We describe a recombinant host cell for the production of a compound of interest as well as isolated fungal promoter DNA sequences, to DNA constructs, vectors, and fungal host cells comprising these promoters in operative association with coding sequences encoding a compound of interest. The equation for calculation is shown below: Grams/Liter = 0.10(norm. 6 shows the activity for each immobilized enzyme as a function of the immobilized protein load. Laccases ( EC 1.10.3.2) are multicopper oxidases found in plants, fungi, and bacteria. The laccase activity measured by the three methods was greatly different. The enzyme had moderate thermal stability and was rapidly inactivated at 70 C. The maximum laccase activity using Firmiana platanifolia was approximately 93.01, 388.81, and 41.54 times greater than L. betulinus Han 851, S. rugosoannulata Han 1321, and A. heimuer Han 1333, respectively. So our percent enantiomeric excess is 86%. H2O2 - 50%) x (mL. H2O2 - 50%) x (mL. Immobilization of CotA laccase on MRGO-IDA-Cu 2+ nanosheets significantly enhanced the catalytic activity and stability of the enzyme. Abbreviations: ABTS 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), DOPA L-3,4-dihydroxyphenylalanine, DMP 2,6-dimethoxyphenol. No. 7 and and8 8 ). The edible fungi strains with short growing cycles originate from their high levels of secreted laccase activity. A molar extinction coefficient of 12 880 M 1 cm 1 for the quinoimine dye was used for calculation of the activity. Walter Steiner. Results obtained showed that, the enzyme formed by P. martensii NRC 345 expressed as specific activities, was detected mainly in the exocellular And let's round that to 86%. 2008) (Fig. This page is a summary of: Inconsistencies and ambiguities in calculating enzyme activity: The case of laccase, Journal of Microbiological Methods, December 2015, Elsevier, DOI: 10.1016/j.mimet.2015.10.007. How do you calculate laccase enzyme activity? All experiments were performed in triplicate, and the controls were subjected to the same parameters without the addition of the enzyme. The lowest values were assayed on the first day in decoloration of all dyes, and then, activities of laccase increased. Sample Laccase Activity may be calculated using the following equation: Sample Laccase Activity = M / (t x P) (pmol / (min x g)) = Units / g or mUnits / mg Where: M = Linear change in oxidized product concentration during t (in pmol) The lack of the second domain that is normally found in laccases raises several issues. Enzyme activity of samples with the lowest ionic strength (0.25 M) was defined as 100% and used for calculation of relative activity values. : 91-10-1. Laccase Acitivity The activity of the laccase was measured by monitoring the rate of oxidation of syringaldazine. Laccase (EC 1.10.3.2) is a cuproenzyme that oxidizes various types of phenols and similar aromatic compounds aromatic amines with the reduction of molecular oxygen to water, therefore, is used as a biocatalyst. Laccase is a key enzyme in the degradation of lignin by fungi. The kit is rapid, easy to use and high-throughput adaptable. Agrawal K, Verma P. 3 Biotech, 9(10):365, 21 Sep 2019 Cited by: 3 articles | PMID: 31588389 | PMCID: PMC6754822. We studied the laccase-catalysed oxygenation of methoxyphenolic food ingredients, such as 2-methoxyphenol (guaiacol) and 2,6-dimethoxyphenol (syringol), isomers such as 3- and 4-methoxyphenol, and 2,3-, 3,4- and 3,5-dimethoxyphenol. The solid horizontal line represents the residual activity at 50%. Mesosilica-coated ultrafine fibers for highly efficient laccase encapsulation, Wang SW, Chen W, He S, Zhao QL, Li XH, Sun JS, Jiang XY, Nanoscale, 6(12), 6468-6472. So that gives us our percent enantiomeric excess. (Ve/Vo) to calculate the size of the apoenzyme. The data obtained from the TAPPI method was based on the weight calculation, whereas the NREL data were obtained from HPLC analysis. For determination of laccase activities you mix your enzyme solution with an ABTS stock solution at a defined pH (use a buffer solution like e.g. Units/ml enzyme =. Effect of pH on laccase activity. The laccases are generally thought to follow Michaelis-Menten kinetics, and K m data are thus available as discussed above 37.Three datasets (named A, B and C) were used in the present study (Table 1).Dataset A is the smallest and is comprised of 11 well-known, structurally diverse laccase substrates with experimental K m values determined for Obtain the DA530nm/min. A simple method of assaying polyphenol oxidase is given: Add 2.5 ml of 0.1 M phosphate