Cuiyi Liang

The N-terminal hydrophobic sequence of Autographa californica nucleopolyhedrovirus PIF-3 is essential for oral infection

SummaryThe Autographa californica nucleopolyhedrovirus (AcMNPV) open reading frame 115 has been identified as a per os infection factor (pif-3) and is essential for oral infection. Here, we have characterized the pif-3 of AcMNPV in more detail. The pif-3 transcripts were detected from 12 to 96 h post-infection (hpi) in Sf9 cells infected with AcMNPV. Polyclonal antiserum first recognized a 25-kDa protein at 36 hpi. Western blot analysis indicated that PIF-3 is a component of occlusion-derived virus but not of budded virus. The subcellular localization demonstrated that the 21-amino-acid (aa) N-terminal hydrophobic domain of PIF-3, which is conserved in PIF-1, PIF2 and PIF-3, acts as a nuclear location signal and is essential for trafficking the protein to the nucleus. Deletion of either pif-3 or the 21-aa N-terminal hydrophobic domain of pif-3 from AcMNPV abolished per os infectivity but had no effect on the infectivity of the budded virus phenotype.

Optimization of fed-batch enzymatic hydrolysis from alkali-pretreated sugarcane bagasse for high-concentration sugar production.

Fed-batch enzymatic hydrolysis process from alkali-pretreated sugarcane bagasse was investigated to increase solids loading, produce high-concentration fermentable sugar and finally to reduce the cost of the production process. The optimal initial solids loading, feeding time and quantities were examined. The hydrolysis system was initiated with 12% (w/v) solids loading in flasks, where 7% fresh solids were fed consecutively at 6h, 12h, 24h to get a final solids loading of 33%. All the requested cellulase loading (10 FPU/g substrate) was added completely at the beginning of hydrolysis reaction. After 120 h of hydrolysis, the maximal concentrations of cellobiose, glucose and xylose obtained were 9.376 g/L, 129.50 g/L, 56.03 g/L, respectively. The final total glucan conversion rate attained to 60% from this fed-batch process.

Characterization of direct cellulase immobilization with superparamagnetic nanoparticles

Abstract Methods of cellulase immobilization on magnetic particles via glutaraldehyde binding were studied. The binding was confirmed by transmission electronic microscopy (TEM), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) and vibrating sample magnetometry (VSM). Samples analyzed by TEM and XRD showed that the magnetic particles with or without bound cellulase were all nanosized particles with a mean diameter of 11.5 nm, and the binding process did not cause significant changes in particle size and structure. Analysis by FTIR showed that the binding of cellulase to the magnetic nanoparticles might be via covalent bonding between residual amine groups on Fe3O4 nanoparticles and amine groups of the cellulase. The VSM analysis showed that magnetic nanoparticles with or without bound cellulase were all superparamagnetic. The immobilized cellulase had a wider pH and temperature range and improved storage stability compared with the free enzyme. Determination of the Michaelis constants revealed that the immobilized cellulase had a greater affinity for the cellulosic substrate than the free enzyme. The immobilized cellulase showed better performance on hydrolysis of steam-exploded corn stalks than of bleached sulfite bagasse pulp.

A study of CO/syngas bioconversion by Clostridium autoethanogenum with a flexible gas-cultivation system

Bioconversion of CO/syngas to produce ethanol is a novel route in bioethanol production, which can be accomplished by some acetogens. Specific culture vessels and techniques are needed to cultivate these microorganisms since they are anaerobic and substrates are gaseous. In this work, gas-sampling bag was applied as a gas-cultivation system to study CO/syngas bioconversion by Clostridium autoethanogenum and was demonstrated to be efficient because of its flexibility and excellent ability to maintain the headspace atmosphere. C. autoethanogenum can use CO as the sole carbon and energy source to produce ethanol, acetate as well as CO2. In the experimental range, higher ethanol production was favored by higher yeast extract concentrations, and the maximum ethanol concentration of 3.45g/L was obtained at 1.0g/L of yeast extract. Study with various bottled gases showed that C. autoethanogenum preferred to use CO other than CO2 and produced the highest level of ethanol with 100% CO as the substrate. C. autoethanogenum can also utilize biomass-generated syngas (36.2% CO, 23.0% H2, 15.4% CO2, 11.3% N2), but the process proceeded slowly and insufficiently due to the presence of O2 and C2H2. In our study, C. autoethanogenum showed a better performance in the bioconversion of CO to ethanol than Clostridium ljungdahlii, a strain which has been most studied, and for both strains, ethanol production was promoted by supplementing 0.5g/L of acetate.

Production of d-psicose from d-glucose by co-expression of d-psicose 3-epimerase and xylose isomerase

d-Psicose has been drawing increasing attention in recent years because of its medical and health applications. The production of d-psicose from d-glucose requires the co-expression and synergistic action of xylose isomerase and d-psicose 3-epimerase. To co-express these genes, vector pET-28a(+)-dual containing two T7 promoters and RBS sites and an Multiple Cloning Sites was constructed using the Escherichia coli expression plasmid pET-28a(+). The xylose isomerase gene from E. coli MG1665 and the d-psicose 3-epimerase gene from Agrobacterium tumefaciens CGMCC 1.1488 were cloned and co-expressed in E. coli BL21(DE3). After 24h incubation with the dual enzyme system at 40°C, the sugar conversion ratio from d-glucose to d-psicose reached 10%. The optimal conditions were 50°C, pH 7.5 with Co2+ and Mg2+. The d-psicose yields from sugarcane bagasse and microalgae hydrolysate were 1.42 and 1.69g/L, respectively.

Metagenomic analysis for the microbial consortium of anaerobic CO oxidizers

Metagenomics analysis has been applied to identify the dominant anaerobic microbial consortium of the carbon monoxide (CO) oxidizers in anaerobic sludge. Reads from the hypervariable V6 region in the bacterial 16s rDNA were aligned and finally clustered into operational taxonomic units (OTUs). The OTUs from different stages in anaerobic CO condition were classified. Alphaproteobacteria, clostridia, betaproteobacteria and actinobacteria were the most abundant groups, while alphaproteobacteria, betaproteobacteria and actinobacteria were variable groups. CO consumption and production efficiency of the microbial consortium were studied. Semi‐continuous trials showed that these anaerobic CO oxidizers formed a stable microbial community, and the CO conversion rate was at over 84%, with the highest CO consumption activity of 28.9 mmol CO/g VSS●day and methane production activity at 7.6 mmol CH4/g VSS●day during six cycles.