Screening for Streptomyces Hygroscopicus Strains with High Production of Agricultural Antibiotics by Streptomycin Resistance

[Objective] The research aimed to screen Streptomyces hygroscopicus strains with high production of agricultural antibiotics. [ Method] A strain of S. hygroscopicus was screened from the soil of Hainan Island. After natural screening and consecutive ultraviolet induced mutation twice, S6-7 strain was obtained as the original strain then treated by UV irradiation and streptomycin resistance screening, and finally rescreened through shake-flask fermentation. [Result] 7 better strains were selected by primary screening from 62 single colonies which were picked out randomly. After 3 generations of consecutive cultivation on slant media and rescreening, 5 strains presented obvious forward mutation. The forward mutation rate reached 8.06%, and the largest production increasing rate came up to 25.11%. [Conclusion] By combining streptomycin resistance screening and conventional ultraviolet induced mutation, both the antibiotic-producing capacity and forward mutation screening efficiency of the original strain were greatly enhanced.

Production of Validamycins from Crude Substrates by Streptomyces hygroscopicus in an External-loop Airlift Bioreactor with a Low Height-to-Diameter Ratio

Fermentation experiments to produce validamycins from crude substrates by Streptomyces hygroscopicus were carried out in an external-loop airlift bioreactor (0.0115 m^3 ) with a low ratio of height to diameter of the riser of 2.9 and a ratio of riser to downcomer diameter of 6.6. The influences of gas flow rate and liquid volume on fermentation of validamycins were investigated. Comparisons of validamycin fermentation were made among the external-loop airlift bioreactor, a mechanically stirred tank bioreactor (0.010m^3 ) and shaking flasks. Under the same operation conditions including fermentation medium composition, inoculum ratio and culture temperature, the fermentation time in the external-loop airlift bioreactor (45 h) was shorter than that in the shaking flasks (100 h) and the same as that in the mechanically stirred tank bioreactor. After a total fermentation time of 45 h under optimized operation conditions, average validamycin concentration obtained in the external-loop airlift bioreactor was close to 19630 μg·ml^-1 validamycin concentration in the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the mechanically stirred tank bioreactor in production of validamycins from crude substrates with dregs by Streptomyces hygroscopicus.

STUDIES ON GLUTAMINE SYNTHETASE FROM Streptomyces hygroscopicus var. jinggangensis

Streptomyces hygroscopicus var. jinggangensis produces validamycin, an important antibiotic used in agriculture. It was found that thexe was a positive correlation between the specific activity of mycelial glutamine synthetase (GS) and validamycin biosynthesis in our laboratory. So, in this paper, the purification, characteristics and regulatory properties of GS are reported. The native enzyme had a molecular weight of approximately 530,000 and was composed of 12 identical subunits, each 43,000. Electronic microscopic examination of preparations negatively stained disclosed that the subunits were arranged in two hexagonal rings that lay one on top of the other in a face-to-face fashion. Mg^(2+) or Mn^(2+) was absolutely needed as cofactor for GS activity. The enzyme activity was regulated by feedback inhibition and cumulative feedback inhibition. In addition, it was also regulated through a covalent modification, adenylylation and deadenylylation, suggesting that the covalent modification of GS exists not only in Gram-negative bacteria but also in some Gram-positive bacteria.