BackgroundThe filamentous fungus Penicillium oxalicum is a potential alternative to Trichoderma reesei for industrial production of a complete cellulolytic enzyme system for a bio-refinery. Comparative omics approaches can support rational genetic engineering and/or breeding of filamentous fungi with improved cellulase production capacity. In this study, comparative genomic, transcriptomic and secretomic profiling of P. oxalicum HP7-1 and its cellulase and xylanase hyper-producing mutant EU2106 were employed to screen for novel regulators of cellulase and xylanase gene expression.ResultsThe 30.62 Mb P. oxalicum HP7-1 genome was sequenced, and 9834 protein-coding genes were annotated. Re-sequencing of the mutant EU2106 genome identified 274 single nucleotide variations and 12 insertion/deletions. Comparative genomic, transcriptomic and secretomic profiling of HP7-1 and EU2106 revealed four candidate regulators of cellulase and xylanase gene expression. Deletion of these candidate genes and measurement of the enzymatic activity of the resultant mutants confirmed the identity of three regulatory genes. POX02484 and POX08522, encoding a putative Zn(II)2Cys6 DNA-binding domain and forkhead protein, respectively, were found to be novel, while PoxClrB is an ortholog of ClrB, a key transcriptional regulator of cellulolytic enzyme gene expression in filamentous fungi. ΔPOX02484 and ΔPOX08522 mutants exhibited significantly reduced β-glucosidase activity, increased carboxymethylcellulose cellulase and xylanase activities, and altered transcription level of cellulase and xylanase genes compared with the parent strain ΔPoxKu70, with Avicel as the sole carbon source.ConclusionsTwo novel genes, POX02484 and POX08522, were found and characterized to regulate the expression of cellulase and xylanase genes in P. oxalicum. These findings are important for engineering filamentous fungi to improve cellulase and xylanase production.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0616-9) contains supplementary material, which is available to authorized users.
BackgroundThe transition to a more environmentally friendly economy has prompted studies of modern biorefineries, including the utilization of low-value lignocellulose. The major challenge facing the widespread application of biorefineries is the high cost of enzymes that can efficiently hydrolyze recalcitrant cellulose to sugars. Penicillium oxalicum produces large amounts of plant-cell-wall-degrading enzymes, but their production is tightly controlled by complex regulatory networks, resulting in low yields of the native enzymes. Regulatory genes have been the targets of genetic engineering to improve enzyme production in microorganisms. In this study, we used transcriptomic profiling and genetic analyses to screen for and identify novel key regulators of cellulase and xylanase gene expression in P. oxalicum.ResultsA comparative analysis of the transcriptomes of P. oxalicum HP7-1 on different carbon sources, including glucose, wheat bran, and wheat bran plus Avicel, identified 40 candidate genes regulating the expression of cellulolytic enzyme genes. Deletion mutants of 31 candidate genes were constructed in P. oxalicum ∆PoxKu70 and 11 resultant mutants showed significant changes in their filter-paper cellulase production compared with the parental strain ∆PoxKu70. Among these 11 mutants, ΔPoxCxrA, ΔPoxCxrB, and ΔPoxNsdD showed the most significant reduction in the enzyme production (96.8, 75.9, and 58.5%, respectively). Ten of these 11 genes are here reported to be involved in cellulase production for the first time. Further tests revealed that ΔPoxCxrA, ΔPoxCxrB, and ΔPoxNsdD displayed significantly reduced xylanase production, whereas ΔPoxCxrA produced negligible xylanase. Interestingly, ΔPoxCxrB and ΔPoxNsdD showed significantly increased β-glucosidase production. Real-time quantitative reverse transcription–PCR and an electrophoretic mobility shift assay (EMSA) showed that PoxCxrA, PoxCxrB, and PoxNsdD regulate the expression of one another, but the mode of regulation changes dynamically during the growth of fungal cells in the presence of cellulose. EMSA showed that PoxCxrA, PoxCxrB, and PoxNsdD directly bind the putative promoters of major cellulase and xylanase genes.ConclusionsWe have detected and identified three key new regulatory genes, PoxCxrA, PoxCxrB, and PoxNsdD, that directly and indirectly regulate the expression of cellulase and xylanase genes in P. oxalicum. This study provides novel insights into the regulatory mechanisms of fungal cellulase and xylanase gene expression.Electronic supplementary materialThe online version of this article (10.1186/s13068-017-0966-y) contains supplementary material, which is available to authorized users.
