The improvement of bacterial tolerance to organic solvents is a main prerequisite for the microbial production of biofuels which are toxic to cells. For targeted genetic engineering of Escherichia coli to increase organic solvent tolerances (OSTs), we selected and investigated a total of 12 genes that participate in relevant mechanisms to tolerance. In a spot assay of 12 knockout mutants with n-hexane and cyclohexane, the genes fadR and marR were finally selected as the two key genes for engineering. Fatty acid degradation regulon (FadR) regulates the biosynthesis and degradation of fatty acids coordinately, and the multiple antibiotic resistance repressor (MarR) is the repressor of the global regulator MarA for multidrug resistance. In the competitive growth assay, the ΔmarR mutant became dominant when the pooled culture of 11 knockout mutants was cultivated successively in the presence of organic solvent. The increased OSTs in the ΔmarR and ΔfadR mutants were confirmed by a growth experiment and a viability test. The even more highly enhanced OSTs in the ΔfadR ΔmarR double mutant were shown compared with the two single mutants. Cellular fatty acid analysis showed that the high ratio of saturated fatty acids to unsaturated fatty acids plays a crucial role in OSTs. Furthermore, the intracellular accumulation of OST strains was significantly decreased compared with the wild-type strain.
Background: Laminin-332 derived from keratinocytes plays a critical role in adhesion-related cell functions in melanocytes. Results: Keratinocyte-derived laminin-332 promotes the uptake of extracellular tyrosine and subsequent melanin synthesis in melanoma cells and melanocytes. Conclusion: Keratinocyte-derived laminin-332 promotes melanogenesis by controlling the uptake of tyrosine into melanocytes. Significance: Our finding reports novel means for regulating melanogenesis by the insoluble extracellular protein laminin-332.
Actinobacillus succinogenes, which is known to produce large amounts of succinate during fermentation of hexoses, was able to grow on C4-dicarboxylates such as fumarate under aerobic and anaerobic conditions. Anaerobic growth on fumarate was stimulated by glycerol and the major product was succinate, indicating the involvement of fumarate respiration similar to succinate production from glucose. The aerobic growth on C4-dicarboxylates and the transport proteins involved were studied. Fumarate was oxidized to acetate. The genome of A. succinogenes encodes six proteins with similarity to secondary C4-dicarboxylate transporters, including transporters of the Dcu (C4-dicarboxylate uptake), DcuC (C4-dicarboxylate uptake C), DASS (divalent anion : sodium symporter) and TDT (tellurite resistance dicarboxylate transporter) family. From the cloned genes, Asuc_0304 of the DASS family protein was able to restore aerobic growth on C4-dicarboxylates in a C4-dicarboxylate-transport-negative Escherichia coli strain. The strain regained succinate or fumarate uptake, which was dependent on the electrochemical proton potential and the presence of Na+. The transport had an optimum pH ~7, indicating transport of the dianionic C4-dicarboxylates. Transport competition experiments suggested substrate specificity for fumarate and succinate. The transport characteristics for C4-dicarboxylate uptake by cells of aerobically grown A. succinogenes were similar to those of Asuc_0304 expressed in E. coli, suggesting that Asuc_0304 has an important role in aerobic fumarate uptake in A. succinogenes. Asuc_0304 has sequence similarity to bacterial Na+-dicarboxylate cotransporters and contains the carboxylate-binding signature. Asuc_0304 was named SdcA (sodium-coupled C4-dicarboxylate transporter from A . succinogenes).
Volatile organic compounds (VOCs) are harmful to human health and the environment. Recently, loess (Hwangtoh) was used as an eco-friendly interior paint formulation in Korea. It is used even more commonly as a filter carrier to remove VOCs. In this study, we isolated Bacillus strains from a loess filter. The strains that were tolerant to VOCs were labeled according to the series VOC01 to VOC35. Four strains-VOC03, VOC11, VOC18, and VOC30-were investigated for their ability to degrade cyclohexane and toluene. Strain VOC18 best degraded both VOCs, whereas VOC03 demonstrated no ability to degrade VOCs. In keeping with this, VOC18 grew best on cyclohexane or toluene as the sole carbon source. The strains were identified by their physiochemical and phylogenetic characteristics. Strain VOC18 was determined as a strain of Bacillus cereus; VOC11 and VOC30 were determined as differentiated strains of B. thuringiensis. Strain VOC03, which demonstrated high tolerance but no ability to degrade VOCs, was identified as a strain of B. megaterium.
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