Production of acid α-Amylase Aspergillus Niger strains by mutagenesis of protoplast
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摘要: 以产酸性α-淀粉酶的黑曲霉0-2-1为出发菌,选用180s的紫外线照射剂量对其孢子原生质体进行诱变,利用固体平板透明圈法初筛和发酵液酶活测定复筛;筛选到3株酶活有较大提高的突变株UV9、UV17、UV31,其酸性α-淀粉酶活力分别为184.66、162.73、167.31U/mL,比出发菌株分别提高了54.9%、38.1%和42.0%,遗传稳定性实验证明3株突变株传代过程中均保持相对稳定的较高酸性α-淀粉酶活力。Abstract: Aspergillus Niger 0-2-1 was used as original strain.This strain of separation and purification of spores protoplast was treated by Ultra Violet ( UV) . The mutant with the bigger ratio of hydrolyzation circle diameter to bacterial colony diameter was screened so as to assay acid α- amylase activities. After genetic stability testing, three strains ( UV9, UV17, UV31) with an high activity of acidα-amylase were screened.Strain UV9, UV17 and UV31 were obtained with an acid α- amylase activity of 184.66, 162.73 and 167.31U /mL.The activities obtained were 54.9%, 38.1% and 42.0% higher than that of the parent strain.
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Keywords:
- Aspergillus Niger /
- acidα-amylase /
- protoplast /
- mutagenesis /
- Ultra Violet (UV
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[1] Ashok P, Poonam N, Carlos RS, et al.Advances in microbial amylases[J].Biochem, 2003, (31) :135-152.
[2] 刘永乐, 李忠海, 杨培华, 等.产耐酸性α-淀粉酶菌株的筛选及其固态发酵条件的优化[J].中国酿造, 2007, 28 (9) :91-93. [3] Yasuji M, Motoo A.Acid-stable-Amylase of Black Aspergilli Part II[J].Agr Biol Chem.1968.32 (1) :104-109.
[4] 李多力, 周培谨.嗜热真菌Thermomyces lanuginosus热稳定α-淀粉酶的纯化及特性[J].微生物学报, 1997, 37 (2) :104-107. [5] 褚清龙, 刘新育, 吕向云, 等.酸性α-淀粉酶高产菌株的原生质体诱变选育[J].安徽农业科学, 2011, 39 (13) :7701-7702. [6] 谢建华, 师永生, 杜丽琴, 等.一株产酸性α-淀粉酶菌株的筛选纯化及酶学性质[J].应用与环境生物学报, 2011, 17 (1) :095-099. [7] Morimura S, Zhang W X, Ichimura T, et al.Genetic Engineering of white Shochu-Koji to achieve Higher Levels of Acid-Stableα-Amylase and Glucoamylase and other Properties when used for Shochu Making on a Laboratory Scale[J].Journal of the Institute of Brewing, 1999, 105 (5) :309-314.
[8] 李文利, 曹泽虹, 董玉玮, 等.黑曲霉原生质体诱变选育内切型菊粉酶生产菌株[J].食品科学, 2012, 33 (09) :144-148. [9] 张丽苹, 徐岩.酸性α-淀粉酶生产菌株的选育的初步研究[J].工业微生物, 2002, 32 (4) :11-14. [10] 陈波, 李大力, 杨树林.酸性α-淀粉酶生产菌株的筛选和酶的纯化及酶的性质研究[J].食品科学, 2005, 26 (5) :119-122. [11] De Bekker C, Wiebenga A, Aguilar G, et al.An enzyme cocktail for efficient protoplast formation in Aspergillus niger[J].Journal of microbiological methods, 2009, 76 (3) :305-306.
[12] 徐良玉, 石贵阳, 陶飞, 等.快速筛选耐酸性α-淀粉酶生产菌株的平板透明圈法[J].无锡轻工大学学报, 2003, 22 (5) :91-94. [13] 李秀珍, 杨平平, 王燕.黑曲霉原生质体诱变育种技术研究进展[J].中国酿造, 2007, 12:1-5. [14] 陈远钊, 罗俊成, 胡佳, 等.酸性α-淀粉酶菌株的诱变及酶的性质研究[J].中国酿造, 2007, 1:10-13. [15] 潘力, 李立风, 彭昶, 等.酱油曲霉孢子原生质体的制备与紫外诱变育种[J].食品与发酵工业, 2006, 32 (8) :1-4. [16] Matsubara T, Ammar Y B, Anindyawati T, et al.Molecular cloning and determination of the nucleotide sequence of raw starch digesting alpha-amylase from Aspergillus awamori KT-11[J].Journal of Biochemistry and Molecular Biology, 2004, 37 (4) :429-438.
[17] 施巧琴, 吴松刚.工业微生物育种学. (第二版) [M].北京:科学出版社, 2003.71-74. [18] 李勃.微生物发酵生产耐酸性α-淀粉酶的研究[D].西安:西北大学, 2009.
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