisation of enzymes for breakage of biopolymer harm in gas and oil Wellss, said enzyme includes Amylase and/or a Biopolymer Hydrolase Enzyme mix of Cellulase and/or Hemicellulase-Mannanase enzymes, the said method comprises measure of fixing Thermostable Enzyme Treatment Fluid by uniting the Amylase and Hydrolase enzymes with Sucrose and seawater and a wetting agent, widening into an oil or gas or H2O bring forthing reservoir via a wellbore with high downhole temperatures, utilizing a drill twine or deployed utilizing a coiled tubing or bullhead into the breaks, while shooting the intervention fluid below or above the break force per unit area and close off the contents for a period of clip, leting to respond and disintegrate the Biopolymer based formation harm or filter cake wholly and later removed by normal blushing techniques known mode.

Detailed Description of Invention:

The present innovation provides a method for handling an belowground reservoir, which method comprises presenting into the reservoir a intervention

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fluid comprising, dissolved or dispersed in H2O, a Hydrolase enzyme or a combination of Hydrolase Enzymes aimed at a complex Biopolymer substrate, and a Sugar to protect the Hydrolase enzyme from Thermal Deactivation in high downhole temperatures.

The method of the innovation fortifies and thermally Stabilizes Hydrolase Enzymes, enabling these to remain active at high downhole temperatures and partly or wholly disintegrate Biopolymer Matrix of filter bar formed due to utilizing polysaccharide-containing boring fluids, in subterraneous formations. Without suited stabilisation or munition, the Hydrolase enzymes would be wholly or partly deactivated in the Treatment fluid, and non execute as predicted, in an efficient mode.

When a Biopolymeric Viscosifier based boring clay is used in a subterraneous reservoir, a filter bar signifiers on the stone matrix by filtrating the aqueous fluid from the polymer suspension, through the little stone pores. This filtercake contributes to reduced permeableness of the formation and slows down production well. This Filtercake of Biopolymeric sedimentations consists chiefly the Long concatenation Polysaccharide Biopolymers implanting the carbonate weighting agents, frequently used in H2O based clay. These Polysaccharides can be Starch or Amylum, Xanthan Gum and derived functions, Cellulose and derived functions like Carboxymethyle Cellulose or Guar Derived functions like Hydroxymethylguar or Hydroxypropylguar.

Starch based Biopolymeric Viscosifiers in formation harm are nil but glucose polymers linked together by the alpha-1,4 and alpha-1,6 glucosidic bonds. Because of the being of two types of linkages, the alpha-1,4 and the alpha-1,6, different conformational being are possible for amylum molecules. An un-branched, individual concatenation polymer of 500 to 2000 glucose fractional monetary units with merely the alpha-1,4 glucosidic bonds is called amylose, while the presence of alpha-1,6 glucosidic linkages consequences in a bifurcate glucose polymer called amylopectin. The grade of ramification in amylopectin is about one per 25 glucose units in the unbranching sections.

Another Biopolymer used as clay viscosifier is Cellulose, the most abundant biopolymer on Earth. Cellulose consists of D-glucopyranose monomer units bound by P ( 1- & gt ; 4 ) glycosidic linkages. The cellulose molecule forms a linear, about to the full extended concatenation with a double prison guard axis on which consecutive glucose residues are rotated 180A° comparative to each other and the glycosidic Os point instead up and down.

Another Biopolymer based gelling agent Guar gum-Galactomannan is a high molecular weight saccharide polymer derived from the natural seed of cluster bean works ( Cyampopis tetragonolobus ) . Part of the seed is Hull ( 14-17 % ) , Endosperm ( 35-42 % ) , and source ( 43-47 % ) . Guar gum is a polysaccharide consisting of a mannose anchor with a galactose side concatenation. The

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brain sugar is indiscriminately placed on the mannose anchor with the mean ratio 1:2 of brain sugar to mannose.

