摘要
对洋葱伯克霍尔德氏菌L 68生长及产邻苯二酚2,3 双加氧酶(C23D)的条件进行了研究,其最适产酶pH7.2;最适生长温度30～35℃;最适培养时间48h;苯酚浓度0.09%最有利于菌体产酶.对菌株L 68产生的C23D酶进行了纯化,超声波破碎后的细胞提取液经硫酸铵分级沉淀、DEAESepharoseFastFlow层析、Hydroxyapatite层析、SephadexG 150层析后,收率为20%,酶比活力提高了230倍.SDS PAGE检测得到了分子量为(34±1)kDa的蛋白.
Burkholderia cepacia L68 can secrete a phenoldegradation enzyme,catechol 2,3dioxygenase(C23D).The enzymeproduction conditions of the bacterium were studied.Strain L68 grew well at neutral pH and the pH optimum for C23D production was 7.2.The optimum temperature for its growth and enzyme production was 30～35 ℃.The specific activity of C23D was the highest when the strain was cultivated for 48 h.The best concentration of phenol was 0.09%.The enzyme was purified by ammonium sulfate precipitation,DEAE Sepharose ionexchange chromatography,Hydroxyapatite Adsorption chromatography,Sephadex G150 chromatography,the recovery rate was 20% and the enzyme activity function after purification was 230 times higher.A single band protein with molecular weight of (34±1) kDa was detected by SDSPAGE.
出处
《生命科学研究》
CAS
CSCD
2003年第2期156-160,共5页
Life Science Research
