Enhanced Production of a Thermostable Carbonic Anhydrase in Escherichia coli by Using a Modified NEXT Tag

Carbonic anhydrase (CA) is an ultrafast enzyme that catalyzes the reversible conversion of carbon dioxide (CO2) to bicarbonate. CA is considered to be a green catalyst for enzyme-based CO2 capture and utilization. In particular, the CA of Thermovibrio ammonificans (taCA) has attracted increasing attention as a highly stable enzyme. However, the poor solubility and the low expression level in Escherichia coli have hampered further utilization of taCA. In a recent study, these limitations were partly resolved by using a small solubility-enhancing fusion tag named NEXT, which originates from the N-terminal extension of Hydrogenovibrio marinus CA. In this study, the NEXT tag was engineered by adding small peptides to the N terminus to further increase the production yield of NEXT-tagged taCA. The addition of ng3 peptide (His-Gly-Asn) originating from the N-terminal sequence of Neisseria gonorrhoeae CA improved the expression of NEXT-taCA, while the previously developed translation-enhancing element (TEE) and Ser-Lys-Ile-Lys (SKIK) tag were not effective. The expression test with all 16 codon combinations for the ng3 sequence revealed that the change in translation initiation rate brought about by the change in nucleotide sequence was not the primary determinant for the change in expression level. The modified ng3-NEXT tag may be applied to increase the production yields of various recombinant proteins.

Effect of Milling Parameters on the Structural, Microstructural and Physical Properties of Alumina-Titania-Graphite Nanocomposite

Al2O3-TiO2-graphite nanocomposite can be used as alternative material for coating application. Fine composite coating particles is commonly produced by milling in a high energy ball milling. This study focused on evaluate the effects on the structural, microstructural and physical properties of Al2O3-TiO2-graphite nanocomposite. The alumina, titania and graphite powder were milled in a planetary ball mill at 2, 4, 8 and 10 h and 200, 250 and 300 rpm. The composite particles was compacted for green density determination. The phase analysis and microstructure of nanocomposite were characterized using X-Ray Diffraction (XRD) and Scanning Electron Microscope (SEM). Increasing milling time and milling speed contributes to a small solubility between Al2O3 and TiO2. Increasing milling time and speed decreased the Al2O3 crystallite and internal strain increased as a result of continuous impact on the powder and repeated collision between powder and the wall container. Higher milling time and speed produce finer and flaky shape of Al2O3-TiO2-graphite particles which then affects the green density of the composite.

Titanium Production via Titanium Sulfide

A new metallurgical process via titanium sulfide from ilmenite is proposed and experimentally approved: It consists of several stages; 1) The ilmenite ore is exposed to gaseous CS2 to selectively sulfurize to FeS, which is wet-chemically removed. 2) The residual oxide is again exposed to CS2 to form TiS2. 3) TiS2 is electrochemically reduced to metallic Ti using molten CaCl2-CaS as an application of OS process. TiFeO3 was exposed to Ar-CS2 mixed gas flow at 1173 K to form the mixture of FeS+TiO2. FeS was easily separated by immersing in H2SO4 solution at 313 K. After recovery of TiO2, it was converted completely to TiS2 by the second sulfurization with CS2. TiS2 could be reduced to Ti powder by calciothermic reduction and simulteneous electrolysis in a CaS-CaCl2 melt for about 6 hours at 1173 K and 3.0 V. The impurity decreased to a low level such as 0.021 mass%S due to very small solubility of S in a-Ti. However, 1.06 mass%O remained because of wide solubility of oxygen in a-Ti and water contamination in initial CaCl2.

Oxidation of Four NiAl-Ag Alloys at 900°C in 1 Atm O2

Small amounts of silver have been added to the intermetallic compound β-NiAl for the purpose of improving its mechanical properties. Four ternary NiAl-Ag alloys NiAl-0.5Ag, NiAl-1Ag, NiAl-5Ag and NiAl-10Ag (at.%), and an Ag-free β-NiAl have been oxidized at 900oC for 24 h in 1 atm O2 to study the effect of the presence of silver on the oxidation of β-NiAl. The kinetics of all the alloys were generally composed of two main parabolic stages with slightly larger parabolic rate constants for the second stage, except for NiAl-10Ag, which has an instantaneous parabolic rate constant decreasing with time. A continuous external layer of Al2O3 formed on all the alloys. In particular, the scales formed on NiAl-5Ag and NiAl-10Ag contained a thin and discontinuous layer of silver at the alloy/Al2O3 interface. Furthermore, NiAl-10Ag formed also isolated Ag particles or even a discontinuous Ag layer occasionally surmounting the Al2O3 scale. The addition of minor amounts of silver does not affect significantly the oxidation of β-NiAl, because silver is essentially present as a second phase due to its very small solubility in this intermetallic compound.