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1.
PLoS Genet ; 19(7): e1010867, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37523410

RESUMO

Many filamentous fungi produce plant-polysaccharide-degrading enzymes (PPDE); however, the regulatory mechanism of this process is poorly understood. A Gal4-like transcription factor, CxrA, is essential for mycelial growth and PPDE production in Penicillium oxalicum. Its N-terminal region, CxrAΔ207-733 is required for the regulatory functions of whole CxrA, and contains a DNA-binding domain (CxrAΔ1-16&Δ59-733) and a methylated arginine (R) 94. Methylation of R94 is mediated by an arginine N-methyltransferase, PRMT2 and appears to induce dimerization of CxrAΔ1-60. Overexpression of prmt2 in P. oxalicum increases PPDE production by 41.4-95.1% during growth on Avicel, compared with the background strain Δku70;hphR+. Another arginine N-methyltransferase, PRMT3, appears to assist entry of CxrA into the nucleus, and interacts with CxrAΔ1-60 in vitro under Avicel induction. Deletion of prmt3 resulted in 67.0-149.7% enhanced PPDE production by P. oxalicum. These findings provide novel insights into the regulatory mechanism of fungal PPDE production.


Assuntos
Penicillium , Proteína-Arginina N-Metiltransferases , Proteína-Arginina N-Metiltransferases/genética , Penicillium/genética , Celulose , Arginina
2.
J Proteome Res ; 23(1): 449-464, 2024 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-38109854

RESUMO

Cancer's high incidence and death rate jeopardize human health and life, and it has become a global public health issue. Some members of NPCs have been studied in a few cancers, but comprehensive and prognostic analysis is lacking in most cancers. In this study, we used the Cancer Genome Atlas (TCGA) data genomics and transcriptome technology to examine the differential expression and prognosis of NPCs in 33 cancer samples, as well as to investigate NPCs mutations and their effect on patient prognosis and to evaluate the methylation level of NPCs in cancer. The linked mechanisms and medication resistance were subsequently investigated in order to investigate prospective tumor therapy approaches. The relationships between NPCs and immune infiltration, immune cells, immunological regulatory substances, and immune pathways were also investigated. Finally, the LUAD and KICH prognostic prediction models were built using univariate and multivariate COX regression analysis. Additionally, the mRNA and protein levels of NPCs were also identified.


Assuntos
Neoplasias Pulmonares , Neoplasias , Humanos , Estudos Prospectivos , Genômica , Análise Multivariada , Mutação , Neoplasias/genética , Prognóstico , Proteína C1 de Niemann-Pick , Proteínas de Transporte Vesicular , Proteínas de Membrana Transportadoras
3.
Dig Dis Sci ; 69(3): 1035-1054, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38282187

RESUMO

BACKGROUND: Liver hepatocellular carcinoma (LIHC) is a serious liver disease worldwide, and its pathogenesis is complicated. AIMS: This study investigated the potential role of FANCA in the advancement and prognosis of LIHC. METHODS: Public databases, quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blot (WB) and immunohistochemistry (IHC) were employed to measure FANCA expression between tumor and normal samples. The relationship between FANCA expression and prognosis of LIHC patients were examined. Functional enrichment of FANCA-related genes was performed. Furthermore, univariate and multivariate analyses were conducted to determine the independent prognosis value of FANCA in LIHC. Finally, influence of FANCA knockout on the proliferation, migration, and invasion of HepG2 cell was validated with cloning formation, CCK8, and Transwell assays. RESULTS: Expression analysis presented that FANCA had high expression level in LIHC tissues and cells. Receiver operating characteristic (ROC) curve analysis showed that FANCA was of great diagnosis value in LIHC. Clinicopathological analysis revealed that FANCA was significantly greater expressed in the advanced stage than in the early stage of LIHC. Univariate, multivariate, and Kaplan-Meier survival analysis confirmed that high expression of FANCA was strongly associated with poor survival of LIHC patients. In addition, high level of FANCA in LIHC showed a negative association with immunoinfiltrated B cells, T cells, and stromal scores. Moreover, Knockout of FANCA significantly inhibited HepG2 cell proliferative activity, migration, and invasion ability. CONCLUSIONS: Our data revealed that high level of FANCA was closely associated with LIHC malignant progression, suggesting its potential utility as a diagnostic, predictive indicator, and therapeutic target.