Microwave absorbing materials with high absorption over a broad bandwidth when they have a small thickness are strongly desired due to their widespread applications. Herein, cerium oxide immobilized reduced graphene oxide (CeO2-rGO) hybrids with excellent microwave absorbing performance have been fabricated by a versatile one-step hydrothermal approach. Modern measurement techniques, including X-ray diffraction, Raman spectroscopy, electronic microscopy, X-ray photoelectron spectroscopy and vector network analysis, have been conducted to characterize the chemical composition, microstructure and electromagnetic performance of the as-obtained hybrids. Morphological analysis reveals that the CeO2 nanocrystals are homogeneously immobilized onto the rGO surface without any significant agglomeration. Interestingly, significant enhancement in the microwave absorbing performance has been observed for all the CeO2-rGO hybrids. For example, a CeO2-rGO hybrid with a 10 : 1 mass ratio of CeO2 to GO exhibits a minimum reflection loss (RL) of -45.94 dB, which is 73.35 times and 6.14 times that of the lone CeO2 and rGO, respectively. Moreover, the CeO2-rGO hybrid shows a broadband absorption feature with an effective absorption bandwidth (RL < -10 dB) of 4.5 GHz, and can be exploited for practical application in a frequency range of 3.68-18.00 GHz via tuning of the thickness. Investigation of the structure-property correlation indicates that such enhancements are attributed to conductive loss, polarization loss and multiple reflections which are mainly derived from the unique CeO2-rGO based architecture. In addition, the higher oxygen vacancy concentration of CeO2 in hybrids can promote electron transfer between CeO2 and rGO, leading to microwave attenuation enhancement. It is expected that these CeO2-rGO hybrids can be used as new microwave absorbers.
Soil fungi produce a wide range of chemical compounds and enzymes with potential for applications in medicine and biotechnology. Cellular processes in soil fungi are highly dependent on the regulation under environmentally induced stress, but most of the underlying mechanisms remain unclear. Previous work identified a key GATA-type transcription factor, NsdD (PoxNsdD; also called POX08415), that regulates the expression of cellulase and xylanase genes in PoxNsdD shares 57 to 64% identity with the key activator NsdD, involved in asexual development in In the present study, the regulatory roles of PoxNsdD in were further explored. Comparative transcriptomic profiling revealed that PoxNsdD regulates major genes involved in starch, cellulose, and hemicellulose degradation, as well as conidiation and pigment biosynthesis. Subsequent experiments confirmed that a Δ strain lost 43.9 to 78.8% of starch-digesting enzyme activity when grown on soluble corn starch, and it produced 54.9 to 146.0% more conidia than the Δ parental strain. During cultivation, Δ cultures changed color, from pale orange to brick red, while the Δ cultures remained bluish white. Real-time quantitative reverse transcription-PCR showed that dynamically regulated the expression of a glucoamylase gene (/), an α-amylase gene (/), and a regulatory gene (/), as well as a polyketide synthase gene (//) for yellow pigment biosynthesis and a conidiation-regulated gene (/). Moreover, binding experiments showed that PoxNsdD bound the promoter regions of the above-described genes. This work provides novel insights into the regulatory mechanisms of fungal cellular processes and may assist in genetic engineering of for potential industrial and medical applications. Most filamentous fungi produce a vast number of extracellular enzymes that are used commercially for biorefineries of plant biomass to produce biofuels and value-added chemicals, which might promote the transition to a more environmentally friendly economy. The expression of these extracellular enzyme genes is tightly controlled at the transcriptional level, which limits their yields. Hitherto our understanding of the regulation of expression of plant biomass-degrading enzyme genes in filamentous fungi has been rather limited. In the present study, regulatory roles of a key regulator, PoxNsdD, were further explored in the soil fungus , contributing to the understanding of gene regulation in filamentous fungi and revealing the biotechnological potential of via genetic engineering.
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