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Introduction: Streptococcus thermophilus is a traditional starter for dairy products. The lactose rich in milk is the main carbon source for the growth of S. thermophilus. However, the utilization of galactose by S. thermophilus is strain-specific, and many genetic factors can affect the sugar utilization phenotype of S. thermophilus strains. Methods: In this study, S. thermophilus A25, which is capable of utilizing lactose and galactose, was used as the starting strain to construct lactose permease-deficient mutant S. thermophilus ΔlacS. Subsequently, the complement vectors expressing complete lactose permease of S. thermophilus and its N-terminal 1–486 amino acid residues were constructed and transformed into S. thermophilus ΔlacS, respectively. Meanwhile, complement vectors expressing lactose permease and galactose/proton symporter of Escherichia coli were also constructed. Results and Discussion: Results showed that S. thermophilus ΔlacS lost the ability to utilize lactose and galactose. By measuring the growth of the recombinant strains, it was found that the strain expressing complete lactose permease of S. thermophilus recovered the growth ability in lactose and galactose medium, while the strain expressing N-terminal of lactose permease recovered the growth ability only in lactose medium. Furthermore, the transformation of S. thermophilus ΔlacS was not successful with the complement vector expressing E. coli lactose permease, while the strain expressing E. coli galactose/proton symporter could recover its growth ability in the galactose medium. These results suggest that the properties of sugar transporters play an important role in galactose utilization by S. thermophilus. [ABSTRACT FROM AUTHOR]

Glycoside hydrolases (GHs) are carbohydrate-active enzymes essential for many environmental (e.g., carbon cycling) and biotechnological (e.g., biofuels) processes. The complete processing of carbohydrates by bacteria requires many enzymes acting synergistically. Here, I investigated the clustered or scattered distribution of 406,337 GH-genes and their association with transporter genes identified in 15,640 completely sequenced bacterial genomes. Different bacterial lineages displayed conserved levels of clustered or scattered GH-genes, but overall, the GH-genes clustering was generally higher than in randomized genomes. In lineages with highly clustered GH-genes (e.g., Bacteroides, Paenibacillus), clustered genes shared the same orientation. These codirectional gene clusters potentially facilitate the genes' co-expression by allowing transcriptional read-through and, at least in some cases, forming operons. In several taxa, the GH-genes clustered with distinct types of transporter genes. The type of transporter genes and the distribution of the so-called GH:TR-genes clusters were conserved in selected lineages. Globally, the phylogenetically conserved clustering of the GH-genes with transporter genes highlights the central function of carbohydrate processing across bacterial lineages. In addition, in bacteria with the most identified GH-genes, the genomic adaptations for carbohydrate processing also mirrored the broad environmental origin of the sequenced strains (e.g., soil and mammal gut) suggesting that a combination of evolutionary history and the environment selects for the specific supragenic organization of the GH-genes supporting the carbohydrate processing in bacterial genomes. [ABSTRACT FROM AUTHOR]

Ustilaginoidea virens infects rice, causing rice false smut disease and reduced yields. During its growth, U. virens can also produce some toxins but less is known about the response mechanisms of the plant to U. virens toxins. U. virens toxins can inhibit the accumulation of total sugar in rice panicles. We used RNA sequencing to analyze the differential expression profile induced by infiltrating crude toxins into early growth-stage rice panicles. We compared the transcriptomes of the control and crude toxin-treated rice panicles and determined variable transcriptional responses under the action of the crude toxins. A total of 6,127 differentially expressed genes (DEGs) were identified. Among these genes, 3,150 were upregulated and 2,977 were downregulated. Gene Ontology (GO) and metabolic pathway enrichment analyses indicated that U. virens toxins mainly influenced glycometabolism, amino acid metabolism, and secondary metabolism of rice panicles. DEG analysis showed that the gene expression levels of 10 transcription factor families were significantly changed. Genes involved in phenylpropanoid biosynthesis, flavonoid biosynthesis, sugar transporters, and starch synthesis-related were significantly downregulated, including cytochrome P450, beta-glucosidase, CHS1, sucrose transporters, SWEETs, starch-branching enzymes, and UDP-glucose pyrophosphorylase. However, genes involved in programmed cell death (PCD) were significantly upregulated and contained cytochrome c, metacaspase, and protein kinase genes. The results indicate that U. virens toxins may act as the pathogenic factors to reduce stress resistance, disrupt total sugar accumulation and starch formation, and induce PCD. [ABSTRACT FROM AUTHOR]