Assuntos
Carcinoma Hepatocelular , Anemia de Fanconi , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Western Blotting , Prognóstico , Proteína do Grupo de Complementação A da Anemia de Fanconi/genética
4.
Funct Integr Genomics ; 23(3): 264, 2023 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-37541978

RESUMO

Liver cancer is a cunning malignancy with a high incidence and mortality rate among cancers worldwide. The NPC gene family members (NPCs: NPC1, NPC2, and NPC1L1) are closely linked to the development of multiple cancers, but their role in liver cancer remains unclear. As a result, we must investigate their functions in liver hepatocellular carcinoma (LIHC). NPCs were significantly differentially expressed between normal and LIHC tissues, with a high mutation frequency in LIHC. The ROC curve analysis revealed that NPC1/NPC2 had high diagnostic and prognostic values in LIHC. NPC1 expression was also found to be negatively correlated with its methylation level. The differentially expressed genes between high and low NPC1 expression groups in LIHC were mainly related to channel activity, transporter complexes, and plasma membrane adhesion molecules. Additionally, NPC1 expression was significantly associated with multiple immune cells and immunization checkpoints. It was hypothesized that a TUG1/SNHG4-miR-148a-3p-NPC1 regulatory axis is associated with hepatocarcinogenesis. Finally, the protein expression of NPC1 in LIHC tissues and paraneoplastic tissues was detected, and NPC1-knockdown HepG2 cells (NPC1KO) inhibited the proliferation, migration, and invasion. This study helped to identify new prognostic markers and potential immunotherapeutic targets for LIHC and revealed the molecular mechanisms underlying NPC1 regulation in LIHC. The NPCs play a key role in the prognosis and diagnosis of LIHC and may be an important indicator for LIHC prognosis and diagnosis; NPC1 might be a potential therapeutic target in LIHC.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/genética , Prognóstico , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/genética , Multiômica
5.
Biochem Genet ; 2023 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-37658254

RESUMO

Metabolites are important indicators of cancer and mutations in genes involved in amino acid metabolism may influence tumorigenesis. Immunotherapy is an effective cancer treatment option; however, its relationship with amino acid metabolism has not been reported. In this study, RNA-seq data for 371 liver cancer patients were acquired from TCGA and used as the training set. Data for 231 liver cancer patients were obtained from ICGC and used as the validation set to establish a gene signature for predicting liver cancer overall survival outcomes and immunotherapeutic responses. Four reliable groups based on 132 amino acid metabolism-related DEGs were obtained by consistent clustering of 371 HCC patients and a four-gene signature for prediction of liver cancer survival outcomes was developed. Our data show that in different clinical groups, the overall survival outcomes in the high-risk group were markedly low relative to the low-risk group. Univariate and multivariate analyses revealed that the characteristics of the 4-gene signature were independent prognostic factors for liver cancer. The ROC curve revealed that the risk characteristic is an efficient predictor for 1-, 2-, and 3-year HCC survival outcomes. The GSVA and KEGG pathway analyses revealed that high-risk score tumors were associated with all aspects of the degree of malignancy in liver cancer. There were more mutant genes and greater immune infiltrations in the high-risk groups. Assessment of the three immunotherapeutic cohorts established that low-risk score patients significantly benefited from immunotherapy. Then, we established a prognostic nomogram based on the TCGA cohort. In conclusion, the 4-gene signature is a reliable diagnostic marker and predictor for immunotherapeutic efficacy.

6.
Mol Microbiol ; 116(1): 140-153, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33561892

RESUMO

The essential transcription factor PoxCxrA is required for cellulase and xylanase gene expression in the filamentous fungus Penicillium oxalicum that is potentially applied in biotechnological industry as a result of the existence of the integrated cellulolytic and xylolytic system. However, the regulatory mechanism of cellulase and xylanase gene expression specifically associated with PoxCxrA regulation in fungi is poorly understood. In this study, the novel regulator PoxCbh (POX06865), containing a centromere protein B-type helix-turn-helix domain, was identified through screening for the PoxCxrA regulon under Avicel induction and genetic analysis. The mutant ∆PoxCbh showed significant reduction in cellulase and xylanase production, ranging from 28.4% to 59.8%. Furthermore, PoxCbh was found to directly regulate the expression of important cellulase and xylanase genes, as well as the known regulatory genes PoxNsdD and POX02484, and its expression was directly controlled by PoxCxrA. The PoxCbh-binding DNA sequence in the promoter region of the cellobiohydrolase 1 gene cbh1 was identified. These results expand our understanding of the diverse roles of centromere protein B-like protein, the regulatory network of cellulase and xylanase gene expression, and regulatory mechanisms in fungi.


