产凝乳酶米黑毛霉的诱变育种研究
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摘要
以产凝乳酶的米黑毛霉为出发菌株,通过紫外线照射、紫外复合氯化锂处理,以筛选产凝乳酶活力较高的菌株。确定的紫外线最佳照射时间为90 s,经多次诱变,筛选得到了突变株UV-13,其凝乳酶活力为2448.98 SU/m L,比出发菌株提高了42.85%;复合诱变的最佳条件为90 s紫外照射复合1.5%氯化锂,在多次诱变的基础上,筛选得到了突变株UV-Li Cl-6,其凝乳酶活力为2703.58 SU/m L,比出发菌株提高了57.70%。传代实验表明,两株突变菌都具有良好的遗传稳定性。
In order to breed high activity milk- clotting enzyme from strain,Mucor miehei was used as the origin strain and mutagenized by Ultraviolet and Ultraviolet combining lithium chloride.The optimal condition of Ultraviolet was 90 s. After screening repeatedly,a mutant strain UV- 13 was screened. The milk- clotting activity of UV- 13 reached 2448.98 SU / m L and was 42.85% higher than that of the parent strain. The best condition of compound mutation were that the time of Ultraviolet was 90 s,the concentration of lithium chloride was 1.5%.On the basis of multiple mutation,a mutant strain UV- Li Cl- 6 was screened. The milk- clotting activity of UV- Li Cl- 6 reached2703.58 SU / m L and was 57.70% higher than that of the parent strain.Results of the pass- generation test indicated that two strains had good genetic stability.
In order to breed high activity milk- clotting enzyme from strain,Mucor miehei was used as the origin strain and mutagenized by Ultraviolet and Ultraviolet combining lithium chloride.The optimal condition of Ultraviolet was 90 s. After screening repeatedly,a mutant strain UV- 13 was screened. The milk- clotting activity of UV- 13 reached 2448.98 SU / m L and was 42.85% higher than that of the parent strain. The best condition of compound mutation were that the time of Ultraviolet was 90 s,the concentration of lithium chloride was 1.5%.On the basis of multiple mutation,a mutant strain UV- Li Cl- 6 was screened. The milk- clotting activity of UV- Li Cl- 6 reached2703.58 SU / m L and was 57.70% higher than that of the parent strain.Results of the pass- generation test indicated that two strains had good genetic stability.
引文
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[15]李平,宛晓春,陶文沂,等.复合诱变黑曲霉选育β-葡萄糖苷酶高产菌株[J].菌物系统,2000,19(1):117-120.
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[21]吴振倡,王凤仙,陈守川,等.铜蒸气激光及其与氯化锂复合选育龟裂链霉菌的研究[J].激光生物学报,1997,6(2):1068-1070.
[2]El-Bendary M A,Maysa E M,Thanaa H A.Purification and Characterization of Milk Clotting Enzyme Produced by Bacillus sphaericus[J].Journal of Applied Sciences Research,2007,3(8):695-699.
[3]Vishwanatha K S,Appurao A G,Singh S A.Production and characterization of a milk-clotting enzyme from Aspergillus oryzae MTCC 5341[J].Applied Microbiology and Biotechnology,2010,85(6):1849-1859.
[4]Seker S,Beyenal H,Ayhan F,et al.Production of Microbial Rennin from Mucor miehei in a continuously fed fermenter[J].Enzyme and Microbial Technology,1998,23(7):469-474.
[5]Garg S K,Johri B N.Rennet:current trends and future research[J].Food Reviews International,1994,10(3):313-355.
[6]Hashem A M.Optimization of milk-clotting enzyme productivity by Penicillium oxalicum[J].Bioresource Technology,1999,70(2):203-207.
[7]潘道东,韩玲玲.根霉凝乳酶的分离纯化及其酶学特性研究[J].中国食品学报,2011,11(2):53-58.
[8]He X L,Ren F Z,Guo H Y,et al.Purification and properties of a milk-clotting enzyme produced by Bacillus amyloliquefaciens[J].Korean Journal of Chemical Engineering,2011,28(1):203-208.
[9]徐伟.微波诱变乳酸高产菌选育及淀粉废水发酵制乳酸工艺研究[D].哈尔滨:哈尔滨工业大学,2009.
[10]孟甜,李玉锋.现代工业微生物育种技术研究进展[J].生命科学仪器,2009,7(9):3-6.
[11]卢燕云,林建国,李明,等.复合诱变选育酸性蛋白酶高产菌株[J].中国酿造,2009(1):49-51.
[12]吴红艳,郑喜群,姚蕤.紫外线-氯化锂复合诱变选育羧肽酶高产菌株[J].中国调味品,2004(6):15-17.
[13]王陶,李文,尹龙,等.紫外-氯化锂复合诱变选育中性植酸酶高产菌株[J].食品科学,2012,33(13):217-220.
[14]刘芳宁,梁琪,乔海军,等.紫外线与氯化锂诱变选育高产乳糖酶米曲霉[J].食品工业科技,2013,34(1):155-159.
[15]李平,宛晓春,陶文沂,等.复合诱变黑曲霉选育β-葡萄糖苷酶高产菌株[J].菌物系统,2000,19(1):117-120.
[16]诸葛健,王正祥.工业微生物实验技术手册[M].北京:中国轻工业出版社.1997.
[17]邵淑娟.产疑乳酶霉菌菌种的诱变选育及其酶学性质研究[D].长春:吉林大学,2011.
[18]Arima K,Iwasake K.Milk-clotting enzyme from microorganisms.Part I Screening test and the identification of the potent fungus[J].Agricultural Biological Chemistry,1967,31(5):540-545.
[19]Kumar S,Sharma N S,Saharan M R,et al.Extracellular acid protease from Rhizopus oryzae:purificationand characterization[J].Process Biochemistry,2005,40(5):1701-1705.
[20]周俊清.凝乳酶优良菌株的选育及其酶活特性的研究[D].长沙:湖南农业大学,2005.
[21]吴振倡,王凤仙,陈守川,等.铜蒸气激光及其与氯化锂复合选育龟裂链霉菌的研究[J].激光生物学报,1997,6(2):1068-1070.