Microbial enzyme systems for lignin degradation and their transcriptional regulation

Lignocellulosic biomass is the most abundant renewable resource in nature and has received considerable attention as one of the most promising alternatives to oil resources for the provision of energy and certain raw materials. The phenolic polymer lignin is the second most abundant constituent of this biomass resource and has been shown to have the potential to be converted into industrially important aromatic chemicals after degradation. However, due to its chemical and structural nature, it exhibits high resistance toward mechanical, chemical, and biological degradation, and this causes a major obstacle for achieving efficient conversion of lignocellulosic biomass. In nature, lignin-degrading microorganisms have evolved unique extracellular enzyme systems to decompose lignin using radical mediated oxidative reactions. These microorganisms produce a set of different combinations of enzymes with multiple isozymes and isoforms by responding to various environmental stimuli such as nutrient availability, oxygen concentration and temperature, which are thought to enable effective decomposition of the lignin in iignocellulosic biomass. In this review, we present an overview of the microbial ligninolytic enzyme systems including general molecular aspects, structural features, and systematic differences in each microorganism. We also describe the gene expression pattern and the transcriptional regulation mechanisms of each ligninolytic enzyme with current data.

Transcriptome exploration to provide a resource for the study of Auricularia heimuer

Auricularia heimuer,an edible jelly fungus,is in considerable demand in Asia due to its high nutritive,economic and medicinal values.RNA-Seq was used to investigate and analyze the mycelium transcriptome of A.heimuer for gene discovery.A total of 26,857 unigenes with an N50 length of 1333 bp were assembled by de novo sequencing.In addition,unigenes were annotated by publicly available databases,including gene descriptions,gene ontology(GO),clusters of orthologous group(COG),Kyoto Encyclopedia of Genes and Genomes(KEGG)metabolic pathways,and protein family(Pfam)terms.A.heimuer was also studied for its wood degradation ability.Thirty-eight putative FOLymes(fungal oxidative lignin enzymes)and 251 CAZymes(carbohydrate-active enzymes)were located from A.heimuer transcriptome.Our study provides a comprehensive sequence resource for A.heimuer at the transcriptional level,which will lay a strong foundation for functional genomics studies and gene discovery of this promising fungus.

The effect of particle size and amount of inoculum on fungal treatment of wheat straw and wood chips

Background： The aim of this study was to optimize the fungal treatment of lignocellulosic biomass by stimulating the colonization. Wheat straw and wood chips were treated with Ceriporiopsis subvermispora and Lentinula edodes with various amounts of colonized millet grains（0.5, 1.5 or 3.0 % per g of wet weight of substrate） added to the substrates. Also, wheat straw and wood chips were chopped to either 0.5 or 2 cm.Effectiveness of the fungal treatment after 0, 2, 4, 6, or 8 wk of incubation was determined by changes in chemical composition, in vitro gas production（IVGP） as a measure for rumen degradability, and ergosterol content as a measure of fungal biomass.Results： Incomplete colonization was observed for C. subvermispora treated wheat straw and L. edodes treated wood chips. The different particle sizes and amounts of inoculum tested, had no significant effects on the chemical composition and the IVGP of C. subvermispora treated wood chips. Particle size did influence L.edodes treatment of wheat straw. The L. edodes treatment of 2 cm wheat straw resulted in a more selective delignification and a higher IVGP than the smaller particles. Addition of 1.5 % or 3 % L. edodes inoculum to wheat straw resulted in more selective delignification and a higher IVGP than addition of 0.5 % inoculum.Conclusion： Particle size and amount of inoculum did not have an effect on C. subvermispora treatment of wood chips. At least 1.5 % L. edodes colonized millet grains should be added to 2 cm wheat straw to result in an increased IVGP and acid detergent lignin（ADL） degradation.

Laccases from Actinobacteria—What We Have and What to Expect

Laccases are blue multicopper enzymes, capable of oxidizing diverse aromatic and non-aromatic compounds of industrial interest, concomitantly with reduction of molecular oxygen to water. Tolerance to extreme conditions, such as high temperature, salinity or extreme pH, is required for practical industrial applications. Here we focus on bacterial laccases from the phylum Actinobacteria, notably the order Actinomycetales. Currently, less than 10 enzymes have been properly characterized, all belonging to genus Streptomyces, but it is noteworthy that all of them have exhibited industrially important properties. Furthermore, studies with enzymes from this phylum revealed a novel molecular structure of laccases, providing the basis for a distinct family, the two-domain laccases. The relevant traits of actinomycetes laccases emphasize the need for more studies involving the isolation of this bacterial group from lignin-rich environmental samples, detection of their laccase activity and thereafter, characterization of the proteins and related genes. The nonhomogeneous responses of actinomycetes laccases to traditional inhibitors, substrates or metal ions have challenged the currently accepted “laccase concept”. Finally, considering that distinguishing laccase activity in vitro from other ligninolytic enzymes becomes a difficult task due to overlaps in catalytical properties of the enzymes, we proposed a simple flow chart to help experimental assays.