Though enzymes offer a figure of advantages over conventional chemical accelerators, they are by and large unstable in utmost conditions and are inactivated quickly by heat and other environmental alterations such as alterations in pH and ionic disbalance. Since the active site of the enzyme consists of amino acids brought together merely in the native 3-dimensional construction, an unfolded enzyme loses its catalytic activity.

Therefore, an efficient Enzyme Hydrolysis non merely bears commercial value in footings of Enzyme dose in intervention fluid preparation, it besides provides higher predictability of the Enzyme ‘s Kinetic rate in downhole conditions.

The method in this innovation includes an add-on of sugars to aqueous solutions of the Hydrolase enzymes in the Treatment fluid preparation, therefore beef uping the hydrophobic interactions among nonionic amino acid residues. These interactions, together with H bonds and ionic and van der Waals interactions, are indispensable to keep the indigen, catalytically active construction of the enzyme. Therefore, the strengthened hydrophobic interactions make protein supermolecules more stiff, and hence more immune to thermal flowering, which would otherwise render the Hydrolase enzymes inactive in downhole conditions, when the intervention fluid is pumped and shut in for hours at highly high temperatures.

This method of enzyme stabilisation in the Treatment Fluid preparation in the presence of sugars in aqueous media do non alter the protein conformation, but influence the physicochemical belongingss of the system such as the dissolver construction, ensuing in protein stabilisation. The solvent composing in the immediate sphere of the protein is different from that of the majority dissolver and the difference is a map of the concentration of co-solvent. This co-solvent added to the enzyme aqueous solution is excluded from the protein sphere and increases the Free Energy switching the Thermodynamic Equilibrium towards the native province.

In the peculiar method, the Sugar is Sucrose, a disaccharide of glucose and fruit sugar with an a ( alpha ) 1,2 Glycosidic linkage and with molecular expression C12H22O11 The concentration of Sucrose in the Treatment fluid should be sufficient to achieve Thermal Stability of the 3-D construction of the Enzyme Protein in high downhole temperatures. The concentration of sugar incorporated into the intervention fluid of the present innovation will be from 0.1 % w/v but may be up to 5 % w/v ( 1 to 50 kilograms per M3 ) . In general it has been found that 0.2 % to 4 % w/v ( 2 to 40 kilograms per M3 ) sugar when used in combination with the Hydrolase enzymes and 2 % seawater or sea H2O, well fortifies the enzymes against rough downhole conditions.

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When the Hydrolytic activity of a amylum degrading Enzyme is fortified against high downhole temperatures as per this method, Gelatinized and polymerized amylum of the formation harm would be actively attacked by the Enzyme constituents of the intervention fluid in contact. Depending on the comparative location of the bond under onslaught as counted from the terminal of the concatenation, the H2O soluble merchandises could be dextrin, maltotriose, malt sugar, and glucose, etc. The specificity of the bond attacked by alpha-amylases depends on the beginnings of the enzymes e.g. bacterial or, fungous. Normally a Bacterial alpha-amylase randomly attacks merely the alpha-1,4 bonds, and liquefies gelatinized amylum of the Filtercake of the harm, foremost cut downing its viscousness by cutting down concatenation length and eventually rendering it soluble in H2O.

Amongst the major types of Amylase enzymes, this method of Thermal Stabilization of Treatment Fluid Enzymes relates to Alpha or Endo Amylases, which are 1,4-a-D-glucan glucanohydrolase. The Thermally Stabilized Endo-Amylase, by moving at random locations along the amylum concatenation of the formation harm in contact, will interrupt down long-chain saccharides, finally giving maltotriose and malt sugar from amylose, or maltose, glucose and “ bound dextrin ” from amylopectin. Because it can move anyplace on the substrate, a-amylase tends to be faster-acting than 3-amylase or Glucoamylases, and therefore preferred for this method.