Assuntos
Proteína B de Centrômero/genética , Proteínas Cromossômicas não Histona/biossíntese , Regulação Fúngica da Expressão Gênica/genética , Sequências Hélice-Volta-Hélice/genética , Penicillium/genética , Penicillium/metabolismo , Celulase/biossíntese , Celulase/genética , Celulose 1,4-beta-Celobiosidase/genética , Proteína B de Centrômero/biossíntese , Proteínas Cromossômicas não Histona/genética , Endo-1,4-beta-Xilanases/biossíntese , Endo-1,4-beta-Xilanases/genética , Fatores de Transcrição/genética
7.
Int J Mol Sci ; 23(16)2022 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-36012145

RESUMO

IAN873, Dongfang93114 and Reyan73397, created through vegetative propagation for their high yield and excellent cold resistance, are major clones planted in China. In this work, latexes with rubber particles of the same size from these clones are separated from fresh natural rubber latex, and corresponding rubber films are prepared from each latex. The structure and components of each film are measured. This indicates that the characteristics of the rubbers obtained from latexes with similar particle sizes show some resembling trends among different clones, while for specific samples, those characteristics vary depending on the clone. The molecular weight is generally highest in IAN873 and lowest in Reyan73397. Rubber chains in small rubber particles are longer, and large rubber particles show a wider molecular weight distribution. The gel content of every sample from Reyan73397 is lower than the other two clones. The nitrogen content increases with the size of rubber particles in all clones. The ester content of small rubber particles in IAN873 and Reyan73397 is almost zero. Large rubber particles have more branching points formed via esters. This study provides a new perspective on the influence of clones on the relationship between characteristics of natural rubber and the size of rubber particles in natural rubber latex.


Assuntos
Látex , Borracha , Células Clonais , Látex/química , Peso Molecular , Proteínas de Plantas/química , Plantas
8.
Appl Microbiol Biotechnol ; 105(11): 4675-4691, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34076714

RESUMO

Heterotrimeric-G-protein-mediated signaling pathways modulate the expression of the essential genes in many fundamental cellular processes in fungi at the transcription level. However, these processes remain unclear in Penicillium oxalicum. In this study, we generated knockout and knockout-complemented strains of gng-1 (POX07071) encoding the Gγ protein and found that GNG-1 modulated the expression of genes encoding plant-biomass-degrading enzymes (PBDEs) and sporulation-related activators. Interestingly, GNG-1 affected expression of the cxrB that encodes a known transcription factor required for the expression of major cellulase and xylanase genes. Constitutive overexpression of cxrB in ∆gng-1 circumvented the dependence of PBDE production on GNG-1. Further evidence indicated that CxrB indirectly regulated the transcription levels of key amylase genes by controlling the expression of the regulatory gene amyR. These data extended the diversity of Gγ protein functions and provided new insight into the signal transduction and regulation of PBDE gene expression in filamentous fungi. KEY POINTS: • GNG-1 modulates the expression of PBDE genes and sporulation-related genes. • GNG-1 controls expression of the key regulatory gene cxrB. • Overexpression of cxrB circumvents dependence of PBDE production on GNG-1.


Assuntos
Subunidades gama da Proteína de Ligação ao GTP , Penicillium , Biomassa , Regulação Fúngica da Expressão Gênica , Penicillium/genética
9.
Appl Microbiol Biotechnol ; 105(2): 679-694, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33394158

