High-yield production of manganese peroxidase, lignin peroxidase, and versatile peroxidase in Phanerochaete chrysosporium

Abstract

The white-rot fungus Phanerochaete chrysosporium secretes extracellular oxidative enzymes during secondary metabolism, but lacks versatile peroxidase, an enzyme important in ligninolysis and diverse biotechnology processes. In this study, we report the genetic modification of a P. chrysosporium strain capable of co-expressing two endogenous genes constitutively, manganese peroxidase (mnp1) and lignin peroxidase (lipH8), and the codon-optimized vpl2 gene from Pleurotus eryngii. For this purpose, we employed a highly efficient transformation method based on the use of shock waves developed by our group. The expression of recombinant genes was verified by PCR, Southern blot, quantitative real-time PCR (qRT-PCR), and assays of enzymatic activity. The production yield of ligninolytic enzymes was up to four times higher in comparison to previously published reports. These results may represent significant progress toward the stable production of ligninolytic enzymes and the development of an effective fungal strain with promising biotechnological applications.

Progress and obstacles in the production and application of recombinant lignin-degrading peroxidases

“…42 Recently, native LiP and MnP were expressed in P. chrysosporium in concert with an exogenous VP, generating a recombinant strain with higher LiP and MnP activity and detectable VP activity. 43 Further strain engineering included the introduction of an additional laccase. 44 In addition to LiP, MnP and VP, fungal DyP-type peroxidases have also been expressed in various heterologous production platforms, [45][46][47][48] with yields of 0.1 mg L ¡1 purified enzyme achieved using E. coli 47 and up to 62 mg L ¡1 recovered from the supernatant of A. oryzae expression cultures.…”

A valuable peroxidase activity from the novel species Nonomuraea gerenzanensis growing on alkali lignin

“…In fact, when compared to P. chrysosporium MnP, our N. gerenzanensis peroxidase preparation shows a significantly higher stability at pH >6.5 and a higher thermostability ( P. chrysosporium MnP is fully inactivated in ≈ 3 min at 55 °C) [46], [48]. Although more active at acidic than at basic pHs, N. gerenzanensis peroxidase activity is more stable at higher pHs than the fungal counterparts.…”

Ligninolytic enzymes: Versatile biocatalysts for the elimination of endocrine‐disrupting chemicals in wastewater

“…It is also referred to as “hybrid peroxidase” or “lignin‐manganese peroxidase” and is largely produced by ligninolytic fungi belonging to certain genera: Bjerkandera (Heinfling, Martınez, Martınez, Bergbauer, & Szewzyk, ), Pleurotus (Palma, Lloret, Sepulveda, & Contreras, ; Ruiz‐Duenas, Martınez, & Martınez, ), and Lepista (Zorn, Langhoff, Scheibner, Nimtz, & Berger, ). Its production by Phanerochaete chrysosporium has also been reported (Coconi‐Linares et al., ). Nonetheless, there is limited information on its production by bacteria.…”

“…The resultant complex will in turn oxidize the phenolic component of lignin structure which leads to generation of unstable radicals that may breakdown naturally (Hofrichter, 2002 Bjerkandera (Heinfling, Martınez, Martınez, Bergbauer, & Szewzyk, 1998), Pleurotus (Palma, Lloret, Sepulveda, & Contreras, 2016;Ruiz-Duenas, Martınez, & Martınez, 1999), and Lepista (Zorn, Langhoff, Scheibner, Nimtz, & Berger, 2003). Its production by Phanerochaete chrysosporium has also been reported (Coconi-Linares et al, 2014).…”