Improvement in the amino-acid microbial production

Amino-acid microbial production can be improved by a process modification or by inducing changes in the metabolism of the biocatalyst. These two approaches will be illustrated and discussed using the examples of glutamic acid and valine productions with Corynebacterium glutamicum.La production d'acide aminé par voie microbienne peut être améliorée soit en jouant sur le procédé de culture soit en modifiant le métabolisme du micro-organisme producteur. Ces 2 approches seront illustrées et discutées grâce à deux exemples d'acides aminés produits par Coryne-bacterium glutamicum: l'acide glutamique et la valine

Thiol redox and pKa properties of mycothiol, the predomiant low molecular weight thiol cofactor in the Actinomycetes

Herein, the thiol pKa and standard redox potential of mycothiol, the major low molecular weight thiol cofactor in the actinomycetes, are reported. The measured standard redox potential reveals substantial discrepancies in one or more of the other previously measured intracellular parameters that are relevant to mycothiol redox biochemistry

Arbuscular mycorrhizal fungi (AMF) root colonization dynamics of Molinia caerulea (L.) Moench. in grasslands and post-industrial sites

This is an accepted manuscript of an article published by Elsevier in Ecological Engineering on 05/08/2016, available online: https://doi-org.ezproxy.wlv.ac.uk/10.1016/j.ecoleng.2016.06.029 The accepted version of the publication may differ from the final published version.The aims of this studies were: (i) to examine the influence of heavy metal content (Zn, Cd, Pb, Fe, Cu) and other physico-chemical soil parameters on the level of root colonization of Molinia caerulea and (ii) to relate root colonisation parameters and soil variables to Molinia caerulea abundance in two contrasting habitats (grasslands and heavy metal contaminated sites). The sites differ significantly in terms of bio-available heavy metal contents, particularly Zn (34 times more than grasslands), soil texture, CaCO3, organic matter (LOI%), Mg and nitrate content. Principal Component Analysis showed the strong negative correlations between frequency of mycorrhization (F), arbuscular abundance (A%) and intensity of root cortex colonisation (M%) and concentration of bio-available Zn and Cd. Moreover, no positive correlation between root colonization of Molinia and its abundance was found. The frequency of mycorrhization of root fragments (F%) was only slightly different between these two habitats, whereas the intensity of root cortex colonisation (M%) and relative arbuscular abundance (A%) were significantly lower (3 and 4 times respectively) on the post-industrial sites. The bioavailable Zn content in the substratum of post-industrial sites was strongly negatively correlated with species richness, Shannon diversity index and Evenness. In contrast, these relationships were not statistically significant in grasslands. Based on obtained results we could draw a model of possible relationships between root colonization of Molinia, HM content and Molinia abundance on grasslands and post-industrial sites. Bioavailable Zn content in the soil is a one of main factors influencing the Molinia community diversity. In the grasslands, lower amounts of bioavailable Zn, resulted in higher species richness (R) and species diversity (H) which in turn lead to higher root colonization. On the other hand, on the post-industrial sites, the elevated bioavailable Zn content strongly decreases the plant species richness (R) and species diversity (H) and this caused the decline in root colonization parameters. The low species richness on Zn-polluted sites allowed Molinia to reach higher abundance since the competition with other species is reduced

Effect of Long-Term Zinc Pollution on Soil Microbial Community Resistance to Repeated Contamination

The aim of the study was to compare the effects of stress (contamination trials) on the microorganisms in zinc-polluted soil (5,018 mg Zn kg−1 soil dry weight) and unpolluted soil (141 mg Zn kg−1 soil dw), measured as soil respiration rate. In the laboratory, soils were subjected to copper contamination (0, 500, 1,500 and 4,500 mg kg−1 soil dw), and then a bactericide (oxytetracycline) combined with a fungicide (captan) along with glucose (10 mg g−1 soil dw each) were added. There was a highly significant effect of soil type, copper treatment and oxytetracycline/captan treatment. The initial respiration rate of chronically zinc-polluted soil was higher than that of unpolluted soil, but in the copper treatment it showed a greater decline. Microorganisms in copper-treated soil were more susceptible to oxytetracycline/captan contamination. After the successive soil contamination trials the decline of soil respiration was greater in zinc-polluted soil than in unpolluted soil

Horizontal Transfer of a Nitrate Assimilation Gene Cluster and Ecological Transitions in Fungi: A Phylogenetic Study

