Heterologous expression and site-directed mutagenesis of endoglucanase CelA from Clostridium thermocellum

The endoglucanase CelA from Clostridium thermocellum was strongly expressed in Bacillus subtilis. The enzyme was purified by Ni²⁺-affinity chromatography. Site-directed substitution of D278 with an asparagine or an alanine residue surprisingly did not decrease the apparent k _cat value. Further substitutions of two other potentially critical residues, Y215 and D152, resulted in a 2-fold decrease in apparent k _cat value for Y215P and complete loss of activity for D152N.     

The dynamics of microflora and the occurrence of phytotoxic substances in different soils with corn residues and inorganic nitrogen

In an incubation experiment the development dynamics of bacterial and fungal communities as well as the level of phytotoxicity were analysed in sand and three soils differing in mechanical structure and amended with corn residues and mineral nitrogen. Bacterial biomass was positively correlated with the degree of dispersion of the solid phase of the soil, whereas the ratio of fungal to bacterial biomass (F:B) was found to be negatively correlated. Fungi were much more tolerant to carbon or nitrogen deficit than bacteria. Introduction of the plant material alone, characterized by a broad carbon to nitrogen ratio, led to the domination of fungi in microbial communities. The level of soil phytotoxicity built up with increasing level of crop residues. Phytotoxicity was observed for the longest time period in soil with the highest silt and clay content. The narrowing of the C:N ratio at introduction of the appropriate amount of mineral nitrogen (larger in heavier soils) resulted in accelerated disappearance of phytotoxicity and at the same time favoured bacterial development. This points to a significant participation of bacteria in the degradation of phytotoxic substances in the soil.     

Heterotrophic production of Chlorella sp. TISTR 8990—biomass growth and composition under various production conditions

The green microalga Chlorella sp. TISTR 8990 was grown heterotrophically in the dark using various concentrations of a basal glucose medium with a carbon‐to‐nitrogen mass ratio of 29:1. The final biomass concentration and the rate of growth were highest in the fivefold concentrated basal glucose medium (25 g L^?1 glucose, 2.5 g L^?1 KNO₃) in batch operations. Improving oxygen transfer in the culture by increasing the agitation rate and decreasing the culture volume in 500‐mL shake flasks improved growth and glucose utilization. A maximum biomass concentration of nearly 12 g L^?1 was obtained within 4 days at 300 rpm, 30°C, with a glucose utilization of nearly 76% in batch culture. The total fatty acid (TFA) content of the biomass and the TFA productivity were 102 mg g^?1 and 305 mg L^?1 day^?1, respectively. A repeated fed‐batch culture with four cycles of feeding with the fivefold concentrated medium in a 3‐L bioreactor was evaluated for biomass production. The total culture period was 11 days. A maximum biomass concentration of nearly 26 g L^?1 was obtained with a TFA productivity of 223 mg L^?1 day^?1. The final biomass contained (w/w) 13.5% lipids, 20.8% protein and 17.2% starch. Of the fatty acids produced, 52% (w/w) were saturated, 41% were monounsaturated and 7% were polyunsaturated (PUFA). A low content of PUFA in TFA feedstock is required for producing high quality biodiesel. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1589–1600, 2017     

Fate of [carbonyl-14C]methabenzthiazuron in an arid region soil — effect of organic amendment,soil disturbance and fumigation

[Carbonyl-¹⁴C] methabenzthiazuron (MBT) was applied to an arid region soil at a rate of 5mg kg⁻¹ soil to give a¹⁴C content of 2400 KB kg⁻¹ soil. After 15 weeks of incubation at 22°C and 50% of the maximum water holding capacity of the soil, 7.2% of the applied¹⁴C was mineralized to¹⁴CO₂. Where the soil was amended with wheat straw, total mineralization increased to 17.3%. Soil disturbance caused a significant increase while chloroform fumigation caused a significant decrease in the rate of¹⁴CO₂ production, both from amended and unamended soils. These results suggest that MBT is degraded mainly through microbial co-metabolism. Wheat straw amendment resulted in increased transformation of MBT into soil humus. In unamended soil, a major portion of¹⁴C was recovered in fulvic acid and in fractions extracted with organic solvents. Recovery of¹⁴C in non-extractable bound residues (humins) increased as incubation progressed and seemed to be derived from the fulvic acid fraction, which showed a concomitant decrease. More than 99% of the residual¹⁴C in unamended soil consisted of unaltered MBT; the remainder occurred as 1-methyl-1 (benzthiazolyl) urea. In amended soil, a relatively higher percentage of the extractable¹⁴C was found in the metabolite. Small amounts of three unidentified¹⁴C-labelled compounds were also observed. In amended soil, disturbance caused a decrease in extractable-¹⁴C whereas fumigation caused a significant increase, as compared to the untreated control. The effects were more pronounced when the soils were reated at an early stage of incubation. In general, soil disturbance increased the availability of MBT for further transformations while chloroform fumigation decreased the process.     

