A 4-α-Glucanotransferase from Thermus thermophilus HB8: Secretory Expression and Characterization.

4-α-glucanotransferase (4GT, EC 2.4.1.25) catalyzes the breakdown of the α-1,4 glycosidic bonds of the starch main chain and forms new α-1,4 glycosidic bonds in the side chain, which is often used to optimize the physical and chemical properties of starch and to improve the quality of starch-based food. However, the low enzyme activity of 4GT limits its production and widespread application. Herein, the 4GT gene encoding 500 amino acids from Thermus thermophilus HB8 was cloned and expressed in Escherichia coli. The purified 4GT exhibited maximum activity at pH 7.0 and 60 °C and had a good stability at pH 6.0-8.0 and 30-60 °C. It was confirmed that 4GT possessed the catalytic function of extending the branch length of potato starch. Furthermore, the 4GT gene was successfully expressed extracellularly in Bacillus subtilis. Then, the enzyme yield of 4GT increased by 4.1 times through screening of different plasmids and hosts. Additionally, the fermentation conditions were optimized to enhance 4GT extracellular enzyme yield. Finally, a recombinant Bacillus subtilis with 299.9 U/mL enzyme yield of 4GT was obtained under the optimized fermentation process. In conclusion, this study provides a valuable reference for characterization and expression of food-grade enzymes.

Optimizing Protein-Glutaminase Expression in Bacillus subtilis.

Protein-glutaminase (PG) is a promising protein deaminase. It only hydrolyzes the side chain amido groups of protein-bound to generate ammonia and protein-L-glutamic acid and does not catalyze any other undesirable changes in protein structures. Deamidation of proteins via PG can influence the solubility, emulsification, foaming, and gelation properties of proteins, which are important properties for some food proteins. Therefore, there is great potential for the application of PG in the food industry. PG is derived from Chryseobacterium proteolyticum (C. proteolyticum); however, wild strains are difficult to industrialize because of their low levels of enzyme production. In this article, we studied different strategies for PG expression in B. subtilis. Results showed that PG produced from C. proteolyticum could be successfully secreted in B. subtilis WB800N, and actively secreted in B. subtilis 168(BS168) or DB403 containing a pro-peptide (pro-PG). The secreted PG from B. subtilis WB800N was inactive unless digested by exogenous proteases, such as trypsin, alkaline protease, and neutral protease. However, active PG was secreted by the self-processing of BS168 and DB403. The specific activity of purified PG reached 20.9 U/mg. PG showed maximum activity at pH 5.5, 55 °C and more than 80% of PG activity was retained within a range of pH 3.5-6.5. When Cbz-Gln-Gly was used as the substrate, PG activity was 31.1 ± 0.9 µM min-1 mg-1. Mg2+, Ca2+, and Zn2+ stabilized and even activated PG activity. These strategies concerning PG expression in B. subtilis and the enzymatic properties of PG provide efficient alternatives for PG research and contribute to the industrial-scale production of PG.

SSZ-70 borosilicate delamination without sonication: effect of framework topology on olefin epoxidation catalysis.

We report a scalable delamination procedure for a SSZ-70-framework layered-zeolite precursor, which for the first time does not involve either sonication or long-chain surfactants. Our approach instead relies on the mild heating of layered zeolite precursor B-SSZ-70(P) in an aqueous solution containing Zn(NO3)2 and tetrabutylammonium fluoride. Powder X-ray diffraction data are consistent with a loss of long-range order along the z-direction, while 29Si MAS NMR spectroscopy demonstrates preservation of the zeolite framework crystallinity during delamination. The resulting delaminated material, DZ-2, possesses 1.4-fold higher external surface area relative to the nondelaminated three-dimensional zeolite B-SSZ-70, based on N2 physisorption data at 77 K. DZ-2 was functionalized with cationic Ti heteroatoms to synthesize Ti-DZ-2 via exchange with framework B. Ti-DZ-2 contains isolated titanium centers in its crystalline framework, as shown by UV-Vis spectroscopy. The generality of the synthetic delamination approach and catalyst synthesis is demonstrated with the synthesis of delaminated material DZ-3, which is derived from layered zeolite precursor ERB-1(P) with MWW framework topology. Upon catalytic testing for the epoxidation of 1-octene with ethylbenzene hydroperoxide as oxidant, under harsh tail-end conditions that deactivate amorphous Ti-silica-based catalysts, Ti-DZ-2 exhibits the highest per-Ti-site activity, selectivity, and stability for 1-octene epoxidation of all catalysts investigated. This testing includes the prior benchmark delaminated zeolite catalyst in this area, Ti-UCB-4, which possesses similar external surface area to Ti-DZ-2 but requires sonication and long-chain surfactants for its synthesis. The synthesis of DZ-2 is the first example of an economical delamination of layered zeolite precursor SSZ-70(P) and opens up new doors to the development of delaminated zeolites as commercial catalysts.