Depending on the type of amylum used in the formation harm, Beta or Exo Amylases, 1,4-a-D-glucan maltohydrolase would work from the non-reducing terminal and catalyse the hydrolysis of the 2nd a-1,4 glycosidic bond, spliting off two glucose units ( malt sugar ) at a clip. Finally, the Amyloglucosidase or Glucoamylases constituent of the intervention fluid would split a ( 1-6 ) glycosidic linkages and the last a ( 1-4 ) glycosidic linkages at the non-reducing terminal of amylose and amylopectin, giving glucose.

When thermally protected as per this method against temperature daze at downhole conditions, Cellulolytic Hydrolase enzymes or, cellulases, would hydrolyze -1, 4- glycosidic linkages in cellulose Biopolymer of the formation harm. Three different cellulolytic activities can be used in the procedure, e.g. Exoglucanases ( 1, 4- -D-glucan cellobiohydrolase ) hydrolyse cellulose from the free concatenation terminals, bring forthing chiefly cellobiose and are called Exo cellulase or cellobiohydrolases. Endoglucanases ( 1,4- D-glucan-4-glucanohydrolase ) on the other manus onslaught randomly internal linkages within the cellulose concatenation, P -Glucosidases eventually hydrolyse little oligomers, chiefly trimers and dimers, to H2O soluble monomers.

When thermally protected as per this method against temperature daze at downhole conditions, the Guar derivative biopolymer Hydrolase enzymes are Galactomannanase or Mannanase type of Hemicellulase that will assail the galactomannosidic and mannosidic linkages in the cluster bean residue, interrupting the molecules into monosaccharose and disaccharide fragments, which are soluble in H2O and renders the clay harm to be easy removable.

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These Hydrolase enymes specifically hydrolyse the ( 1,6 ) -.alpha.-D-galactomannosidic and the ( 1,4 ) -.beta.-D-mannosidic linkages between the monosaccharide units in the guar-containing filter bar severally.

In the peculiar method, sufficient sum of Hydrolase enzyme should be present in the Treatment fluid preparation, to be able to expeditiously disintegrate and slow the stiff filter Biopolymer matrix of the formation harm. Depending on the type of Biopolymer used in mud preparation, an Amylase, a Cellulase or a Hemicellulase enzyme should be chosen to work independently or in a combination, whereas the combination of enzymes should be in the same ratio of the biopolymer substrate used in mud preparation. Typically a individual enzyme or multiple enzymes together of entire 0.1 to 5 % w/v or, 1 to 50 kg/m3 of the Treatment fluid preparation has been found to be suited for an effectual disintegration of the Biopolymer Matrix when Thermally fortified as per this method.

As per the method, the intervention fluid is prepared by fade outing the Thermal Stabilizer Sucrose in suited H2O, like metropolis H2O or produced brine H2O or sea H2O and eventually the Enzymes are added to the commixture armored combat vehicle. Since no rough chemicals are used in this method, tank stuff can be mild steel or a polymer like HDPE. Subsequently other chemical additives like wetting agents, chelating agents, antifoam or biocides can be added as are normally used in the oil industry. A suited biocide will command growing of unwanted bugs in the intervention fluid during pumping and shut in hours.

The assorted fluid is introduced into the belowground formation via injection or production Wellss and for freshly drilled wells the fluid may present through the drill threading utilizing the clay pumps or spotted by utilizing Coiled tube or Bullheading to reach the zone of formation harm or filter bars. Depending on formation and reservoir belongingss, the intervention fluid would be introduced at force per unit area below or above break force per unit area.

Depending on the openhole or near wellbore interventions, volume of intervention fluid will typically be at 120 % to 200 % of the openhole volume or good bore volume, accounting for the leak-offs and dead volumes.

Enzyme dynamicss being straight influenced by pH and Temperature conditions at downhole, the enzyme intervention is shut in the formation for a clip sufficient to expeditiously degrade the harm and as per this method will change between 45 proceedingss to a hebdomad, sooner 3 to 36 hours for an efficient harm remotion.

