21.11-14 Journal of pure and applied microbiology Nov. 2014. Vol. 8(Spl. Edn. 2), p. 727-731.pdf (original) (raw)
Related papers
Production and Partial Purification of Laccase from Pseudomonas aeruginosa ADN04
Journal of Pure and Applied Microbiology
The efficient extracellular production of laccase in the liquid culture medium of Pseudomonas aeruginosa ADN04 and the enzyme extraction, characterization and purification were studied. Purified laccase from the culture filtrate of the bacterium has been attained maximum activity after employing ammonium sulphate precipitation, Gel filtration chromatography (Sephadex) and Affinity chromatography DEAE-Sephadex. The molecular mass of the laccase was determined by SDS-PAGE that showed a relative molecular weight of 43 kDa. The enzymatic stability characteristics at different pH and temperature of the purified laccase have been determined. 2,2’-azino-bis (3- ethylbenzothiazoline-6-sulphonic acid) was used as the substrate and have been found to be Vmax is 12.06 and Km is 56.9at 37°C respectively.
IJERT-Production Optimization and Partial Purification of Laccases from Bacterial Consortium
International Journal of Engineering Research and Technology (IJERT), 2014
https://www.ijert.org/production-optimization-and-partial-purification-of-laccases-from-bacterial-consortium https://www.ijert.org/research/production-optimization-and-partial-purification-of-laccases-from-bacterial-consortium-IJERTV3IS060531.pdf Laccase belongs to the family of blue multi-copper oxidases that has three domain structures and usually contains four copper atoms. An attempt was made to screen, optimize, production and partially purify laccase enzyme produced from consortium of laccase producing Pseudomonas aeruginosa and Pseudomonas fluorescens. Guaiacol and acetate buffer were used to assay laccase production. Laccase activity was highest when operated at the following conditions, 72 h incubation, 40°C temperature, and pH-7, 2% Glucose as carbon source and 2% peptone as nitrogen source in the production medium. The enzyme was partially purified by ammonium sulfate precipitation and dialysis method.
Biochemistry Research International, 2015
Laccases are the model enzymes for multicopper oxidases and participate in several applications such as bioremediation, biopulping, textile, and food industries. Laccase producing bacterium,Bacillus subtilisMTCC 2414, was subjected to optimization by conventional techniques and was partially purified using ammonium salt precipitation method. The agroresidue substrates used for higher yield of laccase were rice bran and wheat bran. Maximum production was achieved at temperature 30°C (270 ± 2.78 U/mL), pH 7.0 (345 ± 3.14 U/mL), and 96 h (267 ± 2.64 U/mL) of incubation. The carbon and nitrogen sources resulted in high enzyme yield at 3% sucrose (275 ± 3.11 U/mL) and 3% peptone (352.2 ± 4.32 U/mL) for rice bran and 3% sucrose (247.4 ± 3.51 U/mL) and 3% peptone (328 ± 3.33 U/mL) for wheat bran, respectively. The molecular weights of partially purified laccase were 52 kDa for rice bran and 55 kDa for wheat bran. The laccase exhibited optimal activity at 70°C (260.3 ± 6.15 U/mL), pH 9.0 (2...
Bacterial laccase: recent update on production, properties and industrial applications
3 Biotech, 2017
Laccases (benzenediol: oxygen oxidoreductase, EC 1.10.3.2) are multi-copper enzymes which catalyze the oxidation of a wide range of phenolic and non-phenolic aromatic compounds in the presence or absence of a mediator. Till date, laccases have mostly been isolated from fungi and plants, whereas laccase from bacteria has not been well studied. Bacterial laccases have several unique properties that are not characteristics of fungal laccases such as stability at high temperature and high pH. Bacteria produce these enzymes either extracellularly or intracellularly and their activity is in a wide range of temperature and pH. It has application in pulp biobleaching, bioremediation, textile dye decolorization, pollutant degradation, biosensors, etc. Hence, comprehensive information including sources, production conditions, characterization, cloning and biotechnological applications is needed for the effective understanding and application of these enzymes at the industrial level. The present review provides exhaustive information of bacterial laccases reported till date.
Laccase belongs to the blue multicopper oxidases and participates in cross-linking of monomers, degradation of polymers, and ring cleavage of aromatic compounds. It is widely distributed in higher plants and fungi. It is present in Ascomycetes, Deuteromycetes and Basidiomycetes and abundant in lignin-degrading white-rot fungi. It is also used in the synthesis of organic substance, where typical substrates are amines and phenols, the reaction products are dimers and oligomers derived from the coupling of reactive radical intermediates. In the recent years, these enzymes have gained application in the field of textile, pulp and paper, and food industry. Recently, it is also used in the design of biosensors, biofuel cells, as a medical diagnostics tool and bioremediation agent to clean up herbicides, pesticides and certain explosives in soil. Laccases have received attention of researchers in the last few decades due to their ability to oxidize both phenolic and nonphenolic lignin-related compounds as well as highly recalcitrant environmental pollutants. It has been identified as the principal enzyme associated with cuticular hardening in insects. Two main forms have been found: laccase-1 and laccase-2. This paper reviews the occurrence, mode of action, general properties, production, applications, and immobilization of laccases within different industrial fields.