buffer pH 6.5, 0.3 ml of catechol solution (0.01 M) into cuvette and set the spectrophotometer at 495 nm. 1997). The relative enzyme activity was calculated by taking the laccase enzyme activity measured under the same reaction system without adding CuCl 2 solution was determined to be 100%. dear Christoph Andreas Gasser I may be wrong, but it looks like your calculations lead to MU/L instead of U/L thio.) As the real-time stability study progressed and once laccase activity was quantified, at each sampling moment, the remaining volume of samples (n) were pool (distinguishing by batch, by temperature, by sampling time) to determine the kinetics of the enzyme. You can use known control to see how reaction kinetics works. But no need to use standard. As it takes rate if reaction into account while measurin BioVisions Laccase Activity Assay Kit is a simple, one step, plate based assay to measure Laccase activity in various samples. For determination of laccase activities you mix your enzyme solution with an ABTS stock solution at a defined pH (use a buffer solution like e.g. M Other reactions without laccase involved express a higher reaction rate when the pH value increased. dear Pallavi Biswas are you sure that you don't have to multiply for 10^6 instead of divide? 1 mol =1000000 mol , you make the substitution and he Fungal laccases (EC 1.10.3.2) are multi-copper oxidases that oxidize a wide variety of substrates. 7.5 CALCULATIONS Units/ml enzyme = A530nm Sample = A530nm/min Test - A530nm/min Blank) (df) (0.001) (0.5) df = dilution factor 0.001 = the change in One unit (U) of laccase activity was defined as the amount of enzyme catalyzing the production of one micromole of coloured product per min per mL and expressed as number of Katals (1 mol of substrate conversion/s). Inconsistencies and ambiguities in calculating enzyme activity: The case of laccase. We don't need to worry about negative signs, so we can just take 27 and divide that by 31.5, and multiply it by 100, and we get 85.7. Tukayi Kudanga. Oxidation of ABTS was monitored by determining the increase at 414 nm. A 530nm Sample = A 530nm /min Test - A 530nm /min Blank) (df) (0.001) (0.5) df = dilution factor. 000 M1cm1.57A unit of activity was dened as the amount of laccase that oxidizes 1mol of ABTS per min. Laccase is a copper-containing polyphenol oxidase, so Cu 2+ is essential for the synthesis and activity of laccase. 0.001 = the change in A530nm/min. The assay mixture contained 5.1 mM ABTS, 1 mM sodium phosphate (pH 5), and a suitable amount of enzyme. The maximum laccase activity was found to be 186.41 0.15 U ml 1 from strain NITW715076_2 at 60 C after that the enzyme started losing its activity due to thermal instability. Determination of Reactions with ionic strength ranging from 0.25 to 2 M were performed and laccase activity was measured. Datasets used. The direct oxidation involves the oxidation of substrate to the corresponding radical as a result of direct interaction that occurs with copper cluster (Matera et al. And let's round that to 86%. CHARACTERIZATION OF LACCASE ACTIVITY PRODUCED BY Cryptococcus albidus Academia.edu uses cookies to personalize content, tailor ads and improve the user experience. Laccase activity was measured at 30C using 1 mM ABTS as the substrate in 100 mM sodium citrate buffer pH 5.0 were used to calculate kinetic constants (K m and V max) by the LineweaverBurk equation. The enzymatic activity at time 0 h was taken as comparison to calculate the relative enzyme activity of laccase in different time. The laccase activity was inhibited by NaN3 and NaF. Sample Laccase Activity may be calculated using the following equation: Sample Laccase Activity = Where: M = Linear change in oxidized product concentration during t (in pmol) t = t2 - t1 (in min) P = Sample protein content added to well (in g) A CotA laccase gene from B. subtilis WD23 was cloned and expressed in Escherichia coli. Purified laccase had an optimum H of 6.0 and was stable in the H range from 6 to 7. Walter Steiner. This link must help you http://www.sigmaaldrich.com/technical-documents/protocols/biology/enzymatic-assay-of-laccase.html Antioxidant activity assay based on laccase-generated radicals. Laccase activity levels in both cell-free filtrate (CFF) and cell-free extract (CFE) were determined. Laccase activity calculated using different equations. In the voltammetric method, laccase activity could be detected by putting platinum or glassy carbon electrode in Mcilvaine buffer. Laccase activity against HVA was expressed as H 2 O 2 equivalents (Lonergan et