Ustilaginoidea virens infects rice, causing rice false smut disease and reduced yields. During its growth, U. virens can also produce some toxins but less is known about the response mechanisms of the plant to U. virens toxins. U. virens toxins can inhibit the accumulation of total sugar in rice panicles. We used RNA sequencing to analyze the differential expression profile induced by infiltrating crude toxins into early growth-stage rice panicles. We compared the transcriptomes of the control and crude toxin-treated rice panicles and determined variable transcriptional responses under the action of the crude toxins. A total of 6,127 differentially expressed genes (DEGs) were identified. Among these genes, 3,150 were upregulated and 2,977 were downregulated. Gene Ontology (GO) and metabolic pathway enrichment analyses indicated that U. virens toxins mainly influenced glycometabolism, amino acid metabolism, and secondary metabolism of rice panicles. DEG analysis showed that the gene expression levels of 10 transcription factor families were significantly changed. Genes involved in phenylpropanoid biosynthesis, flavonoid biosynthesis, sugar transporters, and starch synthesis-related were significantly downregulated, including cytochrome P450, beta-glucosidase, CHS1, sucrose transporters, SWEETs, starch-branching enzymes, and UDP-glucose pyrophosphorylase. However, genes involved in programmed cell death (PCD) were significantly upregulated and contained cytochrome c, metacaspase, and protein kinase genes. The results indicate that U. virens toxins may act as the pathogenic factors to reduce stress resistance, disrupt total sugar accumulation and starch formation, and induce PCD.

The filamentous fungus Trichoderma reesei secretes large quantities of cellulases and hemicellulases that have found wide applications in industry. Compared with extensive studies on the mechanism controlling cellulase gene expression, less is known about the regulatory mechanism behind xylanase gene expression. Herein, several putative sugar transporter encoding genes that showed significant upregulation on xylan were identified in T. reesei. Deletion of one such gene, gat1, resulted in increased xylanase production but hardly affected cellulase induction. Further analyses demonstrated that deletion of gat1 markedly increased XYNI production at the transcriptional level and only exerted a minor effect on XYNII synthesis. In contrast, overexpressing gat1 caused a continuous decrease in xyn1 expression. Deletion of gat1 also affected the expression of xyn1 and pectinase genes when T. reesei was cultivated with galacturonic acid as the sole carbon source. Transcriptome analyses of Δgat1 and its parental strain identified 255 differentially expressed genes that are enriched in categories of glycoside hydrolases, lipid metabolism, transporters, and transcriptional factors. The results thus implicate a repressive role of the sugar transporter GAT1 in xyn1 expression and reveal that distinct regulatory mechanisms may exist in controlling the expression of different xylanase genes in T. reesei.

The filamentous fungus Trichoderma reesei secretes large quantities of cellulases and hemicellulases that have found wide applications in industry. Compared with extensive studies on the mechanism controlling cellulase gene expression, less is known about the regulatory mechanism behind xylanase gene expression. Herein, several putative sugar transporter encoding genes that showed significant upregulation on xylan were identified in T. reesei. Deletion of one such gene, gat1 , resulted in increased xylanase production but hardly affected cellulase induction. Further analyses demonstrated that deletion of gat1 markedly increased XYNI production at the transcriptional level and only exerted a minor effect on XYNII synthesis. In contrast, overexpressing gat1 caused a continuous decrease in xyn1 expression. Deletion of gat1 also affected the expression of xyn1 and pectinase genes when T. reesei was cultivated with galacturonic acid as the sole carbon source. Transcriptome analyses of Δ gat1 and its parental strain identified 255 differentially expressed genes that are enriched in categories of glycoside hydrolases, lipid metabolism, transporters, and transcriptional factors. The results thus implicate a repressive role of the sugar transporter GAT1 in xyn1 expression and reveal that distinct regulatory mechanisms may exist in controlling the expression of different xylanase genes in T. reesei. [ABSTRACT FROM AUTHOR]