RESUMO

Phospholipases play vital roles in immune and inflammatory responses in mammals and plants; however, knowledge of phospholipase functions in fungi is limited. In this study, we investigated the effects of deleting predicted phospholipase genes on cellulase and xylanase production, and morphological phenotype, in Penicillium oxalicum. Individual deletion of nine of the ten predicted phospholipase genes resulted in alteration of cellulase and xylanase production, and the morphological phenotypes, to various degrees. The mutant ∆POX07277 lost 22.5 to 82.8% of cellulase (i.e., filter paper cellulase, carboxymethylcellulase, and p-nitrophenyl-ß-cellobiosidase) and xylanase production, whereas p-nitrophenyl-ß-glucopyranosidase production increased by 5.8-127.8 fold. POX07277 (P. oxalicum gene No. 07277) was predicted to encode phospholipase A2 and was found to negatively affect the sporulation of P. oxalicum. Comparative transcriptomic and quantitative reverse transcription-PCR analysis indicated that POX07277 dynamically affected the expression of cellulase and xylanase genes and the regulatory genes for fungal sporulation, under micro-crystalline cellulose induction. POX07277 was required for the expression of the known regulatory gene PoxCxrB (cellulolytic and xylanolytic regulator B in P. oxalicum), which is involved in cellulase and xylanase gene expression in P. oxalicum. Conversely, POX07277 expression was regulated by PoxCxrB. These findings will aid the understanding of phospholipase functions and provide novel insights into the mechanism of fungal cellulase and xylanase gene expression. KEY POINTS : • The roles of phospholipases were investigated in Penicillium oxalicum. • POX07277 (PLA2) is required for the expression of cellulase and xylanase genes. • PoxCxrB dynamically regulated POX07277 expression.


Assuntos
Celulase/biossíntese , Endo-1,4-beta-Xilanases/biossíntese , Penicillium , Fosfolipases/metabolismo , Regulação Fúngica da Expressão Gênica , Penicillium/enzimologia , Penicillium/genética
10.
Appl Environ Microbiol ; 85(24)2019 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-31604764

RESUMO

Transcriptional regulation of cellulolytic and xylolytic genes in ascomycete fungi is controlled by specific carbon sources in different external environments. Here, comparative transcriptomic analyses of Penicillium oxalicum grown on wheat bran (WB), WB plus rice straw (WR), or WB plus Avicel (WA) as the sole carbon source under solid-state fermentation (SSF) revealed that most of the differentially expressed genes (DEGs) were involved in metabolism, specifically, carbohydrate metabolism. Of the DEGs, the basic core carbohydrate-active enzyme-encoding genes which responded to the plant biomass resources were identified in P. oxalicum, and their transcriptional levels changed to various extents depending on the different carbon sources. Moreover, this study found that three deletion mutants of genes encoding putative transcription factors showed significant alterations in filter paper cellulase production compared with that of a parental P. oxalicum strain with a deletion of Ku70 (ΔPoxKu70 strain) when grown on WR under SSF. Importantly, the ΔPoxAtf1 mutant (with a deletion of P. oxalicumAtf1, also called POX03016) displayed 46.1 to 183.2% more cellulase and xylanase production than a ΔPoxKu70 mutant after 2 days of growth on WR. RNA sequencing and quantitative reverse transcription-PCR revealed that PoxAtf1 dynamically regulated the expression of major cellulase and xylanase genes under SSF. PoxAtf1 bound to the promoter regions of the key cellulase and xylanase genes in vitro This study provides novel insights into the regulatory mechanism of fungal cellulase and xylanase gene expression under SSF.IMPORTANCE The transition to a more environmentally friendly economy encourages studies involving the high-value-added utilization of lignocellulosic biomass. Solid-state fermentation (SSF), that simulates the natural habitat of soil microorganisms, is used for a variety of applications such as biomass biorefinery. Prior to the current study, our understanding of genome-wide gene expression and of the regulation of gene expression of lignocellulose-degrading enzymes in ascomycete fungi during SSF was limited. Here, we employed RNA sequencing and genetic analyses to investigate transcriptomes of Penicillium oxalicum strain EU2101 cultured on medium containing different carbon sources and to identify and characterize transcription factors for regulating the expression of cellulase and xylanase genes during SSF. The results generated will provide novel insights into genetic engineering of filamentous fungi to further increase enzyme production.


