西南师范大学学报(自然科学版)
西南師範大學學報(自然科學版)
서남사범대학학보(자연과학판)
JOURNAL OF SOUTHWEST CHINA NORMAL UNIVERSITY
2014年
2期
48-56
,共9页
王琼%霍影%汪静%张清燕%任尧%葛方兰
王瓊%霍影%汪靜%張清燕%任堯%葛方蘭
왕경%곽영%왕정%장청연%임요%갈방란
胆固醇氧化酶%三相分离%酶动力学方法
膽固醇氧化酶%三相分離%酶動力學方法
담고순양화매%삼상분리%매동역학방법
Cholesterol oxidase%three-phase partitioning%the enzymatic kinetic method
对一株胞外胆固醇氧化酶高产菌 COX48进行了三相分离,结果表明:最优分离条件为:硫酸铵异丙醇沉淀,硫酸铵饱和度为70%,异丙醇与发酵上清液的体积比为0.8∶1时,pH 值为9,在20℃下静置30 min,12000 r/min 离心10 min,纯化倍数4.5倍,回收率高达95%.酶动力学方法检测胆固醇氧化酶测定血清总胆固醇含量,其线性回归方程为 y =0.0064x+0.0027,R 2=0.9654,线性范围0~7.16 mmol/L.结果表明了该酶具有作为临床诊断试剂的潜力,并为临床上利用酶动力学方法测定血清总胆固醇含量奠定了实验基础.
對一株胞外膽固醇氧化酶高產菌 COX48進行瞭三相分離,結果錶明:最優分離條件為:硫痠銨異丙醇沉澱,硫痠銨飽和度為70%,異丙醇與髮酵上清液的體積比為0.8∶1時,pH 值為9,在20℃下靜置30 min,12000 r/min 離心10 min,純化倍數4.5倍,迴收率高達95%.酶動力學方法檢測膽固醇氧化酶測定血清總膽固醇含量,其線性迴歸方程為 y =0.0064x+0.0027,R 2=0.9654,線性範圍0~7.16 mmol/L.結果錶明瞭該酶具有作為臨床診斷試劑的潛力,併為臨床上利用酶動力學方法測定血清總膽固醇含量奠定瞭實驗基礎.
대일주포외담고순양화매고산균 COX48진행료삼상분리,결과표명:최우분리조건위:류산안이병순침정,류산안포화도위70%,이병순여발효상청액적체적비위0.8∶1시,pH 치위9,재20℃하정치30 min,12000 r/min 리심10 min,순화배수4.5배,회수솔고체95%.매동역학방법검측담고순양화매측정혈청총담고순함량,기선성회귀방정위 y =0.0064x+0.0027,R 2=0.9654,선성범위0~7.16 mmol/L.결과표명료해매구유작위림상진단시제적잠력,병위림상상이용매동역학방법측정혈청총담고순함량전정료실험기출.
A strain of extracellular cholesterol oxidase high bacteria COX4-8 by the three phase separation technology has been studied in this paper.Results show that the pH of the supernatant is adjusted to 9.0. Ammonium sulfate (saturation 70%)is added and vortexed gently,followed by addition of isopropanol (80%).The mixture is then incubated at 20 ℃ for 30min for the three-phase formation.These phases are then separated by centrifugation at 12 000 r/min for 10min.The results show that 95% recovery of activi-ty with 4.5 fold purification for cholesterol oxidase has been achieved.