| Size | Price | Stock | Qty |
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| 500mg |
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| 1g |
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| Other Sizes |
| Targets |
Laccase targets phenolic and aromatic compounds as its substrates. The enzyme catalyzes the one-electron oxidation of these substrates, generating reactive radicals that can undergo further reactions such as polymerization, depolymerization, or cross-linking. The enzyme's active site contains multiple copper ions that are involved in the electron transfer process. Laccase reduces molecular oxygen to water, accepting four electrons from four substrate molecules. The enzyme has low substrate specificity, acting on both o- and p-quinols, aminophenols, and phenylenediamine. In lignin degradation, laccase targets the phenolic subunits of lignin, generating radicals that lead to the breakdown of the lignin polymer. In cell wall synthesis, laccase promotes the oxidative coupling of monolignin to form lignin. The enzyme's antimicrobial activity is thought to be mediated by the generation of reactive oxygen species and the oxidation of microbial cell wall components. The enzyme's cytotoxicity against cancer cells may be due to the generation of reactive oxygen species and the induction of oxidative stress.
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| ln Vitro |
In vitro, laccase exhibits antimicrobial activity and cytotoxicity that can be used in cancer research. The enzyme catalyzes the oxidation of various phenolic and aromatic compounds, which can be measured spectrophotometrically. For example, laccase activity is commonly measured using substrates such as ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), syringaldazine, or pyrocatechol. The oxidation of these substrates produces colored products that can be measured at specific wavelengths. In antimicrobial assays, laccase is tested against various bacterial and fungal strains to assess its ability to inhibit microbial growth. In cancer research, laccase is tested against cancer cell lines to assess its cytotoxicity. The enzyme's ability to degrade lignin and other phenolic compounds is studied in in vitro assays using lignin model compounds or natural lignin substrates. The enzyme's activity is influenced by pH, temperature, and the presence of mediators such as ABTS or syringaldazine.
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| ln Vivo |
In vivo, laccase plays a role in lignin degradation, cell wall synthesis, and has potential in food polymer cross-linking. The enzyme is produced by various microorganisms, especially fungi and bacteria, and is involved in the degradation of plant cell wall components. In fungi, laccase is involved in the degradation of lignin, which is a major component of wood, and is important for the recycling of carbon in ecosystems. The enzyme is also involved in the synthesis of cell wall components, including the oxidative coupling of monolignin to form lignin. In industrial applications, laccase is used in bioremediation to degrade environmental pollutants, in pulp and paper processing to delignify wood pulp, in textile dye decolorization, and in food processing to cross-link food polymers. The enzyme's in vivo applications are primarily industrial rather than therapeutic. However, its antimicrobial and cytotoxic activities suggest potential therapeutic applications, although these have not been extensively explored in vivo.
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| Enzyme Assay |
In vitro enzyme assays for laccase typically involve the use of chromogenic or fluorogenic substrates. A common substrate is ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), which is oxidized by laccase to a green-colored radical cation that absorbs at 420 nm. Another common substrate is syringaldazine, which is oxidized to a purple-colored product that absorbs at 530 nm. Pyrocatechol is also used as a substrate, with the oxidation product measured at 400 nm. A typical assay protocol involves incubating laccase (0.01-0.1 U/mL) with the substrate (0.1-10 mM) in an appropriate buffer (e.g., sodium acetate buffer, pH 4.5-5.5) at 25-37°C. The increase in absorbance is monitored over time, and the enzyme activity is calculated from the initial linear rate of the reaction. One unit (U) corresponds to the amount of enzyme which converts 1 μmol of pyrocatechol per minute at pH 4.5 and 25°C. For kinetic characterization, assays are performed at various substrate concentrations, and kinetic parameters (Km, Vmax) are determined from Lineweaver-Burk plots. Inhibition studies are conducted by pre-incubating the enzyme with potential inhibitors before adding the substrate.
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| Cell Assay |
In vitro cell-based assays for laccase are performed using cancer cell lines to study its cytotoxicity. Cells are cultured in appropriate medium and treated with laccase at various concentrations (typically 0.1-10 U/mL) for 24-72 hours. Following treatment, cell viability is assessed using MTT, CCK-8, or LDH assays. Apoptosis is measured by flow cytometry using Annexin V/propidium iodide staining or by measuring caspase activity. The generation of reactive oxygen species (ROS) is measured using fluorescent probes such as DCFH-DA. For antimicrobial assays, bacterial or fungal cultures are grown in the presence of laccase, and microbial growth is monitored by measuring optical density or by colony counting. Each experiment includes appropriate controls (untreated cells, vehicle controls) and is performed in triplicate to ensure statistical reliability. The enzyme is typically dissolved in buffer (e.g., sodium acetate buffer, pH 4.5-5.5) for use in these assays.
