Improvement of Laccase Production by Thielavia terrestris Co3Bag1. Enhancing the Bio-Catalytic Performance of the Native Thermophilic TtLacA via Immobilization in Copper Alginate Gel Beads

Abstract

A 32-fold increase in laccase activity production by the thermophilic biomass-degrading fungus T. terrestris Co3Bag1 was achieved when the microorganism was grown on a modified medium containing fructose, sodium nitrate, and copper. A 70 kDa laccase (TtLacA), produced under the above conditions, was purified, immobilized in copper alginate gel beads, and characterized. TtLacA, both free and immobilized enzymes, exhibited optimal activity at pH 3.0, at a temperature of 65 and 70 °C, respectively, although both displayed 70% of activity from 40 to 70 °C. Free and immobilized enzymes retained at least 80% of relative activity in the pH range from 3 to 4.6. Immobilized TtLacA manifested a 2.3-fold higher thermal stability than the free form of the enzyme at 60 and 70 °C. Immobilized TtLacA retained 95% initial activity for six consecutive reuse cycles at 60 °C, and also retained 86% of initial activity after 12 days of storage at 4 °C. Based on the biochemical features, thermophilic TtLacA may be an efficient enzyme for dye decolorization and other industrial applications at high temperatures or acidic conditions. This work represents the first report about the immobilization and biochemical characterization of a thermophilic laccase from a member of the genus Thielavia.

Purification, characterization and three-dimensional structure prediction of multicopper oxidase Laccases from Trichoderma lixii FLU1 and Talaromyces pinophilus FLU12

“…Tl FLU1 might have a more robust cold-shock response mechanism that allows it to maintain higher Laccase activity at lower temperatures compared to Tp FLU12. This aligns with a previous report, where Laccase from Thielavia terrastris Co3Bag1 retains 86% of its activity after 12 d storage at 4 ˚C due to cold-shock response 57 . Similarly, it has been reported that Laccases from fungi can retain activity at low temperature (4–7 °C) 58 .…”

“…Conversely, the substantial loss of activity in Tp FLU12 below 20 °C after 24 h of incubation could be since Laccase like many enzymes, has an optimal temperature range for activity 59 . At higher temperatures, fungi Laccases have been reported to possess characteristics of retaining more activity above 20 °C after 24 h 57 . Also, Psychrophilic Laccases, such as those produced by Kabatiella bupleuri G3 IBMiP which have been documented to have exhibit an optimal temperature range of 30–40 °C, with a significant decrease in activity below this ranges but still retains some activity even at 10 °C.…”

Kinetic studies on optimized extracellular laccase from Trichoderma harzianum PP389612 and its capabilities for azo dye removal

“…The laccase enzyme feature (EC 1.10.3.2) belongs to multicopper polyphenol oxidase; substrate specificity increases through a redox mediator's addition to the reaction mixture [1,2]. It works as a catalyst because of the copper atoms in the enzyme's structure: blue paramagnetic copper, non-blue paramagnetic copper, and two spin-coupled copper pairs [3]. Its stability is remarkable due to the carbohydrate structure portion, which protects the enzyme from inactivation by free radicals and proteolysis [4].…”