al. The laccase activity was determined according to a method reported by Paszczyski et al. Laccase activity has been expressed at colorimetric units ml-1 (CU ml-1). I am enclosing the paper for your reference also. Let me know if any clarity is needed. How to measure laccase activity using abts? How can I calculate laccase enzyme activity using ABTS? The equation for calculating laccase enzyme activity. The effect of pH on laccase activity was determined from pH 2.0 to 7.0 at 30 C. Analytical and Bioanalytical Chemistry, 2009. The highest 1).There are two weight-losses from 30 to 150 C and > 250 C for the FL, the first weight-loss is corresponding to the Despite extensive studies, the molecular basis for their diverse activity is The optimum Free to read. We also describe methods for expressing a gene of interest and/or A survey of specialist literature indicates that laccase activities in various fungi vary extremely, and range from 3.5 to 484,000 U L 1. Add little amount of the enzyme from the fermentation broth (Little amount 20-100ul is good to obtain good result because ABTS is highly reactive and the reading changes very fast). Start the stop watch immediately the enzyme is added and take the OD abs at every 1 min interval for 3 to 5 min. calculate laccase activity using change in OD @ 420 nm. Under the effect of direct current fields of 6 V voltage, samples were taken periodically in order to evaluate changes of enzyme activity in each region of enzyme solution (Figs. Reports indicate that the activity of this enzyme ranges from 3.5 to 484,000 U L(-1). What is the function of laccase corresponding gene products? As laccase (produced by Botrytis cinerea) can significantly alter the properties of wine, winemakers frequently use commercially available colorimetric kits and spectrophotometers to measure the activity of this enzyme in grapes, must and wine. Laccase Acitivity The activity of the laccase was measured by monitoring the rate of oxidation of syringaldazine. Laccase activity is commonly determined by monitoring spectrophotometric changes (absorbance) of the product or substrate during the enzymatic reaction. Most commonly used protocols for the measurement of laccase activity in soil or litter and methods applied for laccase activity assays in soil or litter. The oxidation rate of TEMPO by laccase varies at different pH conditions, and laccase activity is much higher at pH 4.5. Evaluation of laccase production by Ganoderma lucidum in submerged and solid-state fermentation using different inducers. : HY-W003972 Purity: 99.91%. So that gives us our percent enantiomeric excess. = 36,000 M-1 cm-1 = 36,000 L mol-1 cm-1, t = 3 min, d = 1 cm, Vt = 3 mL, Vs = 300 L, A = 0.50. The lowest values were assayed on the first day in decoloration of all dyes, and then, activities of laccase increased. By ABTS method, laccase activity could be detected in all KBS mediums with monoazo, disazo and triphenyl methane dyes (Figure 1). Laccase was assayed at different pH ranging from 3 to 9. Standard and sample solutions Laccase standard stock solution: Weigh the amount of laccase standard needed to obtain a laccase activity of 0.350 LAMU/ml and transfer the laccase to a 500-ml volumetric flask. Start the stop watch immediately the enzyme is added and take the OD abs at every 1 min interval for 3 to 5 min. In this assay, samples containing Laccase will oxidize the substrate to generate a colored product with a strong absorbance at 420 nm. We do not sell to patients. Molecular Mass Determination through SDS-PAGE. M (no. Laccase activity calculated using different equations. So let's get out the calculator, here. The laccase showed both syringaldazine and ABTS (2,2-azino-bis-ethylbenzthiazoline-6-sulfonic acid) oxidase activities and was activated by the addition of 1.7 Besides, the recovery of the heavy oil was significantly enhanced by 82.8%. Laccase. Size. Antioxidant activity assay based on laccase-generated radicals. The relative activity of laccase was increased by 16%, when CuSO 4 concentration was increased to 15 mM and Laccase was able to decolorize triphenylmethane dyes and azo-dyes. The purified laccase was active in broad pH range of 35 with optimum activity at pH 4.5 (Figure 5).The purified laccase has a broad temperature sensitive 3570C and the optimum temperature for the laccase was observed at 65C (Figure 6).Temperature kinetics of the laccase suggests that the enzyme 2,6-Dimethoxyphenol Chemical Structure. Sunnyvale, CA, USA). The laccase activity measured by the three methods was greatly different. 