A Review of Spatial Variation of Inorganic Nitrogen (N) Wet Deposition in China.

Atmospheric nitrogen (N) deposition (Ndep), an important component of the global N cycle, has increased sharply in recent decades in China. Although there were already some studies on Ndep on a national scale, there were some gaps on the magnitude and the spatial patterns of Ndep. In this study, a national-scale Ndep pattern was constructed based on 139 published papers from 2003 to 2014 and the effects of precipitation (P), energy consumption (E) and N fertilizer use (FN) on spatial patterns of Ndep were analyzed. The wet deposition flux of NH4(+)-N, NO3(-)-N and total Ndep was 6.83, 5.35 and 12.18 kg ha(-1) a(-1), respectively. Ndep exhibited a decreasing gradient from southeast to northwest of China. Through accuracy assessment of the spatial Ndep distribution and comparisons with other studies, the spatial Ndep distribution by Lu and Tian and this study both gained high accuracy. A strong exponential function was found between P and Ndep, FN and Ndep and E and Ndep, and P and FN had higher contribution than E on the spatial variation of Ndep. Fossil fuel combustion was the main contributor for NO3(-)-N (86.0%) and biomass burning contributed 5.4% on the deposition of NO3(-)-N. The ion of NH4(+) was mainly from agricultural activities (85.9%) and fossil fuel combustion (6.0%). Overall, Ndep in China might be considerably affected by the high emissions of NOx and NH3 from fossil fuel combustion and agricultural activities.

Impact of Soil Heavy Metal Pollution on Food Safety in China.

Food safety is a major concern for the Chinese public. This study collected 465 published papers on heavy metal pollution rates (the ratio of the samples exceeding the Grade II limits for Chinese soils, the Soil Environmental Quality Standard-1995) in farmland soil throughout China. The results showed that Cd had the highest pollution rate of 7.75%, followed by Hg, Cu, Ni and Zn, Pb and Cr had the lowest pollution rates at lower than 1%. The total pollution rate in Chinese farmland soil was 10.18%, mainly from Cd, Hg, Cu, and Ni. The human activities of mining and smelting, industry, irrigation by sewage, urban development, and fertilizer application released certain amounts of heavy metals into soil, which resulted in the farmland soil being polluted. Considering the spatial variations of grain production, about 13.86% of grain production was affected due to the heavy metal pollution in farmland soil. These results many provide valuable information for agricultural soil management and protection in China.

Selective molecular recognition by nanoscale environments in a supported iridium cluster catalyst.

The active sites of enzymes are contained within nanoscale environments that exhibit exquisite levels of specificity to particular molecules. The development of such nanoscale environments on synthetic surfaces, which would be capable of discriminating between molecules that would nominally bind in a similar way to the surface, could be of use in nanosensing, selective catalysis and gas separation. However, mimicking such subtle behaviour, even crudely, with a synthetic system remains a significant challenge. Here, we show that the reactive sites on the surface of a tetrairidium cluster can be controlled by using three calixarene-phosphine ligands to create a selective nanoscale environment at the metal surface. Each ligand is 1.4 nm in length and envelopes the cluster core in a manner that discriminates between the reactivities of the basal-plane and apical iridium atoms. CO ligands are initially present on the clusters and can be selectively removed from the basal-plane sites by thermal dissociation and from the apical sites by reactive decarbonylation with the bulky reactant trimethylamine-N-oxide. Both steps lead to the creation of metal sites that can bind CO molecules, but only the reactive decarbonylation step creates vacancies that are also able to bond to ethylene, and catalyse its hydrogenation.

