A Highly Efficient Recombinant Laccase from the Yeast Yarrowia lipolytica and Its Application in the Hydrolysis of Biomass

Abstract

A modified thermal asymmetric interlaced polymerase chain reaction was performed to obtain the first yeast laccase gene (YlLac) from the isolated yeast Yarrowia lipolytica. The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris. YlLac is a monomeric glycoprotein with a molecular mass of ~55 kDa as determined by polyacrylamide-gel electrophoresis. It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s-1 μM-1) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s-1 μM-1) than other reported laccases. The standard redox potential of the T1 site of the enzyme was found to be 772 mV. The highest catalytic efficiency of the yeast recombinant laccase, YlLac, makes it a good candidate for industrial applications: it removes phenolic compounds in acid-pretreated woody biomass (Populus balsamifera) and enhanced saccharification.

Heterologous expression and characterization of a high redox potential laccase from Coriolopsis polyzona MUCL 38443

“…Recombinant CpLcc1 laccase was strongly inhibited by 1 mM sodium azide and 1 mM SDS by 90.12% and 84.90%, respectively, while 0.1 mM sodium azide and 1 mM L-cysteine also caused 55.62% and 25.34% inhibition, respectively (Table 5). These results were in accordance with the results from previous reports (Garcia et al, 2007;More et al, 2011;Kalyani et al, 2015).…”

“…Thus, in the present study, effect of temperature on the enzyme activity was analyzed against ABTS between 20 °C and 90 °C. Consistent with a previous report (Kalyani et al, 2015), the optimum temperature of the recombinant enzyme CpLcc1 was found as 70 °C (Figure 6a). The half-life of its activity (t 1/2 ) was longer than 8 h at both 30 °C and 40 °C and it was 30 min at 50 °C (Figure 6b).…”

Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge

“….2) is a blue copper oxidase with a broad range of substrates (Kalyani et al 2015), varying from organic to inorganic compounds such as diphenols, aromatic amines, polyphenols (Hoegger et al 2006;Rivera-Hoyos et al 2013;Sharma et al 2007), and Mn 2+ (Brouwers et al 1999). The reactions of laccase are mediated by a one-electron transfer mechanism (Thurston 1994).…”

Bio-Prospecting Laccases in the Bacterial Diversity of Activated Sludge From Pulp and Paper Industry

“…1 g of activated sludge was mixed with 9 ml of normal saline and was shaken for 1 h with glass beads on a horizontal shaker. This suspension was centrifuged and supernatant was used for assaying the activity using 2 mM ABTS at different pH (5,6,7,8) and temperatures 37-75°C. Controls were set up using autoclaved sludge suspension.…”

“…Laccase is a multicopper enzyme found in a wide range of taxa including plants, fungi, bacteria, insects and bacteria. Recently, laccase was also reported in yeast [8]. It performs reactions in ecofriendly manner, requires only oxygen to catalyze the oxidation of phenolic compounds and produces water as by product.…”