The Screening Of The Glycosaminoglycan Product Strain And The Optimization Of Its Ferment Condition

Utilize glycosaminoglycan's character of producing white turbidity through reacting with quaternary ammonium salt, for example, cetylpyridinium chloride (CPC), to obtain 4 screened strains from 500 ones of various sources. A glycosaminoglycan product strain GS36 is obtained by agarose gel electrophoresis. The infra-red spectrum tested after the purification of the ferment product is identical with the standard infra-red spectrum of the chondroitin sulfate. Therefore, the product is proved to be chondroitin sulfate. Through the observance of its shape and the physiological and biochemical test, it is identified as B.firmus.Through the single factor test and the orthogonal test, the ferment condition of the strain is further optimized: take 1.5% glucose asthe carbon source, 0.3% beef extract as the nitrogen source, add 0.1% Na₂SO₄, 0.1% K₂HPO₄ and 0.02% MgSO₄, with culture medium starting with PH7. Cultivated for 24h under 30℃ through shaking the bottle, the concentration of the ferment product is 0.133mg/ml.Through the extraction and agarose gel electrophoresis, plasmid is found. After elimination of it, the concentration of the ferment product is largely decreased. Thus it is presumed that the gene which regulates the vitality of enzyme exists in the plasmid.Inoculate the activated strain to the ferment culture medium,cultivating for 24h. Compare intracellular and extra cellular concentration of the product. It is concluded that the synthetic product of the strain mainly exists extracellularly.