Novel surfactant-free route to delaminated all-silica and titanosilicate zeolites derived from a layered borosilicate MWW precursor.

Layered borosilicate zeolite precursor ERB-1P (Si/B = 11) is delaminated via simultaneous deboronation and SDA removal, to yield material DZ-1 consisting of silanol nests, using a simple aqueous Zn(NO3)2 treatment. Characterization of this synthesis process by PXRD shows loss of long-range order, and transmission electron microscopy (TEM) demonstrates transformation of rectilinear layers in the layered zeolite precursor to single and curved layers in the delaminated material. N2 physisorption confirms the expected decrease of micropore volume and increase in external surface area for delaminated materials relative to their calcined 3D zeolite counterpart. Elemental analysis shows loss of B and absence of Zn in the delaminated material. Resonances corresponding to silanol nests are evident via (29)Si solid-state NMR spectroscopy in DZ-1, which should be located within 12-MR pockets near the external surface. We have successfully utilized these nests as tetrahedral recognition sites for incorporation of Ti within an isolated framework coordination environment in material Ti-DZ-1. Diffuse-reflectance ultraviolet (DR-UV) spectroscopy of Ti-DZ-1 confirms isolated framework Ti sites, which are assigned to bands in the range of 210 nm-230 nm. Infrared spectra of Ti-DZ-1 consist of a distinct absorption band at 960 cm(-1), which is absent in DZ-1 prior to Ti incorporation and has been previously correlated with the presence of framework Ti species. Infrared spectra after pyridine adsorption demonstrate bands consistent with Lewis-acid sites in the resulting Ti-substituted delaminated zeolite. The accessibility of these Lewis-acid sites is confirmed when using Ti-DZ-1 as a catalyst for cyclohexene epoxidation using tert-butyl hydroperoxide as the organic oxidant - a reaction for which both DZ-1 and TS-1 are inactive.

Single-step delamination of a MWW borosilicate layered zeolite precursor under mild conditions without surfactant and sonication.

Layered borosilicate zeolite precursor ERB-1P (Si/B = 11) is delaminated via isomorphous substitution of Al for B using a simple aqueous Al(NO3)3 treatment. Characterization by PXRD shows loss of long-range order, and TEM demonstrates transformation of rectilinear layers in the precursor to single and curved layers in the delaminated material. N2 physisorption and base titration confirm the expected decrease of micropore volume and increase in external surface area for delaminated materials relative to their calcined 3D zeolite counterpart, whereas FTIR and multinuclear NMR spectroscopies demonstrate synthesis of Brønsted acid sites upon delamination. Comparative synthetic studies demonstrate that this new delamination method requires (i) a borosilicate layered zeolite precursor, in which boron atoms can be isomorphously substituted by aluminum, (ii) neutral amine pore fillers instead of rigid and large quaternary amine SDAs, and (iii) careful temperature control, with the preferred temperature window being around 135 °C for ERB-1P delamination. Acylation of 2-methoxynaphthalene was used as a model reaction to investigate the catalytic benefits of delamination. A partially dealuminated delaminated material displays a 2.3-fold enhancement in its initial rate of catalysis relative to the 3D calcined material, which is nearly equal to its 2.5-fold measured increase in external surface area. This simple, surfactant- and sonication-free, mild delamination method is expected to find broad implementation for the synthesis of delaminated zeolite catalysts.

Pd2+/Pd0 redox cycling in hexagonal YMn(0.5)Fe(0.5)O3: implications for catalysis by PGM-substituted complex oxides.