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The present innovation has the undermentioned peculiar advantages over the anterior art:

The method provides a simple manner to Thermally Fortify and maximise the benefits of Hydrolase enzymes for effectual remotion of Biopolymer based harm from a subterraneous formation, where downhole temperatures can be really high and may otherwise deactivate big parts of these enzymes.

Though enzymatic clay interrupting offers a figure of advantages over conventional chemical accelerators, there is every possibility of enzymes to suddenly retard or even halt working at high downhole temperatures. This makes it hard for the operator to judge the effectivity of enzyme in disassociating Biopolymer based formation harm, in a predictable Kinetic rate, and be after his operations consequently. However, this method of sugar add-on to the intervention fluid will beef up the hydrophobic interactions among non-polar amino acid residues and these interactions, together with H bonds and ionic and van der Waals interactions, would keep the indigen, catalytically active construction of the enzyme and non allow it be deactivated at comparatively higher temperatures.

An efficient Enzyme Hydrolysis would intend decrease of enzyme doses to achieve same degree of contact action rate or bar breakage, straight set uping economic sciences of such procedure.

Since the non reactive nature of this fluid preparation, it can be easy deployed for deep well stimulation every bit good as break face harm near the surface.

Sugar, being a nutrient class trade good produced from natural beginnings, is wholly environment friendly and at this degree, readily biodegrades when assorted in H2O or, land

The undermentioned illustration illustrates the innovation.

The Thermal stabilising consequence as per current innovation was tested by double method of Filtration rate addition and Reducing Sugar production for a scope of Treatment fluids with different concentrations of Thermally Stabilizing Sugar.

Blank filtration rate through perforated phonograph record with Whatman No 1 Filter-paper was checked at 2 Atmosphere Vacuum utilizing 2 % brine solution. The Disc was so damaged by go throughing clay incorporating Biopolymers like Starch, Xanthan gum, Calcium Carbonate burdening agents and Sodium Hydroxide under 2 Atmosphere vacuity. The filter bar was allowed to settle farther by incubating at 90 Deg C for 4 hours. The filtercake was so incubated with different intervention fluids at 105 Deg C for 12 hours. After incubation, the

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filtercakes were subjected to 2 % seawater flow at 2 atmosphere and Filtration rate tested and presented in Table 1.

The Residual filter bar and Filtration Permeates of each sample was collected individually & A ; assorted good. Reducing sugar production due to Enzymatic Hydrolysis was estimated by following method. The aliquoted samples ( 1.5ml ) were centrifuged for 15min at 10,000rpm. The supernatant was collected and centrifuged for 15min at 10,000rpm. The pellets were discarded and 700ul_ supernatant was taken from all the tubings in Boiling Tubes. To this, 50mL Sodium ethanoate buffer ( pH 4.8 ) and 1.2ml_ DNS ( Dinitrosalicylic Acid Solution ) were added and tubings were incubated for 30 min at room temperature. Subsequently, the tubings were put in Boiling H2O Bath and incubated at 95A°C for 15mins. The tubings were so transferred to a H2O bath at Room Temperature for 10min and so incubated at Room Temp for 10min. The curdled samples were so crushed good in their ain contents and to the full collected in Eppendorf tubings and centrifuged at 10,000rpm for 10min. Supernatant was collected and diluted with DM H2O ( as per below table 2 ) to acquire sufficient volume for Cuvette used in Spectrophotometer. Absorbance was measured at 540nm and calculated back utilizing the expression: Final O.D of sample = [ ( Vol of Supernatant + Vol of thining H2O ) /Vol of Supernatant ] * Measured O.D. The Reducing sugar production was observed as per Table 2.

