The title of the article: effects on growth and digestive enzyme activities of the <Emphasis Type="Italic">Hepialus gonggaensis</Emphasis> larvae caused by introducing probiotics

A bacteria strain Hg4-03 of Carnobacterium sp., isolated from the intestine of Hepialus gonggaensis larvae, was fed back to the fourth instars larvae as probiotics to evaluate its impact on the growth performance and digestive enzymes. The larvae were reared in the lab with a natural diet treated with different concentrations of bacterial fermentation and heating killed bacteria, respectively. Compared with the control group, results showed that the growth rates significantly increased and the insect mortality rate decreased significantly after feeding with live probiotics. Meanwhile, the activities of protease, total amylase and trehalase rose significantly in intestinal fluid of the group fed with live probiotics compared with the control treatment. These findings demonstrated that the intestinal bacteria Hg4-03 play an important role for the growth of H. gonggaensis larvae. The bacteria community can improve the growth of H. gonggaensis larvae, indicating that intestinal bacteria may probably be one of the most important factors impacting H. gonggaensis larvae reared in control conditions.     

Enhanced γ-aminobutyric acid-forming activity of recombinant glutamate decarboxylase (gadA) from Escherichia coli

γ-aminobutyric acid (GABA) is an important bioactive regulator, and its biosynthesis is primarily through the α-decarboxylation of glutamate by glutamate decarboxylase (GAD). The procedures to obtain GABA by bioconvertion with high activity recombinant Escherichia coli GAD have been seldom understood. In this study, Escherichia coli GAD (gadA) was highly expressed (about 70–75% of total protein) as soluble protein in Escherichia coli BL21(DE3) containing pET28a-gadA, which was induced by 0.4 mM IPTG in LB medium, and maximal GABA-forming activity of the recombinant GAD was 40 U/mL at a concentration (0.15 mM) of pyridoxal phosphate (PLP) and a concentration (0.6 mM) of Ca²⁺ at optimal pH of 3.8. The optimal concentration (7.5 mM) of Mn²⁺ can also improve the activity of recombinant enzyme, but the co-effect of Ca²⁺ and Mn²⁺ exhibited antagonism effect when added simultaneously. LB and 0.1% (w/v) lactose were selected as culture medium and inducer, respectively. The relative activity was markedly higher activated by Ca²⁺ (174%), Mn²⁺ (164%) than that by other seven bivalent cations. Finally, the yield of GABA was high of 94 g/L detected by paper chromatography or HPLC in 1 L reaction system with 30 mL crude GAD (12 U/mL). By entrapping Escherichia coli glutamate decarboxylase into sodium alginate and carrageenan gel beads, the activity of immobilized GAD (IGAD) remained 85% during the initial five batches and the activity still remained 50% at the tenth batch, these results indicated that the recombinant Escherichia coli GAD was feasible for the future industrial production of GABA.     

A novel xylanase,XynA4-2, from thermoacidophilic <Emphasis Type="Italic">Alicyclobacillus</Emphasis> sp. A4 with potential applications in the brewing industry

A xylanase gene, xynA4-2, was obtained from the genome sequence of thermoacidophilic Alicyclobacillus sp. A4 and expressed in Escherichia coli BL21 (DE3). xynA4-2 encodes a mature protein of 411 residues with a calculated molecular weight of 46.8 kDa. Based on the amino acid sequence similarities (highest identity of 61%), the enzyme was confined into glycoside hydrolase family 10. The purified recombinant XynA4-2 exhibited maximum activity at pH 6.2 and 55°C. The enzyme was stable over a broad pH range, retaining more than 90% of the original activity at pH 5.8–12.0, 37°C for 1 h. The substrate specificity of XynA4-2 was relatively narrow, exhibiting 100, 93, and 35% of the relative activity towards birchwood xylan, oat spelt xylan, and wheat arabinoxylan, respectively. Supplementation of XynA4-2 to mash caused the reduction of mash filtration rate (5.6%) and viscosity (4.0%). When combined with the commercial glucanase from Sunson, higher reduction was detected in the filtration rate (12.0%) and viscosity (17.2%). These favorable properties make XynA4-2 a good candidate in the brewing industry.     

Belowground responses of <Emphasis Type="Italic">Picea asperata</Emphasis> seedlings to warming and nitrogen fertilization in the eastern Tibetan Plateau

The impacts of global climatic change on belowground ecological processes of terrestrial ecosystems are still not clear. We therefore conducted an experiment in the subalpine coniferous forest ecosystem of the eastern edges of the Tibetan Plateau to study roots of Picea asperata seedlings and rhizosphere soil responses to soil warming and nitrogen availability from April 2007 to December 2008. The seedlings were subjected to two levels of temperature (ambient; infrared heater warming) and two nitrogen levels (0 or 25 g m⁻²year⁻¹ N). We used a free air temperature increase from an overhead infrared heater to raise both air and soil temperature by 2.1 and 2.6°C, respectively. The results showed that warming alone significantly increased total biomass, coarse root biomass and fine root biomass of P. asperata seedlings. Both total biomass and fine root biomass were increased, but coarse root biomass was significantly decreased by nitrogen fertilization and warming combined with nitrogen fertilization. Warming induced a prominent increase in soil organic carbon (SOC) and NO₃ ⁻-N of rhizosphere soil, while nitrogen fertilization significantly decreased SOC and NH₄ ⁺-N of rhizosphere soil. The warming, fertilization and warming × N fertilization interaction decreased soil microbial C significantly, but substantially increased soil microbial N. These results suggest that nitrogen deposition combined with warmer temperatures under future climatic change possibly will have no effect on fine root production of P. asperata seedlings, but could enhance the nitrification process of their rhizosphere soils in subalpine coniferous forests.     

Fingerprint-based structure retrieval using electron density

We present a computational approach that can quickly search a large protein structural database to identify structures that fit a given electron density, such as determined by cryo-electron microscopy. We use geometric invariants (fingerprints) constructed using 3D Zernike moments to describe the electron density, and reduce the problem of fitting of the structure to the electron density to simple fingerprint comparison. Using this approach, we are able to screen the entire Protein Data Bank and identify structures that fit two experimental electron densities determined by cryo-electron microscopy.