Yeast Hosts for the Production of Recombinant Laccases: A Review

Abstract

Laccases are multi-copper oxidoreductases which catalyze the oxidation of a wide range of substrates during the simultaneous reduction of oxygen to water. These enzymes, originally found in fungi, plants, and other natural sources, have many industrial and biotechnological applications. They are used in the food, textile, pulp, and paper industries, as well as for bioremediation purposes. Although natural hosts can provide relatively high levels of active laccases after production optimization, heterologous expression can bring, moreover, engineered enzymes with desired properties, such as different substrate specificity or improved stability. Hence, diverse hosts suitable for laccase production are reviewed here, while the greatest emphasis is placed on yeasts which are commonly used for industrial production of various proteins. Different approaches to optimize the laccase expression and activity are also discussed in detail here.

Laccases: Production, Expression Regulation, and Applications in Pharmaceutical Biodegradation

“…The most common heterologous host is the methylotrophic yeast Pichia pastoris with its inducible alcohol oxidase ( AOX1 ) promoter (Antošovǎ and Sychrová, 2016; Ergün and Çalık, 2016). Other hosts, such as prokaryotes (e.g., E. coli and B. subtilis ), yeasts (e.g., Saccharomyces cerevisiae and Yarrowia lipolytica ), filamentous fungi (e.g., Trichoderma reesei and Aspergillus niger ), and plants (e.g., Nicotiana tabacum and O. sativa ), are also used (Piscitelli et al, 2010; Kittl et al, 2012; Liebeton et al, 2014; Mate and Alcalde, 2015; Antošovǎ and Sychrová, 2016).…”

“…Other hosts, such as prokaryotes (e.g., E. coli and B. subtilis ), yeasts (e.g., Saccharomyces cerevisiae and Yarrowia lipolytica ), filamentous fungi (e.g., Trichoderma reesei and Aspergillus niger ), and plants (e.g., Nicotiana tabacum and O. sativa ), are also used (Piscitelli et al, 2010; Kittl et al, 2012; Liebeton et al, 2014; Mate and Alcalde, 2015; Antošovǎ and Sychrová, 2016). In particular, heterologous laccases were constitutively expressed in basidiomycetes Phanerochaete chrysosporium (Coconi-Linares et al, 2015) and C. cinerea (Muraguchi et al, 2011).…”

“…In particular, heterologous laccases were constitutively expressed in basidiomycetes Phanerochaete chrysosporium (Coconi-Linares et al, 2015) and C. cinerea (Muraguchi et al, 2011). Although, filamentous fungi are efficient in protein secretion and have actually given rise to some of the highest recombinant laccase yields reported, genetic techniques are more readily available for yeasts (Piscitelli et al, 2010; Mate and Alcalde, 2015; Antošovǎ and Sychrová, 2016). …”

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

“…Although this enzyme yield is found to be convenient for biochemical characterization, it is still rather low for industrial applications. Production level of CpLcc1 in Pichia pastoris could be further improved by employing several strategies such as the use of native promoters and multiple gene copies, codon optimization, and alteration of secretory signal sequences, as reported previously Antosova and Sychrova, 2016). The tree is calculated with p-distances using Mega ver.…”

“…Among those organisms, yeasts are usually more attractive hosts for heterologous protein production because of their faster microbial growth and ease of gene manipulation, along with the ability to perform posttranslational modifications. In particular, Pichia pastoris, a methylotrophic yeast, has become a very popular host organism for the production of heterologous proteins with a strong inducible alcohol oxidase 1 (AOX1) gene promoter, which results in increased protein bioactivity by reducing proteolytic degradation of the expressed recombinant protein (Macauley-Patrick et al, 2005;Antosova and Sychrova, 2016). White-rot fungi are the best known organisms that can completely degrade lignin to carbon dioxide and water and can further degrade cellulose and hemicelluloses (Eriksson et al, 1990;ten Have and Teunissen, 2001).…”