title: Expression of a new laccase from Moniliophthora roreri at high levels in Pichia pastoris and its potential application in micropollutant degradation.
abstract: Laccases have gained significant attention due to their emerging applications including bioremediation, biomass degradation and biofuel cells. One of the prerequisites for the industrial application of laccases is their sufficient availability. However, expression levels of recombinantly expressed laccases are often low. In this study Mrl2, a new laccase from the basidiomycete Moniliophthora roreri, was cloned in Pichia pastoris and produced in an optimized fed-batch process at an exceptionally high yield of 1.05 g l-1. With a redox potential of 0.58 V, Mrl2 belongs to mid-redox potential laccases. However, Mrl2 demonstrated high kcatvalues of 316, 20, 74, and 36 s-1towards 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), syringaldazine (SGZ), 2,6-dimethoxyphenol (2,6-DMP) and guaiacol, respectively. Mrl2 remained stable above pH 6 and in the presence of many metal ions, which is important for application in bioremediation. Mrl2 was investigated for the ability to degrade endocrine disrupting chemicals (EDCs) and non-steroidal anti-inflammatory drugs (NSDAIs) at neutral pH value. The enzyme accepted and converted estrone, 17β-estradiol, estriol, the synthetic contraceptive 17α-ethinyl estradiol and bisphenol A at pH 7 faster than high-potential laccases from Trametes versicolor. For example, within 30 min Mrl2 removed more than 90% bisphenol A, 17ß-estradiol, 17α-ethinyl estradiol and estriol, respectively. The concentration of the recalcitrant drug diclofenac dropped by 56% after 20 h incubation with Mrl2.
authors.affiliation: Institute of Biochemistry II, Heinrich-Heine University Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.|Bioeconomy Science Center (BioSC), Heinrich-Heine University Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.
authors.forename: Agathe
authors.initials: A
authors.lastname: Bronikowski
authors.affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC, Delft, The Netherlands.
authors.forename: Peter-Leon
authors.initials: PL
authors.lastname: Hagedoorn
authors.affiliation: Institute of Biochemistry II, Heinrich-Heine University Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.|Bioeconomy Science Center (BioSC), Heinrich-Heine University Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Koschorreck
authors.affiliation: Institute of Biochemistry II, Heinrich-Heine University Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany. vlada.urlacher@uni-duesseldorf.de.|Bioeconomy Science Center (BioSC), Heinrich-Heine University Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany. vlada.urlacher@uni-duesseldorf.de.
authors.forename: Vlada B
authors.initials: VB
authors.lastname: Urlacher
keywords: Expression; Laccase; Micropollutant degradation; Pichia pastoris