In the genome annotation of Escherichia coli MG1655, the orf382 (1,149 bp) is designated as a gene encoding an alcohol dehydrogenase that may be Fe-dependent. In this study, the gene was amplified from the genome by PCR and overexpressed in Escherichia coli BL21(DE3). The recombinant 6×His-tag protein was then purified and characterized. In an enzymatic assay using different hydroxyl-containing substrates (n-butanol, L-threonine, ethanol, isopropanol, glucose, glycerol, L-serine, lactic acid, citric acid, methanol, or D-threonine), the enzyme showed the highest activity on L-threonine. Characterization of the mutant constructed using gene knockout of the orf382 also implied the function of the enzyme in the metabolism of L-threonine into glycine. Considering the presence of tested substrates in living E. coli cel ls and previous literature, we believed that the suitable nomenclature for the enzyme should be an L-threonine dehydrogenase (LTDH). When using L-threonine as the substrate, the enzyme exhibited the best catalytic performance at 39°C and pH 9.8 with NAD+ as the cofactor. The determination of the Km values towards L-threonine (Km = 11.29 micrometer), ethanol (222.5 micrometer), and n-butanol (8.02 micrometer) also confirmed the enzyme as an LTDH. Furthermore, the LTDH was shown to be an ion-containing protein based on inductively coupled plasma-atomic emission spectrometry with an isoelectronic point of pH 5.4. Moreover, a circular dichroism analysis revealed that the metal ion was structurally and enzymatically essential, as its deprivation remarkably changed the α-helix percentage (from 12.6% to 6.3%).

Eight reinforced concrete beams (six externally reinforced and two contrast beams) were tested under primary or secondary load experiment. According to the test results, the existing specifications were applied to make calculations on short-term deflection which leaded to a large error. The secondary effect should be considered on calculating the deflection. Therefore, comparisons were made between the improved calculated value and test value on deflections of reinforced concrete beams strengthened by externally prestressed transverse tensioning method and they are in good agreement.

In this paper, seven reinforced concrete beams (two contrast beams and five externally reinforced beams) were tested. The crack regularity under different tension stresses was analyzed through experimental data. Taking the stress mode under pressure for the triangle area, in the tension zone in the shape of a trapezoid , the crack resistance equation of externally prestressing reinforced rectangle beam is derived from balance method of the internal forces and moment, and the results coincide with the experiment results well.

Eight reinforced concrete beams (six externally reinforced and two contrast beams) were tested under the primary or secondary load experiment. According to the test results, the existing formulas and specifications were applied to make calculations on reinforced beams. Besides, comparisons were made between test and calculation value, the differences were pointed out and improvement measures were proposed. Comparisons were made between test and calculation value on normal section flexural bearing capacity of reinforced concrete beams strengthened by externally prestressed transverse tensioning method and they are in good agreement.