Complex oxides--containing at least two different cations on crystallographically distinct sites--have recently been shown to display redox cycling of platinum group metals (PGMs), such as Pd; for example, Pd-substituted complex oxides can reversibly extrude metallic Pd under reducing conditions and then reincorporate Pd(2+) ions into the lattice under oxidizing conditions. The title compounds, YMn(0.5)Fe(0.5-x)Pd(x)O(3-δ) (0 ≤ x ≤ 0.07) crystallizing in the noncentrosymmetric YMnO(3) structure, were prepared using a sol-gel process at 800 °C, and the structures were refined from high-resolution synchrotron X-ray powder diffraction data. Their redox cycling behavior was monitored using synchrotron X-ray diffraction and EXAFS studies. In contrast to the previously studied complex oxide host compounds, YMn(0.5)Fe(0.5-x)Pd(x)O(3-δ) is only modestly tolerant to cycling: repeated redox cycling leads to the formation of PdO, which, on the time-scale of the oxidation cycles, does not reincorporate in the complex oxide lattice. Both oxidized and reduced samples were tested for the oxidation of CO to CO(2) under CO-lean conditions. YMn(0.5)Fe(0.5-x)Pd(x)O(3-δ) performs essentially as well as previously studied YFe(1-x)Pd(x)O(3-δ). The CO oxidation light-off characteristics of the hexagonal hosts are very similar to finely dispersed PdO. Despite evidence that Pd is almost fully dispersed as divalent ions in the host lattice, which is presumably accompanied by the concurrent creation of oxygen vacancies (2 Pd(2+):1 V(O(2-))), the as-prepared hexagonal materials do not display any significant improvement in catalytic activity as a function of Pd substitution level. This suggests that the corner-connected trigonal bipyramids that characterize this structural family do not enable the transport of oxygen through the bulk of the lattice. The study casts light on factors in the solid-state chemistry of precious metal-substituted complex oxides that influence the efficacy of redox cycling of the precious metal, and catalytic performance.

La4LiAuO8 and La2BaPdO5: comparing two highly stable d8 square-planar oxides.

La(4)LiAuO(8) and La(2)BaPdO(5), two previously known oxides, are presented as model compounds for examining the role of isolated and immobilized Au(3+) and Pd(2+) ions in heterogeneous catalysis. Structural characterization, stability, surface composition, and electronic structure of these compounds are presented. These are examined in studies ranging from synchrotron X-ray scattering, including pair distribution function (PDF) and maximum entropy method (MEM) analysis, to density functional calculations of the electronic structures. The exceptional stability displayed by these compounds as verified by thermogravimetric analysis can be attributed to the presence of covalent Au-O and Pd-O interactions revealed in MEM studies, which suggests a criterion for stabilizing these highly oxophobic transition metals in oxide environments. Catalytic testing of the two compounds as heterogeneous catalysts in the oxidation of CO to CO(2) are presented. La(2)BaPdO(5) appears to be an effective catalyst for CO oxidation, despite the low surface area of the oxide being used. This is the first time that a fully ordered (rather than doped) Pd(2+) oxide had been used to catalyze CO oxidation. La(4)LiAuO(8) on the other hand, is much less effective at catalyzing CO oxidation. Differences in the reactivities of the two compounds are discussed with respect to differences in their density functional electronic structures.

Errors analysis on temperature and emissivity determination from hyperspectral thermal infrared data.

Sensitivity analysis of temperature-emissivity separation method commonly applied to hyperspectral data to various sources of errors is performed in this paper. In terms of resulting errors in the process of retrieving surface temperature, results show that: (1) Satisfactory results can be obtained for heterogeneous land surfaces and retrieval error of surface temperature is small enough to be neglected for all atmospheric conditions. (2) Separation of atmospheric downwelling radiance from at-ground radiance is not very sensitive to the uncertainty of column water vapor (WV) in the atmosphere. The errors in land surface temperature retrievals from at-ground radiance with the DRRI method due to the uncertainty in atmospheric downwelling radiance vary from -0.2 to 0.6K if the uncertainty of WV is within 50% of the actual WV; (3) Impact of the errors generated by the poor atmospheric corrections is significant, implying that a well-done atmospheric correction is indeed required to obtain accurate at-ground radiance from at-satellite radiance for successful separation of land-surface temperature and emissivity.