利用高能混合粒子场诱变选育高产Monacolin K、低产桔霉素的红曲霉菌株
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摘要
利用北京正负电子对撞机直线加速器E2束流打靶产生高能混合粒子场,以64.3 Gy的剂量处理两株紫色红曲霉(Monascus Purpureus)M1和M2,经初筛、复筛后采用HPLC法检测诱变菌株中Monacolin K含量,并进行遗传稳定性实验,从而选育高产Monacolin K、低产桔霉素的突变株。结果表明:经混合粒子场辐照后,紫色红曲霉M1和M2分别筛选出43株和22株正突变株,突变率分别为74.64%和56.72%,其中正突变率分别为60.56%和32.84%。突变株M1-20、M2-4发酵液中Monacolin K含量分别达到了421.69 mg/L和406.68 mg/L,较出发菌株分别提高了142.14%和101.27%,经5次传代培养后产Monacolin K的能力仅下降了4.21%和1.65%,并且其发酵液中桔霉素的含量均在0.01~0.04 mg/L之间,与出发菌株相比,桔霉素含量均明显下降。高能混合粒子场可引起紫色红曲霉菌落形态及色泽发生改变,突变率高,可获得遗传性能稳定的突变株,是一种可应用到微生物诱变育种的新方法。
Using of the Beijing electron-positron collider linear accelerator E2 beam shooting produce high-energy hybrid particle field processed two Monascus Purpureus strains M1 and M2 with the dose of 64.3 Gy,after initial and second screening,using HPLC method to detect the Monacolin K content in the mutant,and genetic stability experiments,thus breeding high Monacolin K and low yield citrinin mutant. The results showed that:after hybrid particle field irradiated,Monascus Purpureus M1 and M2 obtained 43 and 22 high yield Monacolin K mutants,respectively,the mutation rate was 74.64% and 56.72%,respectively,in which the positive mutation rate was 60.56% and 32.84%,respectively. Monacolin K content of M1-20 and M2-4 mutant reached421.69 mg/L and 406.68 mg/L,which improved 142.14% and 101.27% than the original strain,respectively,Monacolin K content of M1-20 and M2-4 mutant decreased 4.21% and 1.65% after subculture for 5 times,respectively,and citrinin content of two mutant were between 0.01 mg/L and 0.04 mg/L,which compared with the original strain,citrinin content were significantly decreased. High-energy hybrid particle field could cause colony morphology and color change of Monascus Purpureus,high mutation rate,the mutant strains of genetic stable performance could be obtained,it was a kind of new method that could be applied in the microbial mutation breeding.
Using of the Beijing electron-positron collider linear accelerator E2 beam shooting produce high-energy hybrid particle field processed two Monascus Purpureus strains M1 and M2 with the dose of 64.3 Gy,after initial and second screening,using HPLC method to detect the Monacolin K content in the mutant,and genetic stability experiments,thus breeding high Monacolin K and low yield citrinin mutant. The results showed that:after hybrid particle field irradiated,Monascus Purpureus M1 and M2 obtained 43 and 22 high yield Monacolin K mutants,respectively,the mutation rate was 74.64% and 56.72%,respectively,in which the positive mutation rate was 60.56% and 32.84%,respectively. Monacolin K content of M1-20 and M2-4 mutant reached421.69 mg/L and 406.68 mg/L,which improved 142.14% and 101.27% than the original strain,respectively,Monacolin K content of M1-20 and M2-4 mutant decreased 4.21% and 1.65% after subculture for 5 times,respectively,and citrinin content of two mutant were between 0.01 mg/L and 0.04 mg/L,which compared with the original strain,citrinin content were significantly decreased. High-energy hybrid particle field could cause colony morphology and color change of Monascus Purpureus,high mutation rate,the mutant strains of genetic stable performance could be obtained,it was a kind of new method that could be applied in the microbial mutation breeding.
引文
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[24]印红,谢申义,章光明,等.利用返回式飞船选育优良红曲霉菌[J].核农学报,2004,18(4):297-299.
[25]胡文效,魏彦锋,蒋锡龙,等.高产Monacolin K红曲霉菌种的诱变选育及液态发酵工艺优化[J].食品工业科技,2013,34(24):296-301.
[26]黄艳,孟丽茹,许赣荣,等.高产色素红曲菌的选育及摇瓶发酵培养基优化[J].中国食品添加剂,2013(6):133-139.
[27]付海平.产Monacolin K红曲霉的筛选及其发酵条件的研究[D].长沙:湖南农业大学,2004.
[28]邱雯雯,任雅琳,陈存社,等.常压室温等离子体诱变筛选高乳糖酶活力酵母的研究[J].中国食品学报,2014,14(2):132-137.
