Limits...
A novel multicopper oxidase (laccase) from cyanobacteria: Purification, characterization with potential in the decolorization of anthraquinonic dye

View Article: PubMed Central - PubMed

ABSTRACT

A novel extracellular laccase enzyme produced from Spirulina platensis CFTRI was purified by ultrafiltration, cold acetone precipitation, anion exchange and size exclusion chromatography with 51.5% recovery and 5.8 purification fold. The purified laccase was a monomeric protein with molecular mass of ~66 kDa that was confirmed by zymogram analysis and peptide mass fingerprinting. The optimum pH and temperature of the enzyme activity was found at 3.0 and 30°C using ABTS as substrate but the enzyme was quite stable at high temperature and alkaline pH. The laccase activity was enhanced by Cu+2, Zn+2 and Mn+2. In addition, the dye decolorization potential of purified laccase was much higher in terms of extent as well as time. The purified laccase decolorized (96%) of anthraquinonic dye Reactive blue- 4 within 4 h and its biodegradation studies was monitored by UV visible spectra, FTIR and HPLC which concluded that cyanobacterial laccase can be efficiently used to decolorize synthetic dye and help in waste water treatment.

No MeSH data available.


Effect of inhibitors on laccase activity was measured using ABTS as a substrate.Values are mean of triplicates ± S.E
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5383238&req=5

pone.0175144.g005: Effect of inhibitors on laccase activity was measured using ABTS as a substrate.Values are mean of triplicates ± S.E

Mentions: The purified laccase enzyme had been studied presence of studied inhibitors (1mM). The enzyme was inhibited by all studied inhibitors (Fig 5). The order of inhibition was sodium azide (98.23%) > L-cysteine (94.61%) > Thioglycollic acid (82.41%) > Thiourea (60.23%) > EDTA (30.64%). Other scientists have also reported the inhibition of laccase activity by sodium azide Mannaporthe grisea [46] and Pleurotus ostreatus [25]. Inhibition of laccase activity by thioglycollic acid was reported in Chaetomium thermophilium which may be due to effect of copper at the catalytic centre of laccase [47]. It was reported that L-cysteine is one of the effective inhibitor for fungal laccase and like thioglycollic acid, due it effect of copper at the catalytical centre [48]. Many scientistist have reported that EDTA is also an inhibitor of metallo enzymes due to its tendency of forming inactive complexes with inorganic prosthetic cofactors of the enzyme [49]. The laccase activity was inhibited by EDTA was also reported by many researchers [43, 50].


A novel multicopper oxidase (laccase) from cyanobacteria: Purification, characterization with potential in the decolorization of anthraquinonic dye
Effect of inhibitors on laccase activity was measured using ABTS as a substrate.Values are mean of triplicates ± S.E
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383238&req=5

pone.0175144.g005: Effect of inhibitors on laccase activity was measured using ABTS as a substrate.Values are mean of triplicates ± S.E
Mentions: The purified laccase enzyme had been studied presence of studied inhibitors (1mM). The enzyme was inhibited by all studied inhibitors (Fig 5). The order of inhibition was sodium azide (98.23%) > L-cysteine (94.61%) > Thioglycollic acid (82.41%) > Thiourea (60.23%) > EDTA (30.64%). Other scientists have also reported the inhibition of laccase activity by sodium azide Mannaporthe grisea [46] and Pleurotus ostreatus [25]. Inhibition of laccase activity by thioglycollic acid was reported in Chaetomium thermophilium which may be due to effect of copper at the catalytic centre of laccase [47]. It was reported that L-cysteine is one of the effective inhibitor for fungal laccase and like thioglycollic acid, due it effect of copper at the catalytical centre [48]. Many scientistist have reported that EDTA is also an inhibitor of metallo enzymes due to its tendency of forming inactive complexes with inorganic prosthetic cofactors of the enzyme [49]. The laccase activity was inhibited by EDTA was also reported by many researchers [43, 50].

View Article: PubMed Central - PubMed

ABSTRACT

A novel extracellular laccase enzyme produced from Spirulina platensis CFTRI was purified by ultrafiltration, cold acetone precipitation, anion exchange and size exclusion chromatography with 51.5% recovery and 5.8 purification fold. The purified laccase was a monomeric protein with molecular mass of ~66 kDa that was confirmed by zymogram analysis and peptide mass fingerprinting. The optimum pH and temperature of the enzyme activity was found at 3.0 and 30°C using ABTS as substrate but the enzyme was quite stable at high temperature and alkaline pH. The laccase activity was enhanced by Cu+2, Zn+2 and Mn+2. In addition, the dye decolorization potential of purified laccase was much higher in terms of extent as well as time. The purified laccase decolorized (96%) of anthraquinonic dye Reactive blue- 4 within 4 h and its biodegradation studies was monitored by UV visible spectra, FTIR and HPLC which concluded that cyanobacterial laccase can be efficiently used to decolorize synthetic dye and help in waste water treatment.

No MeSH data available.