Assuntos
Fator 1 Ativador da Transcrição/metabolismo , Ascomicetos/enzimologia , Ascomicetos/genética , Celulase/genética , Fermentação , Regulação Fúngica da Expressão Gênica , Xilosidases/genética , Ascomicetos/crescimento & desenvolvimento , Biomassa , Celulase/metabolismo , Meios de Cultura/química , DNA Fúngico/genética , Deleção de Genes , Genes Fúngicos/genética , Lignina/metabolismo , Penicillium/enzimologia , Penicillium/genética , Penicillium/crescimento & desenvolvimento , Regiões Promotoras Genéticas , RNA Fúngico/genética , Microbiologia do Solo , Xilosidases/metabolismo
11.
Appl Environ Microbiol ; 84(18)2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-29980558

RESUMO

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, Penicillium oxalicum NsdD (PoxNsdD; also called POX08415), that regulates the expression of cellulase and xylanase genes in P. oxalicum PoxNsdD shares 57 to 64% identity with the key activator NsdD, involved in asexual development in Aspergillus In the present study, the regulatory roles of PoxNsdD in P. oxalicum 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 ΔPoxNsdD 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 ΔPoxKu70 parental strain. During cultivation, ΔPoxNsdD cultures changed color, from pale orange to brick red, while the ΔPoxKu70 cultures remained bluish white. Real-time quantitative reverse transcription-PCR showed that PoxNsdD dynamically regulated the expression of a glucoamylase gene (POX01356/Amy15A), an α-amylase gene (POX09352/Amy13A), and a regulatory gene (POX03890/amyR), as well as a polyketide synthase gene (POX01430/alb1/wA) for yellow pigment biosynthesis and a conidiation-regulated gene (POX06534/brlA). Moreover, in vitro 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 Poxalicum for potential industrial and medical applications.IMPORTANCE 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 Penicillium oxalicum, contributing to the understanding of gene regulation in filamentous fungi and revealing the biotechnological potential of Poxalicum via genetic engineering.


Assuntos
Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Penicillium/metabolismo , Pigmentos Biológicos/biossíntese , Esporos Fúngicos/crescimento & desenvolvimento , Fatores de Transcrição/metabolismo , Biodegradação Ambiental , Celulase/genética , Celulase/metabolismo , Celulose/metabolismo , Proteínas Fúngicas/genética , Perfilação da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Glucana 1,4-alfa-Glucosidase/genética , Glucana 1,4-alfa-Glucosidase/metabolismo , Penicillium/enzimologia , Penicillium/genética , Penicillium/crescimento & desenvolvimento , Policetídeo Sintases/genética , Policetídeo Sintases/metabolismo , Esporos Fúngicos/genética , Esporos Fúngicos/metabolismo , Fatores de Transcrição/genética , alfa-Amilases/genética , alfa-Amilases/metabolismo
12.
Phys Chem Chem Phys ; 20(20): 14155-14165, 2018 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-29749422

RESUMO

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.

13.
Appl Microbiol Biotechnol ; 102(8): 3739-3753, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29511847

RESUMO

High-mobility group (HMG)-box proteins are involved in chromatin organization in eukaryotes, especially in sex determination and regulation of mitochondrial DNA compaction. Although a novel HMG-box protein, PoxHmbB, had been initially identified to be required for filter paper cellulase activity by Penicillium oxalicum, the biological roles of HMG-box proteins in biomass-degrading enzyme production have not been systematically explored. The P. oxalicum mutant ∆PoxHmbB lost 34.7-86.5% of cellulase (endoglucanase, p-nitrophenyl-ß-cellobiosidase, and p-nitrophenyl-ß-glucopyranosidase) activities and 60.3% of xylanase activity following Avicel induction, whereas it exhibited about onefold increase in amylase activity following soluble corn starch induction. Furthermore, ∆PoxHmbB presented delayed conidiation and hyphae growth. Transcriptomic profiling and real-time quantitative reverse transcription-PCR revealed that PoxHmbB regulated the expression of major genes encoding plant biomass-degrading enzymes such as PoxCel7A-2, PoxCel5B, PoxBgl3A, PoxXyn11B, and PoxGA15A, as well as those involved in conidiation such as PoxBrlA. In vitro binding experiments further confirmed that PoxHmbB directly binds to the promoter regions of these major genes. These results further indicate the diversity of the biological functions of HMG-box proteins and provide a novel and promising engineering target for improving plant biomass-degrading enzyme production in filamentous fungi.