International Journal of Pharmacy and Pharmaceutical Sciences
Objective: Microorganisms, especially bacteria and its proteins have proven to be potential anti-cancer agents as they selectively attack the tumor cells or tumor micro-environments. The extract of Pseudomonas aeruginosa found to contain proteins that have shown promising anticancer activity. In this work, it was attempted to increase the biomass and trigger the total protein fraction of Pseudomonas aeruginosa (MTCC 2453).Methods: The organism was cultivated in three different such as Luria-Bertani (LB) broth, minimal medium9 (M9), super broth medium (SB) and asparagine-proline (AP) broth. Asparagine proline broth was selected as it has shown high cell growth rate. The media was further optimized by the addition of NaHCO3 and copper sulphate to trigger the protein production. Optimized Aspergine proline broth has achieved highest cell biomass. After the shake flask culture, the overnight grown culture in optimized AP medium was further grown in a 5 L bioreactor by fed-batch cultivat...
Comparison of downstream processing methods in purification of highly active laccase
Bioprocess and Biosystems Engineering
Laccases have received the attention of researchers in the last few decades due to their ability to degrade phenolic and lignin-related compounds. This study aimed at obtaining the highest possible laccase activity and evaluating the methods of its purification. The crude laccase from bioreactor cultivation of Cerrena unicolor fungus was purified using ultrafiltration, aqueous two-phase extraction (ATPE) and foam fractionation (FF), which allowed for the assessment of these three downstream processing (DSP) methods. The repeated fed-batch cultivation mode applied for the enzyme production resulted in a high laccase specific activity in fermentation broth of 204.1 U/mg. The use of a specially constructed spin filter inside the bioreactor enabled the integration of enzyme biosynthesis and biomass filtration in one apparatus. Other methods of laccase concentration and purification, namely ATPE and FF, proved to be useful for laccase separation; however, the efficiency of FF was rather low (recovery yield of 24.9% and purification fold of 1.4). Surprisingly, the recovery yield after ATPE in a PEG 6000-phosphate system in salt phase was higher (97.4%) than after two-step ultrafiltration (73.7%). Furthermore, it was demonstrated that a simple, two-step purification procedure resulted in separation of two laccase isoforms with specific activity of 2349 and 3374 U/mg. All in all, a compact integrated system for the production, concentration and separation of fungal laccases was proposed.
LACCASE: MACRO AND MICROBIAL SOURCES, PRODUCTION, PURIFICATION AND BIOTECHNOLOGICAL APPLICATIONS
Scientific Bulletin. Series F. Biotechnologies, 2019
Laccase belongs to the blue multicopper oxidases and participates in degradation of polymers, ring cleavage of aromatic compounds and cross-linking of monomers. It is distributed in higher plants and fungi. It is present in Ascomycetes, Deuteromycete, and Basidiomycetes and abundant in lignin-degrading white-rot fungi. Lacasse has been reported to be produced by different mushrooms (Trametes, Ganoderma, Pleurotus) and by filamentous bacteria (Streptomyces) or fungi (Aspergillus). The article proposes a comparative analysis of the optimal conditions for laccase production in the case of macromycetes and some micromycetes, like fungi or filamentous bacteria, meanwhile describing the isolation, purification and characterization of the laccase produced by such organisms. All these issues will be approached through the biotechnological application of these enzymes (dye decoloration, bioremediation etc).
PRODUCTION, PARTIAL PURIFICATION AND INDUSRIAL APPLICATION OF LACCASE ENZYME
Human and industrial activities result in the discharge of various pollutants in to the aquatic environment threatening the health of the population and damaging the quality of the environment by rendering water bodies unsuitable. The textile mills daily discharge millions of liters of untreated effluents in the form of wastewater into public drains that eventually empty into rivers. Most of them are recalcitrant in nature, especially azo dyes. In this study, decolourization of dye was carried out by using Laccase enzyme from white rot fungi. At first, white rot fungi was sub cultured on PDA plates by incubating for 5 days. The selected species was inoculated in agro waste substrate for mass production. After incubation for 12 days the enzyme was extracted from the grown culture using sodium acetate buffer. The extracted enzyme was partially purified by using two techniques such as salting out technique and Dialysis. The substrate wheat bran showed more protein concentration 0.373 mg/ml for crude enzyme. Wheat Bran showed more enzyme activity 3.0215 IU/ml after salting out. Protein was separated by SDS-PAGE. The partially purified enzyme was used for textile dye decolourization. According to our work the enzyme laccase produced by white rot fungi effectively decolourize textile dye. Thus, it was found that the dye degradation can be done using laccase enzyme.
A review of microbial laccase production and activity toward different biotechnological applications
Systems Microbiology and Biomanufacturing
Laccases are versatile enzymes that belong to the multi-copper oxidase family. This enzyme has several biotechnological applications because of its ablilty to oxidize a wide range of phenolic and non-phenolic substrates. However, their large-scale applicability in bioremediation and water treatment is hindered by high salt content and extreme pH values of the polluted media which also affects the stability, recovery and recycling of laccase. Apart from some bacteria, laccase is abundantly present in several lignin-degrading white-rot fungi viz. Ascomycetes, Deuteromycetes, and Basidiomycetes. Recently, laccase has been employed in the development of biosensors as a medical diagnostic tool, biofuel cells, and in bioremediation purpose to remove herbicides, pesticides, and some toxic chemicals from the soil. However, most of the enzymes including laccase are normally unstable and susceptible to lose their activity over time. This might be avoided by maintaining the activity and lengthening the enzyme's lifespan through the use of appropriate immobilization procedures. The potential of laccase immobilized biocathodes for dye decolorization in microbial fuel cells has recently been studied. Immobilized laccase nanoparticles have potential uses as biocatalyst in the bioremediation of pollutants. In addition, advanced research considering microbial laccase has been conducted for its heterologous expression along with in silico protein engineering to attain maximum enzyme activity which can be potentially applied in different biotechnological sectors. Patent related to laccase also implied that this enzyme can be used as suitable catalyst for the production of promising anti-cancer drugs and even as a significant ingredient in cosmetics.