Aspergillus niger is one of the most widely used fungi to study the conversion of the lignocellulosic feedstocks into fermentable sugars. Understanding the sugar uptake system of A. niger is essential to improve the efficiency of the process of fungal plant biomass degradation. In this study, we report a comprehensive characterization of the sugar transportome of A. niger by combining phylogenetic and comparative transcriptomic analyses. We identified 86 putative sugar transporter (ST) genes based on a conserved protein domain search. All these candidates were then classified into nine subfamilies and their functional motifs and possible sugar-specificity were annotated according to phylogenetic analysis and literature mining. Furthermore, we comparatively analyzed the ST gene expression on a large set of fungal growth conditions including mono-, di- and polysaccharides, and mutants of transcriptional regulators. This revealed that transporter genes from the same phylogenetic clade displayed very diverse expression patterns and were regulated by different transcriptional factors. The genome-wide study of STs of A. niger provides new insights into the mechanisms underlying an extremely flexible metabolism and high nutritional versatility of A. niger and will facilitate further biochemical characterization and industrial applications of these candidate STs.

Xylan constituted with β-1,4-D-xylose linked backbone and diverse substituted side-chains is the most abundant hemicellulose component of biomass, which can be completely and rapidly degraded into fermentable sugars by Aspergillus niger. This is of great value for obtaining renewable biofuels and biochemicals. To clarify the underlying mechanisms associated with highly efficient xylan degradation, assimilation, and metabolism by A. niger, we utilized functional proteomics to analyze the secreted proteins, sugar transporters, and intracellular proteins of A. niger An76 grown on xylan-based substrates. Results demonstrated that the complete xylanolytic enzyme system required for xylan degradation and composed of diverse isozymes was secreted in a sequential order. Xylan-backbone-degrading enzymes were preferentially induced by xylose or other soluble sugars, which efficiently produced large amounts of xylooligosaccharides (XOS) and xylose; however, XOS was more efficient than xylose in triggering the expression of the key transcription activator XlnR, resulting in higher xylanase activity and shortening xylanase-production time. Moreover, the substituted XOS was responsible for improving the abundance of side-chain-degrading enzymes, specific transporters, and key reductases and dehydrogenases in the pentose catabolic pathway. Our findings indicated that industries might be able to improve the species and concentrations of xylan-degrading enzymes and shorten fermentation time by adding abundant intermediate products of natural xylan (XOS) to cultures of filamentous fungi.

Xylan constituted with b-1,4-D-xylose linked backbone and diverse substituted side-chains is the most abundant hemicellulose component of biomass, which can be completely and rapidly degraded into fermentable sugars by Aspergillus niger. This is of great value for obtaining renewable biofuels and biochemicals. To clarify the underlying mechanisms associated with highly efficient xylan degradation, assimilation, and metabolism by A. niger, we utilized functional proteomics to analyze the secreted proteins, sugar transporters, and intracellular proteins of A. niger An76 grown on xylan-based substrates. Results demonstrated that the complete xylanolytic enzyme system required for xylan degradation and composed of diverse isozymes was secreted in a sequential order. Xylan-backbone-degrading enzymes were preferentially induced by xylose or other soluble sugars, which efficiently produced large amounts of xylooligosaccharides (XOS) and xylose; however, XOS was more efficient than xylose in triggering the expression of the key transcription activator XlnR, resulting in higher xylanase activity and shortening xylanase-production time. Moreover, the substituted XOS was responsible for improving the abundance of side-chain-degrading enzymes, specific transporters, and key reductases and dehydrogenases in the pentose catabolic pathway. Our findings indicated that industries might be able to improve the species and concentrations of xylan-degrading enzymes and shorten fermentation time by adding abundant intermediate products of natural xylan (XOS) to cultures of filamentous fungi. [ABSTRACT FROM AUTHOR]