Treatment Fluid Compositions

TFA1 composing:

Alpha Amylase Enzyme

Assorted Carbohydrase Enzyme

Sucrose

Seawater

DM Water

TFA2 composing:

Alpha Amylase Enzyme

Assorted Carbohydrase Enzyme

Sucrose

Seawater

DM Water

TFA3 composing:

Alpha Amylase Enzyme

Assorted Carbohydrase Enzyme

Sucrose

Seawater

DM Water

4 % 2 % 1 % 2 % Staying

4 % 2 % 3 % 2 % Staying

4 % 2 % 5 % 2 % Staying

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Table 1

Sample Brine Flow

Rate

through

Damage Brine Flow as % of

Blank after Enzyme

Treatment Crushability of Filter-cake

A-Mud Control 0 4 % Hard bar

B- Mud + TFA1 0 62 % Crushable bar

C-Mud + TFA2 0 75 % Easily crushable and mostly soluble bar

D-Mud + TFA3 0 93 % Easily crushable and Largely soluble bar

Table 2

Sample Measured OD Dilution Final OD Blanked OD

Supernat emmet ( MO Water ( Ml )

A-Mud Control I.052 650 350.079 0

B-Mud+TFA1.280 650 350.430 0.351

C-Mud + TFA2.376 900 600.626 0.547

D-Mud+TFA3.529 800 700.991 0.912

We Claim:

1. A method for saccharose based thermic stabilisation of enzymes for breakage of biopolymer harm in gas and oil Wellss, said enzyme includes Amylase and/or a Biopolymer Hydrolase Enzyme mix of Cellulase and/or Hemicellulase-Mannanase enzymes, the said method comprises measure of fixing Thermostable Enzyme Treatment Fluid by uniting the Amylase and Hydrolase enzymes with Sucrose and seawater and a wetting agent, widening into an oil or gas or H2O bring forthing reservoir via a wellbore with high downhole temperatures, utilizing a drill twine or deployed utilizing a coiled tubing or bullheaded into the breaks, while shooting the intervention fluid below or above the break force per unit area and close off the contents for a period of clip, leting to respond and disintegrate the Biopolymer based formation harm or filter cake wholly and later removed by normal blushing techniques known mode.

2. A method harmonizing to claim 1 wherein the belowground reservoir is a hydrocarbon reservoir.

3. A method harmonizing to claim 2 wherein the hydrocarbon is oil.

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4. A method harmonizing to claim 2 wherein the hydrocarbon is a gas.

5. A method harmonizing to claim 1 wherein the said Hydrolase Enzyme mix is a combination of an Amylase, Cellulase, Hemicellulase, Mannanase & A ; Galactomannanase.

6. A method harmonizing to claim 5 wherein all the said enzymes are assorted together or merely two of the said enzymes are chosen based on the nature of Biopolymer used in mud preparation.

7. A method harmonizing to claim 6 where the said enzymes are mixed in the same ratio of the Biopolymer substrates used in the clay preparation, on footing of declared Enzyme activity units.

8. A method harmonizing to claim 1 wherein the said Amylase is an Alpha Amylase enzyme that catalyses the endo-hydrolysis of 1,4-alpha-glycosidic linkages in amylum, animal starch, and related polyoses and oligosaccharides incorporating 3 or more 1,4-alpha-linked d-glucose units.

9. A method harmonizing to claim 1 wherein the Sucrose is a table sugar or saccharose and/or a disaccharide of glucose and fruit sugar with an a ( alpha ) 1,2 Glycosidic linkage and with molecular expression C12H22O11.

10. A method harmonizing to claim 1 wherein the Sucrose concentration is at least about 0.1 % w/v in the intervention fluid.

11. A method harmonizing to claim 1 wherein the said Amylase and Hydrolase Enzyme mix are liquid enzyme preparations.

12. A method harmonizing to claim 1 wherein the Biopolymer is Starch, Xanthan, Cellulose or Guar Derivatives.

13. A method harmonizing to claim 1 wherein the temperature of the formation bearing the Biopolymer based filtercake or mud harm of the reservoir is at least 60A° C. or higher.

14. A method harmonizing to claim 1 wherein the intervention fluid is shut off in the reservoir for at least 45 proceedingss.

15. A method harmonizing to claim 1 wherein the wellbore is perpendicular, deviated, inclined or horizontal.

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