Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible...Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible for transcriptional regulation and the model of their interplay in induction and repression will be presented. Using in vivo foot printing analysis of xylan-induced and glucose repressed mycelia, we detected three adjacent nucleotide sequences contacted by DNA-binding proteins. Protection within the inverted repeat of the Cre1 (SYGGRG) consensus sequence on the non coding strand under repressing conditions is in perfect agreement with the previously reported Cre1 dependent glucose repression of xyn1. Constitutive protein binding could be observed to a CCAAT-box and an inverted repeat of a 5′ GGCTAA 3′ sequence. EMSA with crude extracts from induced and repressed mycelia revealed that the latter motifs are sufficient for formation of the basal transcriptional complex under all conditions. The inverted repeat of GGCTAA closely resembles the consensus sequences of the cellulase and xylanase regulators Ace1, Ace2 and, Xyr1 (encoded by xyr1, cloned and characterised in this study) EMSA with heterologously expressed components of each factor and of the T. reesei Hap2/3/5 protein complex revealed that the basal transcriptional complex is formed by Xyr1 and the Hap2/3/5. Additionally to the Cre1 mediated carbon catabolite repression a yet unknown mechanism antagonizing induction of xyn1 expression could be elucidated. Latter occurs through competition of the repressor Ace1 and Xyr1 for the GGCTAA motif. In vivo proof for the relevance of identified motifs could be given through analysis of T. reesei transformants containing correspondingly mutated versions of the xyn1 promoter fused to the A. niger goxA gene. The results indicated that the basal as well as the induction level of xyn1 gene transcription is dependent on an interaction of Xyr1 with the GGCTAA motif while formation of the CCAAT-Hap2/3/5 complex slightly reduces induction. It can be concluded that mutations impairing protein binding in vitro lead to a loss of distinct regulatory functions in xyn1 gene expression in vivo. A respective model of gene regulation will be presented.展开更多
A newly isolated bacterium was screened out for its survival on medium with 6 000 mg/L kraft lignin as the sole carbon source and energy, and identified as Pandoraea sp. by 16S rRNA gene sequence analysis. The biodegr...A newly isolated bacterium was screened out for its survival on medium with 6 000 mg/L kraft lignin as the sole carbon source and energy, and identified as Pandoraea sp. by 16S rRNA gene sequence analysis. The biodegradation experiment was carried out in mineral salt medium, containing 5 000 mg/L kraft lignin as only carbon and energy at pH 8.0 and 30℃. Under these conditions, significant reduction in color and lignin content by the strain was observed after incubation for 5 d. The strain attains maximum reduction capability in color （44.6%） and lignin content （39.9%） within 5 d of incubation, and reduced chemical oxygen demand（COD） from initial concentration 7 399 to 3 980 mg/L at maximum reduction level of 46.2% on the 4th day. The total ion chromatograph （TIC） of compounds presented in the chloroform extract of control and bacterial treated samples shows the formation of several lignin-related aromatic compounds including some small molecular lignin fragments, indicating a strong destruction in the lignin structure.展开更多
To shorten the bioleaching cycle of arsenic-containing gold concentrate, surfactants were used to promote the interaction between bacteria and ore to increase the arsenic leaching rate. Three different kinds of surfac...To shorten the bioleaching cycle of arsenic-containing gold concentrate, surfactants were used to promote the interaction between bacteria and ore to increase the arsenic leaching rate. Three different kinds of surfactants were used to evaluate the effects of surfactants on the growth of bacteria and arsenic leaching rate of arsenic-containing gold concentrate. The mechanism underlying surfactant enhancement was also studied. Results show that when relatively low-concentration surfactants are added to the medium, no significant difference is observed in the growth and Fe2+ oxidation ability of the bacteria compared with no surfactant in the medium. However, only the anionic surfactant calcium lignosulfonate and the nonionic surfactant Tween 80 are found to improve the arsenic leaching rates. Their optimum mass concentrations are 30 and 80 mg/L, respectively. At such optimum mass concentrations, the arsenic leaching rates are approximately 13.7% and 9.1% higher than those without the addition of surfactant, respectively. Mechanism research reveals that adding the anionic surfactant calcium lignosulfonate improves the percentage of bacterial adhesion on the mineral surface and decreases the surface tension in the leaching solution.展开更多