High affinity nitrate assimilation genes in fungi occur in a cluster (fHANT-AC) that can be coordinately regulated. The clustered genes include nrt2, which codes for a high affinity nitrate transporter; euknr, which codes for nitrate reductase; and NAD(P)H-nir, which codes for nitrite reductase. Homologs of genes in the fHANT-AC occur in other eukaryotes and prokaryotes, but they have only been found clustered in the oomycete Phytophthora (heterokonts). We performed independent and concatenated phylogenetic analyses of homologs of all three genes in the fHANT-AC. Phylogenetic analyses limited to fungal sequences suggest that the fHANT-AC has been transferred horizontally from a basidiomycete (mushrooms and smuts) to an ancestor of the ascomycetous mold Trichoderma reesei. Phylogenetic analyses of sequences from diverse eukaryotes and eubacteria, and cluster structure, are consistent with a hypothesis that the fHANT-AC was assembled in a lineage leading to the oomycetes and was subsequently transferred to the Dikarya (Ascomycota+Basidiomycota), which is a derived fungal clade that includes the vast majority of terrestrial fungi. We propose that the acquisition of high affinity nitrate assimilation contributed to the success of Dikarya on land by allowing exploitation of nitrate in aerobic soils, and the subsequent transfer of a complete assimilation cluster improved the fitness of T. reesei in a new niche. Horizontal transmission of this cluster of functionally integrated genes supports the “selfish operon” hypothesis for maintenance of gene clusters

Interactive and Single Effects of Ectomycorrhiza Formation and Bacillus cereus on Metallothionein MT1 Expression and Phytoextraction of Cd and Zn by Willows

Single and joint ectomycorrhizal (+ Hebeloma mesophaeum) and bacterial (+ Bacillus cereus) inoculations of willows (Salix viminalis) were investigated for their potential and mode of action in the promotion of cadmium (Cd) and zinc (Zn) phytoextraction. Dual fungal and bacterial inoculations promoted the biomass production of willows in contaminated soil. Single inoculations either had no effect on the plant growth or inhibited it. All inoculated willows showed increased concentrations of nutritional elements (N, P, K and Zn) and decreased concentrations of Cd in the shoots. The lowest biomass production and concentration of Cd in the willows (+ B. cereus) were combined with the strongest expression of metallothioneins. It seems that biotic stress from bacterial invasion increased the synthesis of these stress proteins, which responded in decreased Cd concentrations. Contents of Cd and Zn in the stems of willows were combination-specific, but were always increased in dual inoculated plants. In conclusion, single inoculations with former mycorrhiza-associated B. cereus strains decreased the phytoextraction efficiency of willows by causing biotic stress. However, their joint inoculation with an ectomycorrhizal fungus is a very promising method for promoting the phytoextraction of Cd and Zn through combined physiological effects on the plant

Studies on the production of branched-chain alcohols in engineered Ralstonia eutropha

Wild-type Ralstonia eutropha H16 produces polyhydroxybutyrate (PHB) as an intracellular carbon storage material during nutrient stress in the presence of excess carbon. In this study, the excess carbon was redirected in engineered strains from PHB storage to the production of isobutanol and 3-methyl-1-butanol (branched-chain higher alcohols). These branched-chain higher alcohols can directly substitute for fossil-based fuels and be employed within the current infrastructure. Various mutant strains of R. eutropha with isobutyraldehyde dehydrogenase activity, in combination with the overexpression of plasmid-borne, native branched-chain amino acid biosynthesis pathway genes and the overexpression of heterologous ketoisovalerate decarboxylase gene, were employed for the biosynthesis of isobutanol and 3-methyl-1-butanol. Production of these branched-chain alcohols was initiated during nitrogen or phosphorus limitation in the engineered R. eutropha. One mutant strain not only produced over 180 mg/L branched-chain alcohols in flask culture, but also was significantly more tolerant of isobutanol toxicity than wild-type R. eutropha. After the elimination of genes encoding three potential carbon sinks (ilvE, bkdAB, and aceE), the production titer improved to 270 mg/L isobutanol and 40 mg/L 3-methyl-1-butanol. Semicontinuous flask cultivation was utilized to minimize the toxicity caused by isobutanol while supplying cells with sufficient nutrients. Under this semicontinuous flask cultivation, the R. eutropha mutant grew and produced more than 14 g/L branched-chain alcohols over the duration of 50 days. These results demonstrate that R. eutropha carbon flux can be redirected from PHB to branched-chain alcohols and that engineered R. eutropha can be cultivated over prolonged periods of time for product biosynthesis.United States. Dept. of EnergyUnited States. Advanced Research Projects Agency-Energ