DataSheet1_Improving acid resistance of Escherichia coli base on the CfaS-mediated membrane engineering strategy derived from extreme acidophile.docx

Industrial microorganisms used for the production of organic acids often face challenges such as inhibited cell growth and reduced production efficiency due to the accumulation of acidic metabolites. One promising way for improving the acid resistance of microbial cells is to reconstruct their membranes. Herein, the overexpression of cfa2 from extreme acidophile endowed E. coli with high-performance on resistance to the acid stress. The engineered strain M1-93-Accfa2, constructed by CRISPR/Cas9-mediated chromosome integration, also exhibited a significantly higher resistance to severe acid stress. The analysis of fatty acid profiles indicated that the proportion of Cy-19:0 in the cell membrane of M1-93-Accfa2 increased by 5.26 times compared with the control, while the proportion of C18:1w9c decreased by 5.81 times. Correspondingly, the permeability and fluidity of the membrane decreased significantly. HPLC analysis demonstrated that the contents of intracellular glutamic acid, arginine, methionine and aspartic acid of M1-93-Accfa2 were 2.59, 2.04, 22.07 and 2.65 times that of the control after environmental acidification, respectively. Meanwhile, transmission electron microscopy observation indicated that M1-93-Accfa2 could maintain a plumper cell morphology after acid stimulation. M1-93-Accfa2 also exhibited higher-performance on the resistance to organic acids, especially succinic acid stress. These results together demonstrated the great potential of M1-93-Accfa2 constructed here in the production of organic acids.</p

High-strength Al–5Mg₂Si–2Mg–2Fe alloy with extremely high Fe content for green industrial application through additive manufacturing

Achieving superior mechanical properties of Al alloys with high content of Fe impurities is very challenging. Here, a feasible method was applied to accommodate high Fe content (∼2.2 wt.%) and obtain superior strength in an Al–5Mg2Si–2Mg–2Fe alloy by using additive manufacturing. Heterogeneous distribution of Fe, including a high number density of α-Al12(Fe,Mn)3Si particles distributed at the melting pool boundary and excessive Fe segregated along the cell boundaries that divided by Mg2Si eutectics, was verified as the beneficial factor for the alloy design and strength enhancement. In addition to the heterogeneous grains that contain fine cells, the interactions between dislocations and coherent Mg2Si eutectics and the α-Al12(Fe,Mn)3Si particles played an important role in improving the mechanical properties. This work represents a breakthrough in recycling high-strength Al alloys with extremely high Fe doping for green industrial application through additive manufacturing.</p