文摘Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible for transcriptional regulation and the model of their interplay in induction and repression will be presented. Using in vivo foot printing analysis of xylan-induced and glucose repressed mycelia, we detected three adjacent nucleotide sequences contacted by DNA-binding proteins. Protection within the inverted repeat of the Cre1 (SYGGRG) consensus sequence on the non coding strand under repressing conditions is in perfect agreement with the previously reported Cre1 dependent glucose repression of xyn1. Constitutive protein binding could be observed to a CCAAT-box and an inverted repeat of a 5′ GGCTAA 3′ sequence. EMSA with crude extracts from induced and repressed mycelia revealed that the latter motifs are sufficient for formation of the basal transcriptional complex under all conditions. The inverted repeat of GGCTAA closely resembles the consensus sequences of the cellulase and xylanase regulators Ace1, Ace2 and, Xyr1 (encoded by xyr1, cloned and characterised in this study) EMSA with heterologously expressed components of each factor and of the T. reesei Hap2/3/5 protein complex revealed that the basal transcriptional complex is formed by Xyr1 and the Hap2/3/5. Additionally to the Cre1 mediated carbon catabolite repression a yet unknown mechanism antagonizing induction of xyn1 expression could be elucidated. Latter occurs through competition of the repressor Ace1 and Xyr1 for the GGCTAA motif. In vivo proof for the relevance of identified motifs could be given through analysis of T. reesei transformants containing correspondingly mutated versions of the xyn1 promoter fused to the A. niger goxA gene. The results indicated that the basal as well as the induction level of xyn1 gene transcription is dependent on an interaction of Xyr1 with the GGCTAA motif while formation of the CCAAT-Hap2/3/5 complex slightly reduces induction. It can be concluded that mutations impairing protein binding in vitro lead to a loss of distinct regulatory functions in xyn1 gene expression in vivo. A respective model of gene regulation will be presented.
基金Project(50925417) supported by the National Funds for Distinguished Young Scientists,ChinaProject(50830301) supported by the Key program of the National Natural Science Foundation of ChinaProject(2012BAC12B03) supported by the National 12th Five-year Plan for Science and Technology Support,China
文摘A newly isolated bacterium was screened out for its survival on medium with 6 000 mg/L kraft lignin as the sole carbon source and energy, and identified as Pandoraea sp. by 16S rRNA gene sequence analysis. The biodegradation experiment was carried out in mineral salt medium, containing 5 000 mg/L kraft lignin as only carbon and energy at pH 8.0 and 30℃. Under these conditions, significant reduction in color and lignin content by the strain was observed after incubation for 5 d. The strain attains maximum reduction capability in color （44.6%） and lignin content （39.9%） within 5 d of incubation, and reduced chemical oxygen demand（COD） from initial concentration 7 399 to 3 980 mg/L at maximum reduction level of 46.2% on the 4th day. The total ion chromatograph （TIC） of compounds presented in the chloroform extract of control and bacterial treated samples shows the formation of several lignin-related aromatic compounds including some small molecular lignin fragments, indicating a strong destruction in the lignin structure.
基金Projects(51104024,51374043)supported by National Natural Science Foundation of ChinaProject(10JJ6019)supported by Hunan Provincial Natural Science Foundation,China+1 种基金Project(10C0399)supported by Scientific Research Fund of Hunan Provincial Education Department,ChinaProject(2014SK3182)supported by Hunan Provincial Science&Technology Department,China
文摘To shorten the bioleaching cycle of arsenic-containing gold concentrate, surfactants were used to promote the interaction between bacteria and ore to increase the arsenic leaching rate. Three different kinds of surfactants were used to evaluate the effects of surfactants on the growth of bacteria and arsenic leaching rate of arsenic-containing gold concentrate. The mechanism underlying surfactant enhancement was also studied. Results show that when relatively low-concentration surfactants are added to the medium, no significant difference is observed in the growth and Fe2+ oxidation ability of the bacteria compared with no surfactant in the medium. However, only the anionic surfactant calcium lignosulfonate and the nonionic surfactant Tween 80 are found to improve the arsenic leaching rates. Their optimum mass concentrations are 30 and 80 mg/L, respectively. At such optimum mass concentrations, the arsenic leaching rates are approximately 13.7% and 9.1% higher than those without the addition of surfactant, respectively. Mechanism research reveals that adding the anionic surfactant calcium lignosulfonate improves the percentage of bacterial adhesion on the mineral surface and decreases the surface tension in the leaching solution.