Assuntos
Celulase/biossíntese , Celulase/genética , Proteínas HMGB/metabolismo , Penicillium/enzimologia , Penicillium/genética , Biomassa , Celulase/metabolismo , Regulação Fúngica da Expressão Gênica/genética , Mutação
14.
World J Microbiol Biotechnol ; 33(9): 171, 2017 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-28849313

RESUMO

Talaromyces pinophilus is a promising filamentous fungus for industrial production of biomass-degrading enzymes used in biorefining, and its genome was recently sequenced and reported. However, functional analysis of genes in T. pinophilus is rather limited owing to lack of genetic tools. In this study, a putative TpKu70 encoding the Ku70 homolog involved in the classic non-homologous end-joining pathway was deleted in T. pinophilus 1-95. ΔTpKu70 displayed no apparent defect in vegetative growth and enzyme production, and presented similar sensitivity to benomyl, bleomycin, and UV, when compared with the wild-type T. pinophilus strain 1-95. Seven genes that encode putative transcription factors, including TpAmyR, were successfully knocked out in ΔTpKu70 at 61.5-100% of homologous recombination frequency, which is significantly higher than that noted in the wild-type. Interestingly, ΔTpAmyR produced approximately 20% of amylase secreted by the parent strain ΔTpKu70 in medium containing soluble starch from corn as the sole carbon source. Real-time quantitative reverse transcription PCR showed that TpAmyR positively regulated the expression of genes encoding α-amylase and glucoamylase. Thus, this study provides a useful tool for genetic analysis of T. pinophilus, and identification of a key role for the transcription factor TpAmyR in amylase production in T. pinophilus.


Assuntos
Autoantígeno Ku/genética , Talaromyces/crescimento & desenvolvimento , Fatores de Transcrição/genética , Amilases/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Deleção de Genes , Técnicas de Inativação de Genes , Recombinação Homóloga , Talaromyces/enzimologia , Talaromyces/genética , Fatores de Transcrição/metabolismo
15.
Int J Biol Macromol ; 242(Pt 1): 124643, 2023 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-37119904

RESUMO

The knowledge of agarose (AG) oxidation using periodate as oxidizer has not been systematically explored. This paper synthesized oxidized agarose (OAG) using solid-sate and solution reaction methods; the reaction mechanism and the properties of OAG samples were systematically evaluated. Chemical structure analysis disclosed that the aldehyde group and carboxyl group contents in all OAG samples are extremely low. Meanwhile, crystallinity, dynamic viscosity and molecular weight of OAG samples is lower than that of the original AG. Reaction temperature, time and sodium periodate dosage are inversely proportional to the decline of the gelling temperature (Tg) and melting temperature (Tm); and the Tg and Tm for the OAG sample obtained are even 19 °C and 22 °C lower than that of the original AG. The as-synthesized OAG samples all possess excellent cytocompatibility and blood compatibility; and can promote the proliferation and migration of fibroblast cells. Last but not least, the gel strength, hardness, cohesiveness, springiness and chewiness of the OAG gel can be effectively regulated via oxidation reaction. In conclusions, both solid and solution oxidation can regulate the physical properties of OAG and enlarge its potential applications in wound dressing, tissue engineering and food areas.


Assuntos
Sefarose , Sefarose/química , Temperatura , Oxirredução , Viscosidade , Géis/química
16.
Polymers (Basel) ; 15(4)2023 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-36850139

RESUMO

Natural rubber (NR) exhibits good elasticity, flexural resistance, wear resistance, and excellent mechanical properties, and it has been widely used in aerospace, transportation, medical, and health fields. For NR, however, the resistance to thermal-oxidation and ozone aging is fairly poor. Although aging properties of NR can be significantly improved with the incorporation of chloroprene rubber (CR) according to some references, the miscibility between NR and CR, the morphologies of the binary blends, and so on are revealed ambiguously. In this work, molecular dynamics simulation (MD) and dissipative particle dynamics (DPD) simulation were carried out to predict the compatibility between natural rubber and chloroprene rubber in view of Flory-Huggins parameters. The morphologies of the blends were obtained with the use of the DPD method. The simulation results were furtherly examined by means of Fourier transform infrared spectroscopy (FT-IR) and dynamic mechanical analysis (DMA). It was found that the miscibility between NR and CR is poor. Nevertheless, the miscibility could be improved when the content of CR is 50% or 90%. In addition, spinodal decomposition with a critical temperature of 390 K would take place according to the phase diagram. Microphase structure such as spherical, lamellar, and bicontinuous phases can be found with different contents of CR in the blends with the results of morphologies analysis.

