海洋低温胆固醇氧化酶生产菌株诱变育种及酶学性质研究
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摘要
利用一株来源于海洋的蜡样芽孢杆菌(Bacillus cereus)XLH059进行紫外诱变、化学诱变及复合诱变,并对诱变菌株的遗传稳定性及胆固醇氧化酶酶学性质进行研究。结果表明,通过复合诱变育种得到酶活力最高、传代后酶活稳定的菌株XL-c23,其所产胆固醇氧化酶的最适pH值为7.0,酶活力在pH 6.0～8.0有较好的稳定性,胆固醇氧化酶最适温度为30℃,30℃以下酶活力较为稳定。
A marine-derived Bacillus cereus XLH059 was mutagenized by ultraviolet mutagenesis, chemical mutagenesis and compound mutagenesis,and the genetic stability and cholesterol oxidase enzymatic properties of the mutagenized strains were studied. The results showed that the strain XL-c23 with the highest enzyme activity and stable enzyme activity was obtained by compound mutation breeding. The optimum pH of the enzyme was7.0, and the enzyme activity was stable at pH 6.0-8.0. The optimum temperature of the enzyme was 30 ℃, and the enzyme activity was stable below 30 ℃.
A marine-derived Bacillus cereus XLH059 was mutagenized by ultraviolet mutagenesis, chemical mutagenesis and compound mutagenesis,and the genetic stability and cholesterol oxidase enzymatic properties of the mutagenized strains were studied. The results showed that the strain XL-c23 with the highest enzyme activity and stable enzyme activity was obtained by compound mutation breeding. The optimum pH of the enzyme was7.0, and the enzyme activity was stable at pH 6.0-8.0. The optimum temperature of the enzyme was 30 ℃, and the enzyme activity was stable below 30 ℃.
引文
[1]曹琦.血脂检测在肺血栓栓塞症中的应用价值[D].济南:山东大学,2018.
[2]邓君明.动植物蛋白源对牙鲆摄食、生长和蛋白质及脂肪代谢的影响[D].青岛:中国海洋大学,2006.
[3]茅益民.肝脏病学药物临床试验[J].中国继续医学教育,2010,2(3):66-74,88.
[4]姚光弼,江绍基.临床肝脏病学进展[J].国外医学(内科学分册),1982(6):261-270.
[5]范建高.2018年《实用肝脏病杂志》精彩回顾[J].实用肝脏病杂志,2019,22(1):1-3.
[6]吴秀秀.尿肾功能及血脂与糖尿病视网膜病变的相关性研究[D].南昌:南昌大学,2018.
[7]肖勇,肖长烨.微生物在不同环境下的进化机制研究进展[J].华侨大学学报(自然科学版),2019,40(1):1-8.
[8]VRIELINK A,GHISLA S.Cholesterol oxidase:biochemistry and structural features[J].FEBS J,2009,276(23):6826-6843.
[9]DOUKYU N.Characteristics and biotechnological applications of microbial cholesterol oxidase[J].Appl Microbiol Biot,2009,83(5):825-837.
[10]PURCELL J P,GREENPLATE J T,JENNINGS M J,et al.Cholesterol oxidase:A potent insecticidal active against boll weevil larvae[J].Biochem Bioph Res Commun,1993,196(3):1406-1413.
[11]VRIELINK A,GHISLA S.Cholesterol oxidase:biochemistry and structural features[J].%FEBS J,2009,276(23):6826-6843.
[12]马礼敦.X射线晶体学的百年辉煌[J].物理学进展,2014,34(2):47-117.
[13]王子天.紫外光/纳米银引起的胁迫刺激下RNR基因调控变化的研究[D].长沙:湖南大学,2014.
[14]李珊.核辐射诱发水稻基因组变异的特征及定向选择技术研究[D].杭州:浙江大学,2016.
[15]刘朝兴.核酸中5-醛基尿嘧啶的合成与检测[D].武汉:武汉大学,2018.
[16]杨震,彭选明,彭伟正.作物诱变育种研究进展[J].激光生物学报,2016,25(4):302-308.