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| Animal Protocol |
In vivo animal experiments with laccase are limited, as the enzyme is primarily used in industrial and research applications rather than as a therapeutic agent. However, studies have been conducted to investigate the enzyme's potential in bioremediation and other environmental applications. In these studies, laccase is typically applied to contaminated soil or water, and the degradation of pollutants is measured over time. In some cases, laccase has been encapsulated or immobilized to improve its stability and reusability. The enzyme's potential therapeutic applications, such as antimicrobial or anticancer therapy, have not been extensively explored in vivo. Further in vivo studies are needed to fully characterize the enzyme's pharmacokinetic and pharmacodynamic properties for therapeutic applications.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of laccase are not applicable, as the enzyme is used as a research and industrial tool rather than as a therapeutic agent. Laccase is a protein enzyme that would be degraded if administered orally and would likely be immunogenic if administered parenterally. In industrial applications, the enzyme is typically used in immobilized form to improve its stability and reusability. The enzyme's activity is influenced by pH, temperature, and the presence of mediators. Laccase is most active at acidic pH (pH 4.5-5.5) and at moderate temperatures (25-50°C). The enzyme's stability can be improved by immobilization or by the addition of stabilizers. As with all enzymes, the pharmacokinetics of laccase would be characterized by rapid clearance and potential immunogenicity if administered in vivo.
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| Toxicity/Toxicokinetics |
The toxicological profile of laccase is generally favorable, as the enzyme is widely used in industrial applications and is considered safe for these purposes. The enzyme is produced by various microorganisms, especially fungi and bacteria, and is naturally present in the environment. However, the enzyme's potential toxicity in humans has not been extensively studied. In cell-based assays, laccase has been shown to have cytotoxic activity against cancer cells, suggesting that it may have potential as an anticancer agent. However, this cytotoxicity may also pose a risk to normal cells. The enzyme's immunogenicity is a potential concern if it were to be used as a therapeutic agent. The enzyme is classified as a research chemical and is not approved for human use. Standard safety precautions should be observed when handling the enzyme, including the use of appropriate personal protective equipment. As with all enzymes, inhalation and skin contact should be avoided.
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| References |
[1]. Grzegorz Janusz, et al. Laccase Properties, Physiological Functions, and Evolution. Int J Mol Sci. 2020 Jan 31;21(3):966.
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| Additional Infomation |
A copper-containing oxidoreductase catalyzes the oxidation of 4-benzenediol to 4-benzenesymquinone. It is also active against various ortho- and para-quinones. This enzyme is primarily found in fungi and participates in lignin degradation, pigment biosynthesis, and the detoxification of lignin derivatives.
Laccase (Denilite IIS) is a valuable research and industrial enzyme with applications in bioremediation, pulp and paper processing, textile dye decolorization, and food processing. It is a multicopper oxidase widely found in microorganisms, plants, and fungi that catalyzes the one-electron oxidation of various phenolic and aromatic compounds. The enzyme is a group of multi-copper proteins of low specificity acting on both o- and p-quinols, and often acting also on aminophenols and phenylenediamine. Laccase is involved in lignin degradation, cell wall synthesis, and has potential in food polymer cross-linking. It also exhibits antimicrobial activity and cytotoxicity that can be used in cancer research. One unit (U) corresponds to the amount of enzyme which converts 1 μmol of pyrocatechol per minute at pH 4.5 and 25°C. The enzyme is not approved for any clinical indication and is strictly for research and industrial use only. Its versatility and broad substrate specificity make it a valuable tool for various biotechnological applications. |
| Molecular Formula |
C9H13NO
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|---|---|
| Molecular Weight |
151.205622434616
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| Exact Mass |
1071.472
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| CAS # |
80498-15-3
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| PubChem CID |
3153309
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| Appearance |
Off-white to light yellow solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
1611.6±65.0 °C at 760 mmHg
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| Flash Point |
928.5±34.3 °C
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| Vapour Pressure |
0.0±0.6 mmHg at 25°C
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| Index of Refraction |
1.566
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| LogP |
-1.33
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
1
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
11
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| Complexity |
147
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=CC1C=C(C)N(CC)C=1C
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| InChi Key |
NWDZDFOKSUDVJV-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C9H13NO/c1-4-10-7(2)5-9(6-11)8(10)3/h5-6H,4H2,1-3H3
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| Chemical Name |
1-ethyl-2,5-dimethylpyrrole-3-carbaldehyde
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 6.6133 mL | 33.0666 mL | 66.1332 mL | |
| 5 mM | 1.3227 mL | 6.6133 mL | 13.2266 mL | |
| 10 mM | 0.6613 mL | 3.3067 mL | 6.6133 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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