Directional Breeding of High Itaconic Acid Yielding Strain of <i>Aspergillus terreus</i>with a New Plate Technique

Itaconic acid is commercially produced by the cultivation of Aspergillus terreus using starch hydrolysate as carbon source. The degree of hydrolysis had a great influence on itaconic acid production which was suitable when corn starch was saccharified at 35 DE. The α-amylase was sufficient to drive the starch hydrolysis to the degree. The agar plate assay with LiCl treatment provided a rapid, simple and unequivocal method for screening large numbers of colonies for itaconic acid producing strains. It was learned by experience that the strains on the plates with thick hyphae and light-colored spores often accompanied high itaconic acid production. A strain, designated Ast165, producing itaconic acid with a high yield, was successfully obtained by directional breeding of metabolic end products resistant strains. The itaconic acid concentration produced by Ast165 was 53.8 g/l from 100 g/l of starch hydrolysate in shake flasks. The conversion rate was 61.3%, which was the highest value found in tests.

Immune reaction characteristics and the mechanism of anergy induced by recombinant enterotoxin a of Staphylococcus aureus

To study immune reactions and the mechanism of anergy induced by recombinant enterotoxin A (rSEA) of Staphylococcus aureus. The gene encoding SEA was cloned from standard strain of S. aureus and high efficiently expressed in E. coli. After immunization with purified rSEA, mice were examined for production of specific antibody, subtype of IgG, cytokine mRNA levels such as IFN-γ, IL-2 secretion and T-cell surface PD-1 expression. Results showed that high levels of specific antibodies were produced in two weeks of primary immunization shot. During this time, humoral immune reactions prevailed (IgG2a/ IgG1 【1). During the early phase, Th1 type cytokine mRNA is expressed at a higher level than Th2 type, indicating cellular immune reaction prevailed. Splen- ocyte IFN-γ secretion was significantly decreased after boosting immunization. The PD-1 expression was detected by a flow cytometry examination in the surface of T- lymphocytes which were induced by rSEA, and the expression of PD-1 molecules increased along with the number of boosting and the time after immunization.

Distinct roles of alternative oxidase pathway during the greening process of etiolated algae

The vital function of mitochondrial alternative oxidase(AOX) pathway in optimizing photosynthesis during plant de-etiolation has been well recognized. However, whether and how AOX impacts the chloroplast biogenesis in algal cells remains unclear. In the present study, the role of AOX in regulating the reassembly of chloroplast in algal cells was investigated by treating Auxenochlorella protothecoides with salicylhydroxamic acid(SHAM), the specific inhibitor to AOX, in the heterotrophy to autotrophy transition process. Several lines of evidences including delayed chlorophyll accumulation, lagged reorganization of chloroplast structure, altered PSI/PSII stoichiometry, and declined photosynthetic activities in SHAM treated cells indicated that the impairment in AOX activity dramatically hindered the development of functioning chloroplast in algal cells. Besides, the cellular ROS levels and activities of antioxidant enzymes were increased by SHAM treatment, and the perturbation on the balance of NAD+/NADH and NADP+/NADPH ratios was also observed in A. protothecoides lacking AOX activity, indicating that AOX was essential in promoting ROS scavenging and keeping the redox homeostasis for algal chloroplast development during greening. Overall, our study revealed the essentiality of mitochondrial AOX pathway in sustaining algal photosynthetic performance and provided novel insights into the physiological roles of AOX on the biogenesis of photosynthetic organelle in algae.

Cell responses of Dunaliella salina FACHB 435 (Green Alga) to microgravitational stimulation by clinorotation

Clinorotation experiments were established to simulate microgravity on ground. It was found that there were obvious changes of Dunaliella salina FACHB435 cells and their metabolic characteristics during clinorotation. The changes included the increases of glycerol content, the rate of H + secretion and PM H + ATPase activity, and the decrease of ratio of the plasma membrane (PM) phospholipid to PM protein. These results indicated that microgravity was a stress environment to Dunaliella salina . It is deduced that it would be possible to attribute the effect of microgravity on algal cells to the secondary activation of water stress.