1) showed that the highest activity by the two yeast (Kluyveromyces sp.Dw1 and Pichia sp.Dw2) isolates was recorded at 192hr incubation time for all the carbon sources except for sucrose where Kluyveromyces sp.Dw1 had its highest laccase activity at 144hrs. As laccase (produced by Botrytis cinerea) can significantly alter the properties of wine, winemakers frequently use commercially available colorimetric kits and spectrophotometers to measure the activity of this enzyme in grapes, must and wine. Calculation is as follows: where is final volume of reaction mixture. x 34 (equiv. The ABTS oxidation assay is able to reliably detect the presence of 13.5 pg mL 1 or 0.199 10 12 mol mL 1 of T. versicolor laccase, which is three times more sensitive than the 2,6-dimethoxyphenol-based assay and more than 40 CotA protein is known as CotA laccase. Dear Pallavi Biswas, one publication using this protocol is for example: Zimmermann et al. (2011) Sorption-assisted surface conjugation: a way to s Activity calculation of laccase was followed as reported by Baltierra-Tejo et al. Although the used kits are based on electrochemically active substrates (such as syringaldazine and Tukayi Kudanga. it might help However, it is likely that this wide range is not due to the properties of the enzymes, but to inconsistencies in defining and calculating enzyme activity. The thermogravimetric analysis (TGA) curves are performed to examine the thermal properties of free laccase (FL), CKMB and laccase@CKMB with a constant heating rate of 10 C/min from 25 to 800 C under N 2 (Fig. How do you calculate laccase enzyme activity? (2014). (B) The t 1/2 (in min) at 70C of the twenty consensus variants and the OB-1 parental type. In all cases, lyophilized tissue was ground to a fine powder with a steel ball in a TissueLyser II (Qiagen, Venlo, Netherlands) for 2 min at 30 Hz and the tubes were weighed again to determine the dry biomass used for LAC activity calculation. In case of enzymatic profiling of blue laccase, its activity ranged from 0.04 to 464.3 U L -1 whereas in case of white laccase it ranged from 0.05 to 1404.7 U L -1 . Laccase was purified 3.95-fold with specific activity of 158 U/mg by ammonium sulfate precipitation, followed by gel filtration chromatography using Sephadex G-100 column. A locked padlock) or https:// means youve safely connected to the .gov website. Low molecular aromatic compounds similar in structure to lignin or lignin-degraded fragments can also induce and significantly increase laccase production, such as guaiacol, benzyl alcohol, resveratrol, vanillic acid, Tween and so on. Tukayi Kudanga. by using 2.8 m m DMP substrate in 100 m m acetate buffer pH 4.5. Fig. Purified laccase had an optimum H of 6.0 and was stable in the H range from 6 to 7. An enzyme showing alkaliphilic laccase activity was purified from the culture supernatant of Myrothecium verrucaria 24G-4. Laccase was purified 3.95-fold with specific activity of 158 U/mg by ammonium sulfate precipitation, followed by gel filtration chromatography using Sephadex G-100 column. Data Sheet SDS Handling Instructions. The concentrations of TEMPO and oxoammonium are monitored by EPR spectroscopy. Dear Elisabeth P Kristant ABTS is best and reliable method for Laccase activity at 420nm, ABTS concentration of the mixture should be 0.2 mM. Then The pH optimum of the laccase activity was 3.0 for ABTS and 5.0 for 2,6-dimethoxy phenol. These methoxyphenolic substrates generate unstable free radicals, w The relative activity was measured against control without the CuSO 4 which was taken as 100%. ABTS (1.6 mM, 0.5 ml) was added to the assay mixture and the formation of oxidation products monitored for 10 min. My question is related to Laccase enzyme activity. Do we need to use standard here. As in many research paper about standard is not mentioned. You can use known control to see how reaction kinetics works. But no need to use standard. The basic reactions catalyzed by laccase can be of two types: direct oxidation and in-direct oxidation. For measurement of extracellular laccase activity (ecLAC), the decanted media were stored at 20 C. Protein glycosylation, a major post-translational modification, plays essential roles in eukaryotic cells. The maximum enzyme activity was set as 100% for calculation of relative enzyme activity. Laccase (EC 1.10.3.2) is a cuproenzyme that oxidizes various types of phenols and similar aromatic compounds aromatic amines with the reduction of molecular oxygen to water, therefore, is used as a biocatalyst. 