17.
Nanomaterials (Basel) ; 13(10)2023 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-37242060

RESUMO

This study details microwave-absorbing materials made of natural rubber/nitrile butadiene rubber (NR/NBR) blends with multi-walled carbon nanotubes (MWCNTs) and molybdenum disulfide (MoS2). The mechanical blending method and the influences of fabrication on the morphology and microwave-absorbing performance of resulting compounds were logically investigated. It was found that interfacial differences between the fillers and matrix promote the formation of MWCNTs and MoS2 networks in NR/NBR blends, thus improving microwave-absorbing performance. Compared with direct compounding, masterbatch-based two-step blending is more conducive to forming interpenetrating networks of MWCNTs/MoS2, endowing the resulting composite with better microwave attenuation capacity. Composites with MWCNTs in NR and MoS2 in NBR demonstrate the best microwave-absorbing performance, with a minimum reflection loss of -44.54 dB and an effective absorption bandwidth of 3.60 GHz. Exploring the relationship between morphology and electromagnetic loss behavior denotes that such improvement results from the selective distribution of dual fillers, inducing networking and multi-component-derived interfacial polarization enhancement.

18.
Polymers (Basel) ; 14(7)2022 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-35406253

RESUMO

In this paper, biodegradable epoxidized natural rubber containing cyclic carbonate groups (CNR) was prepared by the reaction between epoxidized natural rubber (ENR) and carbon dioxide. Dynamic disulfide bonds and a boronic ester structure were successfully constructed and then the cross-linking network was formed by the thermally initiated "click" reaction between thiol groups of the cross-linker and the residual epoxy groups of ENR. As a result of the exquisite double dynamic covalent structure, the material exhibits high self-healing efficiency. Moreover, by virtue of the cyclic carbonate structure of the CNR, the natural rubber was confirmed to be biodegradable according to the biodegradable measurement. To the best of our knowledge, natural rubber with biodegradable and self-healing characteristics was obtained for the first time.

19.
Materials (Basel) ; 15(11)2022 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-35683281

RESUMO

We propose a simple but rapid strategy to fabricate self-crosslinked dual-crosslinking elastomers (SCDCEs) with high mechanical properties. The SCDCEs are synthesized through one-pot copolymerization of Butyl acrylate (BA), acrylic amide (AM), and 3-Methacryloxypropyltrimethoxysilane (MEMO). Both the amino group on AM and the methoxy group on MEMO can be self-crosslinked after polymerization to form a dual-network crosslink consisting of hydrogen bonds crosslink and Si-O-Si covalent bonds crosslink. The SCDC endow optimal elastomer with high mechanical properties (the tensile strength is 6MPa and elongation at break is 490%) as the hydrogen bonds crosslink can serve as sacrificial construction to dissipate stress energy, while covalent crosslinking networks can ensure the elasticity and strength of the material. These two networks also contribute to the recoverability of the elastomers, leading them to recover their original shape and mechanical properties after being subjected to deformation in a short time.

20.
Materials (Basel) ; 15(14)2022 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-35888312

RESUMO

The design of hierarchical structures from biomass has become one of the hottest subjects in the field of microwave absorption due to its low cost, vast availability and sustainability. A kapok-fiber-derived carbon microtube was prepared by facile carbonization, and the relation between the structure and properties of the carbonized kapok fiber (CKF) was systematically investigated. The hollow tubular structures afford the resulting CKF composites with excellent microwave-absorbing performance. The sample with a 30 wt.% loading of CKF in paraffin demonstrates the strongest microwave attenuation capacity, with a minimum reflection loss of -49.46 dB at 16.48 GHz and 2.3 mm, and an optimized effective absorption bandwidth of 7.12 GHz (10.64-17.76 GHz, 2.3 mm) that covers 34% of the X-band and 96% of the Ku-band. Further, more than 90% of the incident electromagnetic wave in the frequency from 4.48 GHz to 18.00 GHz can be attenuated via tuning the thickness of the CKF-based absorber. This study outlines a foundation for the development of lightweight and sustainable microwave absorbers with a high absorption capacity and broad effective absorption bandwidth.

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