[17]骆行岚.紫外光生电子与顺铂药物协同作用对DNA的损伤及其机理研究[D].福州:福州大学,2014.
[18]季文明,陈毅力,张和春,等.比色法测定胆固醇氧化酶酶活[J].无锡轻工学报,2000,19(3):251-254.
[19]诸葛健.工业微生物实验技术手册[M].北京:中国轻工出版社,1994:35-70.
[20]EI-NAGGAR N E,SOLIMAN H M,El-SHWEIHY N M.Extracellular cholesterol oxidase production by Streptomyces aegyptia,in vitro anticancer activities against rhabdomyosarcoma,breast cancer cell-lines and in vivo apoptosis[J].Sci Rep,2018,8(1):2760.
[21]LASHKARIAN H E.Isolation and identifcation of a native Rhodococcus strain producing cholesterol oxidase from soil[J].Int J Biotechnol Biochem,2010,8(1):205-217.
[2]邓君明.动植物蛋白源对牙鲆摄食、生长和蛋白质及脂肪代谢的影响[D].青岛:中国海洋大学,2006.
[3]茅益民.肝脏病学药物临床试验[J].中国继续医学教育,2010,2(3):66-74,88.
[4]姚光弼,江绍基.临床肝脏病学进展[J].国外医学(内科学分册),1982(6):261-270.
[5]范建高.2018年《实用肝脏病杂志》精彩回顾[J].实用肝脏病杂志,2019,22(1):1-3.
[6]吴秀秀.尿肾功能及血脂与糖尿病视网膜病变的相关性研究[D].南昌:南昌大学,2018.
[7]肖勇,肖长烨.微生物在不同环境下的进化机制研究进展[J].华侨大学学报(自然科学版),2019,40(1):1-8.
[8]VRIELINK A,GHISLA S.Cholesterol oxidase:biochemistry and structural features[J].FEBS J,2009,276(23):6826-6843.
[9]DOUKYU N.Characteristics and biotechnological applications of microbial cholesterol oxidase[J].Appl Microbiol Biot,2009,83(5):825-837.
[10]PURCELL J P,GREENPLATE J T,JENNINGS M J,et al.Cholesterol oxidase:A potent insecticidal active against boll weevil larvae[J].Biochem Bioph Res Commun,1993,196(3):1406-1413.
[11]VRIELINK A,GHISLA S.Cholesterol oxidase:biochemistry and structural features[J].%FEBS J,2009,276(23):6826-6843.
[12]马礼敦.X射线晶体学的百年辉煌[J].物理学进展,2014,34(2):47-117.
[13]王子天.紫外光/纳米银引起的胁迫刺激下RNR基因调控变化的研究[D].长沙:湖南大学,2014.
[14]李珊.核辐射诱发水稻基因组变异的特征及定向选择技术研究[D].杭州:浙江大学,2016.
[15]刘朝兴.核酸中5-醛基尿嘧啶的合成与检测[D].武汉:武汉大学,2018.
[16]杨震,彭选明,彭伟正.作物诱变育种研究进展[J].激光生物学报,2016,25(4):302-308.
[17]骆行岚.紫外光生电子与顺铂药物协同作用对DNA的损伤及其机理研究[D].福州:福州大学,2014.
[18]季文明,陈毅力,张和春,等.比色法测定胆固醇氧化酶酶活[J].无锡轻工学报,2000,19(3):251-254.
[19]诸葛健.工业微生物实验技术手册[M].北京:中国轻工出版社,1994:35-70.
[20]EI-NAGGAR N E,SOLIMAN H M,El-SHWEIHY N M.Extracellular cholesterol oxidase production by Streptomyces aegyptia,in vitro anticancer activities against rhabdomyosarcoma,breast cancer cell-lines and in vivo apoptosis[J].Sci Rep,2018,8(1):2760.
[21]LASHKARIAN H E.Isolation and identifcation of a native Rhodococcus strain producing cholesterol oxidase from soil[J].Int J Biotechnol Biochem,2010,8(1):205-217.