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Investigations on the Fusants From Wide Cross Between White-Rot Fungi and Saccharomyces cerevisiae Reveal Unknown Lignin Degradation Mechanism.

ABSTRACT: The degradation of lignocellulose by fungi, especially white-rot fungi, contributes a lot to carbon cycle, bio-fuel production, and many other bio-based applications. However, the existing enzymatic and non-enzymatic degradation mechanisms cannot be unequivocally supported by in vitro simulation experiment, meaning that additional mechanisms might exist. Right now, it is still very difficult to discover new mechanisms with traditional forward genetic approaches. To disclose novel lignin degradation mechanisms in white-rot fungi, a series of fusants from wide cross by protoplast fusion between Pleurotus ostreatus, a well-known lignin-degrading fungus, and Saccharomyces cerevisiae, a well-known model organism unable to degrade lignocellulose, was investigated regarding their abilities to degrade lignin. By analyzing the activity of traditional lignin-degrading enzyme, the ability to utilize pure lignin compounds and degrade corn stalk, a fusant D1-P was screened out and proved not to contain well-recognized lignin-degrading enzyme genes by whole-genome sequencing. Further investigation with two-dimension nuclear magnetic resonance (NMR) shows that D1-P was found to be able to degrade the main lignin structure β-O-4 linkage, leading to reduced level of this structure like that of the wild-type strain P. ostreatus after a 30-day semi-solid fermentation. It was also found that D1-P shows a degradation preference to β-O-4 linkage in A_β(S)-threo. Therefore, wide cross between white-rot fungi and S. cerevisiae provides a powerful tool to uncover novel lignocellulose degradation mechanism that will contribute to green utilization of lignocellulose to produce bio-fuel and related bio-based refinery.

SUBMITTER: Shao Q

PROVIDER: S-EPMC9310788 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Indigenous white-rot fungal isolates Schizophyllum commune, Phanerochaete chrysosporium, Ganoderma racenaceum, and Lentinus squarrosulus, demonstrating the ability to depolymerize lignin of the crop residues, were studied for their potential to produce ligninolytic enzymes using modified production media under conditions of limiting and excess nitrogen for higher enzymatic expressions. Secretome-rich media on the investigation confirmed the successful production of lignin-depolymerizing enzymes, viz. laccase, lignin peroxidase, manganese peroxidase, and versatile peroxidase. Production of laccases and peroxidases was statistically significant in nitrogen-limiting media with and without the substrate, across all white-rot fungal cultures at 95% confidence interval. Nitrogen-limiting media with the substrate on analysis extracellularly expressed 99.27 U of laccase and 68.48 U of manganese peroxidase in Schizophyllum commune, while 195.14 U of lignin peroxidase was produced by Phanerochaete chrysosporium. Lentinus squarrosulus expressed 455.34 U of laccase and 357.13 U of versatile peroxidase with 250.09 U of laccase and 206.95 U of manganese peroxidase produced by Ganoderma racenaceum for every milliliter of the media used. Nitrogen-limiting media triggered the production of laccase during the initial stages of growth while the expression of peroxidases was predominant at a later stage. Also, this media evinced increased enzymatic yields with low biomass content compared to nitrogen-excess conditions. The extant study confirmed the positive influence of nitrogen-limiting media in the efficient production of ligninolytic enzymes and their suggestive degradation potential for environmental pollutants, making these enzymes a safe, clean alternative to the use of chemicals and the media to be effective for large-scale production of ligninolytic enzymes. IMPORTANCE Lignin on account of its high abundance, complex polymeric structure, and biochemical properties is identified as a promising candidate in renewable energy and bioproduct manufacturing. However, depolymerization of lignin remains a major challenge in lignin utilization, entailing the employment of harsh treatments representing not only an environmental concern but also a waste of economic potential. Developing an alternative green technology to minimize this impact is imperative. Methods using enzymes to depolymerize lignin are the focus of recent studies. Current research work emphasized the efficient expression of the major lignin-depolymerizing enzymes: laccases, lignin peroxidases, manganese peroxidases, and versatile peroxidases from native isolates of white-rot fungus for several biotechnological applications as well as treatment of crop residues for use as ruminant feed in improving productivity. The importance of nitrogen in augmenting the expression of lignin-depolymerizing enzymes and providing a media recipe for the cost-effective production of ligninolytic enzymes is highlighted.

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Investigations on the Fusants From Wide Cross Between White-Rot Fungi and Saccharomyces cerevisiae Reveal Unknown Lignin Degradation Mechanism.

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