[2]Shi Y C,Pan T M.Beneficial effects of Monascuspurpureus NTU 568-fermented products:a review[J].Applied Microbiology&Biotechnology,2011,90(4):1207-1217.
[3]Mapari S A.Fungal polyketideazaphilone pigments as future natural food colorants[J].Trends in Biotechnology,2010,28(6):300-307.
[4]张锐.红曲霉液态发酵生产红曲色素的工艺优化[J].生物技术通报,2015,31(2):187-195.
[5]Endo A.Monacolin K,a new hypocholesterolemic agent produced by a Monascus species.[J].Journal of Antibiotics,2000,32(8):852-854.
[6]杨欢欢,胡中泽.红曲菌的复合诱变及其固态发酵条件的优化[J].食品科学,2012(11):247-251.
[7]李滔滔,李小龙,张凤琴.高产洛伐他汀红曲菌株的诱变选育[J].食品工业科技,2012,33(24):246-248.
[8]Obata K,Ito M,Ochi R,et al.Pharmacological properties of the postsynaptic inhibition by Purkinje cell axons and the action ofγ-aminobutyric acid on Deitersneurones[J].Experimental Brain Research,1967,4(1):43-57.
[9]何晓娇,陈璨,周月婷,等.高产γ-氨基丁酸低产桔霉素红曲霉(Monascusruber)突变子1047筛选与发酵特性研究[J].微生物学杂志,2014,34(5):38-43.
[10]张圆林,王昌禄,陈勉华,等.高产γ-氨基丁酸的红曲霉菌株筛选及发酵条件优化[J].食品科学技术学报,2014,32(5):35-40.
[11]黄志兵,许杨,张泓,等.红曲菌几种主要次级代谢产物及其生物活性的研究进展[J].食品与发酵工业,2010,36(4):143-148.
[12]王璐.农杆菌介导转化及原生质体融合法选育红曲菌Monacolin K高产株[D].无锡:江南大学,2011.
[13]李晶,马贵民,冯晓明,等.红曲霉发酵产物Monacolin K的研究进展与应用[J].食品工程,2014(2):9-11.
[14]胡延岭,刘录祥,郭会君,等.高能混合粒子场诱发的小麦矮杆突变体的SSR分析[J].核农学报,2008,22(4):399-403.
[15]俞法明,严文潮,毛雪琴,等.利用空间诱变技术进行早籼稻新品种的改良[J].核农学报,2014,28(6):949-954.
[16]董亚琳,张洪伟,代秀云.空间环境诱变育种技术及应用[J].农业与技术,2010,33(3):21-23.
[17]高桐梅,卫双玲,李春明,等.芝麻太空诱变效应及AFLP标记检测[J].华北农学报,2013,28(1):227-233.
[18]黄永相,郭涛,蔡金洋,等.空间环境和60Co-γ辐照对水稻稻米品质的诱变效应[J].核农学报,2013,27(6):709-714.
[19]赵金涛,郭新梅,裴玉贺,等.高能混合粒子场对玉米幼苗生长及抗氧化系统的影响[J].核农学报,2014,28(12):2133-2138.
[20]张文涛,杜久元,白斌.作物空间诱变及其在遗传育种中的应用[J].种子,2014,33(4):48-52.
[21]于新玲,刘录祥,乔利仙,等.高能混合粒子场辐照对花生胚小叶组织培养及植株再生的影响[J].核农学报,2012,26(3):433-438.
[22]刘录祥,韩微波,郭会君,等.高能混合粒子场诱变小麦的细胞学效应研究[J].核农学报,2005,19(5):327-331.
[23]李滔滔.高产洛伐他汀红曲霉菌诱变育种及其在酱油废水中的应用[D].株洲:湖南工业大学,2013.
[24]印红,谢申义,章光明,等.利用返回式飞船选育优良红曲霉菌[J].核农学报,2004,18(4):297-299.
[25]胡文效,魏彦锋,蒋锡龙,等.高产Monacolin K红曲霉菌种的诱变选育及液态发酵工艺优化[J].食品工业科技,2013,34(24):296-301.
[26]黄艳,孟丽茹,许赣荣,等.高产色素红曲菌的选育及摇瓶发酵培养基优化[J].中国食品添加剂,2013(6):133-139.
[27]付海平.产Monacolin K红曲霉的筛选及其发酵条件的研究[D].长沙:湖南农业大学,2004.
[28]邱雯雯,任雅琳,陈存社,等.常压室温等离子体诱变筛选高乳糖酶活力酵母的研究[J].中国食品学报,2014,14(2):132-137.