Production Glycerophosphate Oxidase Strains Screening, Identification And Purification And Nature Study

L-a-Glycerophosphate oxidase( GPO, E.C.1.1.3.21) is one of key enzymes to assay the content of triglyceride in the method of enzyme linked immunosorbent assay. With its higher sensitivity and specificity, it has been applied to clinical diagnosis of heart attack and hyperlipemia.Any kinds of GPO from different microorganisms differ from each other in the principle of action and characterization, so as their nutrition and fermentative conditions. In order to know much about its enzymological characterizations, (1)the strain which could produce L-a-Glycerophosphate oxidase was screened; (2)optimization fermentation conditions of producing L-a-Glycerophosphate oxidase form Enterococcus sp. were investigated (3)L-a-Glycerophosphate oxidase from Enterococcus sp. was isolated, purified and its properties were studied. Major results were as follows:1 A strain, Enterococcus sp., which could produce L-a-Glycerophosphate oxidase, was screened from 23 strains. L-a-Glycerophosphate oxidase activity was 0.231 U/mL.2 The liquid fermentative conditions of Enterococcus sp. were studied through single factor and orthogonal design experiment.The best medium for L-a-Glycerophosphate oxidase(GPO) formation was composed of glucose 0.1%, proteose-peptone 1%, yeast extract 1%, K₂HPO₄ 0.5%, glycerol 0.2%, Salt solution 0.25mL, the initial pH was 6.8 .Furthermore, some research about the conditions of fermentation was done. When the germ grew in 250 mL flask containing 100 mL of the medium on the rotary shaker (200r/min) at 30℃ for 18h, GPO had maximal activity, which was 33.8 times than original production. During the course of scale-up culture, keeping other culture conditions, culture circle was 6h shorter than the one of flask fermentation,...