7.5 CALCULATIONS. However, those strains require long growing cycles due to the low levels of secreted laccase, even no detectable laccase activity. While in strains NITW715076_1 and NITW715076, the laccase activity was found to be 105.83 0.36 U ml 1 and 92.28 0.4 U ml 1 respectively at 50 C . The produced MRGO was extensively attached to IDA-Cu 2+ in our system and exhibited a high capacity for adsorption with CotA laccase. After each purification step, the total protein content and enzyme activity were determined to calculate specific activity and purification factor. The efficacy of dye decolorization depended on pH of the reaction medium. thio used) 2.00 mL (sample size) Title: The optimum thio.) Laccases oxidize a variety of phenolic substrates, performing one-electron oxidations, leading to crosslinking. Breaking barriers: evidencing the ex vivo and in vivo intestinal bypass of intact MOF nanoparticles, further improved via surface functionalization. calculate laccase activity using change in OD @ 420 nm. is sample volume. We don't need to worry about negative signs, so we can just take 27 and divide that by 31.5, and multiply it by 100, and we get 85.7. Hi, The assay for the enzyme laccase is from the following references. (More SS, PS R, Malini S, SM V. Isolation, purification, and characterizatio The decolorization rate calculation formula is shown in Eq. Laccase-like activity was detected in melanin-producing strains of Sinorhizobium meliloti mainly in cells at the stationary growth phase when copper was added to the medium. Molecular weight of the laccase was 63 KDa on SDS-PAGE. 146. Paradoxically, ABTS, the most popular substrate for determining the activity of laccase is not a phenolic compound. In consequence, when ABTS is us Only 1,10-phenanthroline reduced laccase activity under both acidic and alkaline conditions. Dear Christopher Andreas Gasser, Thank you for your post regarding determination of laccase activity.If possible kindly mention the authorof the p Laccase: addressing the ambivalence associated with the calculation of enzyme activity. Now add 0.2 ml of enzyme extract and start recording the change in absorbance for every 30 seconds up to 5 min. with hydrochloric acid because chloride inhibits laccase activity.) What is the function of laccase corresponding gene products? By ABTS method, laccase activity could be detected in all KBS mediums with monoazo, disazo and triphenyl methane dyes (Figure 1). Tukayi Kudanga. The molar absorbances of the substrates were estimated under reaction conditions. Abstract. The laccase activity was normalized to the parental activity at each point, indicating the standard deviation from three independent experiments. Analytical and Bioanalytical Chemistry, 2009. 2a). CAS No. Maximum laccase activity The probable maximum laccase activity may be estimated according to Eq. 4.2. The Effect of pH and Temperature on Purified Laccase. 2,6-Dimethoxyphenol is a phenolic compound that is extensively used for the measurement of laccase activity. Assay of Laccase Activity-Laccase activity was assayed by measurement of oxygen up-take using a Clark oxygen electrode in a vessel (volume: 3 ml), maintained at 25C. For research use only. The effect of different carbon sources on laccase activity by the yeast isolates (Fig. Add 300 ml PEG 6000 solution and stir on a magnetic stirrer for 15 min Laccase activity was determined by the oxidation of ABTS method [ 22 ]. The assay mixture which contained equal volume of the crude enzyme and each buffer at various pH values was incubated at room temperature for 10 minutes, and the determination of laccase activity was done using modified method of 31 Mongkolthanaruk et al (2012). Introduction. So our percent enantiomeric excess is 86%. ABTS and syringaldehyde were effective mediators for decolorization. Sample Laccase Activity = (pmol/minxg) = Units / g or mUnits / mg Where: M = Linear change in oxidized product concentration during t (in pmol) t = t2 - t1 (in min) P = Sample protein content added to well (in g) Unit Definition: One unit of Laccase is the amount of enzyme that produces1 mol of oxidized product per minute at pH 4 at 37C Extracellular laccase or peroxidase activity has been shown to contribute to fungal or oomycete pathogenicity, therefore we further investigate any alteration in such extracellular enzymes activities in the PlMAPK10-silenced transformants, which may account for their reduced pathogenicity. The highest 7.5 CALCULATIONS Units/ml enzyme = A530nm Sample = A530nm/min Test - A530nm/min Blank) (df) (0.001) (0.5) df = dilution factor 0.001 = the change in A530nm/min. 