Should China subsidize cofiring to meet its 2020 bioenergy target? A spatio‐techno‐economic analysis

China has developed ambitious bioenergy installation targets as part of its broader goals to increase its renewable energy‐generating capacity and decarbonize its economy. A key target feedstock for bioenergy is the 800 million tonnes of agricultural residues that China produces each year. At present, the main financial incentive to support bioenergy generation from agricultural residues is a feed‐in‐tariff provided for bioenergy that is produced by units that take 80% or more of their feedstock energy from biomass. Although this policy has catalysed the construction of many bioenergy units, there are reports that these projects are experiencing serious financial and technical problems, leading to low operational efficiency and even closure. An alternative option for China's agricultural residues is cofiring with coal in existing power stations. However, this is currently unprofitable for power station operators, as cofiring is not eligible for financial assistance through the bioenergy feed‐in‐tariff. In the light of China's ambitious target to install 30GW of bioenergy generation capacity by 2020, this study investigates the extent to which extension of the bioenergy feed‐in‐tariff to include cofiring could contribute towards this goal. The results suggest that 39% of China's straw energy resources are located within 50 km of a power station. Assuming cofiring ratios of up to 10% coal energy replacement, an annual 89–117TWh of electricity could be generated by cofiring agricultural residues collected within 50 km radii of power stations. If China extends its bioenergy subsidies to include cofiring, an annual 62–92TWh can be produced at an internal rate of return of 8% or more. This equates to 42–62% of the bioenergy generation that China might expect if it met its 2020 target of installing 30GW of bioenergy capacity. Overall, this indicates a strong case for the Chinese government to extend its existing bioenergy feed‐in‐tariff to include cofiring at low energy replacement ratios.     

纤维素降解菌的分离筛选及对中药废弃物的固相发酵研究

中药在提取炮制后残留大量生物质被简单堆弃,而其中的木质纤维素等生物质能被微生物有效利用降解,筛选到16株对木质纤维素有降解效果的菌株,对其中8株降解效果明显功能菌株进行了深入研究,经16(18)S rDNA鉴定发现5株细菌主要为Bacillus属、Streptomyces属和Enterobacter属,3株真菌为Ascomycete属、Aspergillus属和Trichosporon属;利用8株菌固相发酵中药废弃物,通过监测堆体温度、pH等参数,发现细菌类在发酵初期能迅速提高堆体温度,而真菌类发酵腐熟过程较缓慢,提示利用细菌-真菌联合组成降解菌系是提高中药废弃物固相发酵的重要手段。     

The three-dimensional structure of mitochondrial aspartate aminotransferase at 4.5 A resolution

An X-ray crystallographic study at 4.5 Å resolution has been carried out with triclinic crystals of chicken mitochondrial aspartate aminotransferase.In the electron density map, the enzyme is clearly visible as an isologous α₂-dimer (105 Å × 60 Å × 50 Å) in which the subunits are associated about a molecular 2-fold axis. Each subunit of dimensions 70 Å × 50 Å × 40 Å contains at least seven helices, one of which is about 50 Å long.Difference maps have revealed the positions of the pyridoxyl and the phosphate moieties of the coenzyme as well as the general substrate binding area. The active sites are on opposite sides of the dimer, about 30 Å apart and close to the intersubunit boundary, so that probably both subunits contribute to each active site. An isolated chain segment, passing in front of the active site and ending in contact with the neighbouring subunit is interpreted as one of the chain termini.