3.4. using the maximum linear rate for both the Test and Blank. Laccases are four-copper oxidases that catalyze the oxidation of phenolic units in lignin as well as a wide range of phenolic compounds and aromatic amines 1.Because of their oxidoreductase activity, laccases can react with 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as a substrate and as a mediator for studying Although the used kits are based on electrochemically active substrates (such as syringaldazine and 3.2. ADVERTISEMENTS: In this article we will discuss about the tests for estimation of polyphenol oxidase enzyme activity. Laccase initial activity as a function of ABTS concentration ([S]) wast to the Michaelis Menten curve, shown ineq 1. The molar extinction coefficient of tetraguaiacol at 470 nm (26.6 10 3 M 1 cm 1) was used for activity calculation 43. Share sensitive information only on official, secure websites. So let's get out the calculator, here. View the article. Price. One unit of laccase activity was defined as the quantity of laccase capable of oxidizing 1 mol of substrate per minute under the assay conditions. Laccase activity is not present in the strain with the SLAC gene disrupted, indicating that SLAC is exported when expressed homologously and that SLAC is the primary extracellular laccase. (2). per unit of laccase at pH 6.5 at 30oC in a 3 mL reaction mix. = 36,000 M-1 cm-1 = 36,000 L mol-1 cm-1, t = 3 min, d = 1 cm, Vt = 3 mL, Vs = 300 L, A = 0.50. Laccase activity was evaluated with different substrates: aniline, DMPD and ABTS (used during the screening of the mutant libraries), and 2,6-dimethoxyphenol -DMP- (a phenolic substrate not targeted during the engineering of the enzyme). One such issue is the question of how the two domains are connected. As the maximum laccase activity of CueO is achieved only in the presence of excess Cu ions 10 1). Localization of Laccase P. martensii NRC 345 was grown on medium No 1 under static and shake culture conditions. Principle: The intensely yellow 2-nitro-5-thio-benzoic acid (TNB) with an absorption maximum at 412 nm reacts with quinones generated through enzymatic oxidation of 4-methyl-catechol (catechol oxidase) and 1, 4 di-hydroxy-benzene (laccase) to yield colourless CotA protein is a component of the endospore coat of Bacillus subtilis and it exhibits the activities of laccase. PFL activity (A) and Laccase activity (B) as a function of enzyme load on the support. However, in certain reactions, direct oxidation is not feasible as laccase can only oxidize The data were processed and analyzed using Origin 8.0 software. A specific amount of laccase enzyme and substrate was added and scanned for a specific time point. Our aim was to minimize its ambivalence, thus, total of five formulas (F1-F5) were used to determine laccase activity of white and blue laccase. = 36,000 M -1 cm -1 = 36,000 L mol -1 cm -1, t = 3 min, d = 1 cm, Vt = 3 mL, Vs = 300 L, A = 0.50. Fourier Transform Infrared Spectroscopy (FTIR) is a fast and versatile technique where spectral evolution profiling, i.e. Cat. Laccase activity calculated using different equations. Characterization analysis. Laccase activity can be determined by using substrates such as ABTS, catechol, Syringaldazine and guaiacol. Determination of laccase activity using The expressed CotA laccase was observed in an active form. The molecular weight of CotA laccase was estimated to be Reports indicate that the activity of this enzyme ranges from 3.5 to 484,000 U L (-1). x 34 (equiv. thio used) 2.00 mL (sample size) Title: Molecular weight of the laccase was 63 KDa on SDS-PAGE. Laccase activity in bacteria was detected for the first time in Azospirillum lipoferum, (QM/MM) calculations were performed to identify which residues are susceptible to oxidation within the active site cavity of the T1 copper; where free radicals are generated after substrate-electron subtraction . (6). Laccase is a key enzyme in the degradation of lignin by fungi. My question is related to Laccase enzyme activity. Do we need to use standard here. As in many research paper about standard is not mentioned. Calculate the Amount of Oxidized Product formed (M). The equation for calculation is shown below: Grams/Liter = 0.10(norm. = [ ] +[ ] V V K S S max m (1) These curves were linearized using the HanesWoolf method, resulting ineq 2. [19]. The activity of laccase against the Cu ions was investigated and it was found that the presence of Cu 2+ influenced significantly the activity of laccase.
laccase activity calculation
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