High Pressure-Induced Changes of Physico-Chemical Properties and Mutagenesis in Aspergillus Oryzae
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摘要: 在100~400 MPa压力、保压20 min的条件下处理酱油酿造菌种米曲霉,研究高压对米曲霉存活率、形态特征、生理性质、蛋白酶、淀粉酶活性等的影响,并诱变筛选优良菌株。结果表明:高压对米曲霉的存活率、形态特征有明显的影响;压力对蛋白酶及淀粉酶活性的影响也有特异的规律,即在一定压力范围内(0.1~200 MPa)蛋白酶的活性随着压力的增加而减小,但随着压力的进一步升高(200~400 MPa)蛋白酶的活性又逐渐增强,在300 MPa时超过对照组,400 MPa时蛋白酶的活性达到最高值;淀粉酶在0.1~100 MPa时活性下降,在200 MPa时其平均的糊化和糖化活性最强、活力最高,当压力升高活性又开始降低,400 MPa时几乎又回到对照值。另外,高压处理后获得一株理想的变异菌株HP300a:生长速度快、孢子数量多、蛋白酶活力高,且不易被杂菌污染。其成曲的几项主要指标均优于对照株,酿出酱油的几项主要指标也明显优于对照株。为利用高压诱变筛选米曲霉优良菌种、提高酿造酱油产品的产量及质量提供了理论依据,并发现了高压处理米曲霉引起其蛋白酶及淀粉酶活性改变的特殊规律。Abstract: In order to study the pressure-induced changes of biological and physiochemical properties in fungi, Aspergillus Oryzae was treated with high pressure at 0.1~400 MPa for 20 min, and their viability and modality, as well as the activity of protease and amylase were determined. The result showed that high pressure induced obvious changes in the viability and modality of Aspergillus Oryzae. The activities of protease and amylase in Aspergillus Oryzae showed a specific regular pattern upon high pressure treatment. In the pressure range of 0.1~200 MPa, the protease activities decreased, but enhanced gradually in the range of 200~400 MPa. The protease activity was larger than control strain after treatment fungi with 300 MPa or higher, and reached the maximal at 400 MPa. The activity of amylase also decreased with pressure in the range of 0.1~100 MPa and gradually enhanced in the range of 100~200 MPa, and subsequently decreased again at the range of 200~400 MPa. The activity of amylase is highest in the treatment with 200 MPa, and same as control strain at 400 MPa. A fine mutant strain of fungi, HP300a, was obtained after treatment with 300 MPa. HP300a has a series of excellent characteristic: quick growth, more spores, higher activity of protease, and is rarely polluted by other strains. In addition, several indexes of soy sauce brewaged by HP300a were much better than that by control strains.
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Key words:
- high pressure /
- Aspergillus Oryzae /
- death rate /
- protease activity /
- amylase activity /
- mutagenesis
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Claude B, Ruan K C. An Emerging Field in Bioscience: High Hydrostatic Pressure Study [J]. Acta Biochimica et Biophysica Sinica, 1999, 31(6): 619-624. Kornblatt J A, Kornblatt M J. The Effects of Osmotic and Hydrostatic Pressures on Macromolecular Systems [J]. Biochimica et Biophysica Acta, 2002, 1595: 30-47. Robort B, Macgregor J. The Interactions of Nucleic Acids at Elevated Hydrostatic Pressure [J]. Biochimica et Biophysica Acta, 2002, 1595: 226-276. Takano K J, Takano T, Yamanouchi Y, et al. Pressure-Induced Apoptosis in Human Lymphoblasts [J]. Experiment Cell Research, 1997, 235: 155-160. Shen S L, Wang Z W, Shan X H. Alterations in DNA Methylation and Genomic Structure in High-Pressure Induced Rice Cultivar [J]. Science in China Series C, 2006, 49(2): 97-104. Toshi Otsuki, Masanori Kyo. Deep-Sea Environment Exploration Progam and Deep-Sea Microorganisms Collecting-Cultivating System [J]. High Pressure Science and Technology, 1992, 1(3): 226-233. (in Japanese) 大月 利, 許正憲. 深海環境プログムと深海微生物実験システム [J]. 高圧力の科学と技術, 1992, 1(3): 226-233. Yamazaki Akira. Story of Development of High-Pressure processed Foods, Part 1 [J]. High Pressure Science and Technology, 1997, 8(4): 128-131. (in Japanese) 山崎 彬. 高圧食品開発物語(その1) [J]. 高圧力の科学と技術, 1997, 8(4): 128-131. Yamazaki Akira. Story of Development of High-Pressure processed Foods, Part 2 [J]. High Pressure Science and Technology, 1997, 11(3): 182-186. (in Japanese) 山崎 彬. 高圧食品開発物語(その2) [J]. 高圧力の科学と技術, 1997, 11(3): 182-186. Ludwig H. Cell Biology and High Pressure: Applications and Risks [J]. Biochimica et Biophysica Acta, 2002, 1595: 390-391. Gao X, Li J, Ruan K C. Barotolerant E. Coli Induced by High Hydrostatic Pressure [J]. Acta Biochimica et Biophysica Sinica, 2001, 33(1): 77-81. (in Chinese) 高翔, 李炯, 阮康成. 高压力诱变的耐压大肠杆菌 [J]. 生物化学与生物物理学报, 2001, 33(1): 77-81. Sharma A, Scott J H, Cody G D, et al. Microbial Activity at Gigapascal Pressures [J]. Science, 2002, 295: 1514-1516. Xiang Y, Zhu Z R, Gong S. Investigations of Relationship among Aspergillus Oryza Strains by Isoenzyme Elenzyme Electrophoresis [J]. Chinese High Technology Letters, 1997, (3): 12-16. (in Chinese) 向洋, 朱泽瑞, 龚岁. 米曲霉及激光诱变菌株间的同工酶亲缘关系研究 [J]. 高技术通讯, 1997, (3): 12-16. Xiang Y, Wu J H, Chen S R. Physiological and Biochemical Proterties of Aspergillus Oryzae Strains Radinted by lasers [J]. J Optoelectronics Laser, 1999, 10(2): 153-159. Dang M N, L D J, Fu L. A Screeing Methed of Soy Sauce Aspergillus Orgza [J]. China Condiment, 2004, (9): 17-25. (in Chinese) 党敏娜, 吕东津, 付亮. 一种酱油菌种选筛的方法 [J]. 中国调味品, 2004, (9): 17-25. Wu H Y, Zheng X Q, Yao R. To Obertain High Capacity Carboxypeptidase by Mutagenized with UV Combing LiCl [J]. China Condiment, 2004, (6): 15-17. (in Chinese) 吴红艳, 郑喜群, 姚 蕤. 紫外线-氯化锂合诱变选育羧肽酶高产菌株 [J]. 中国调味品, 2004, (6): 15-17. Li H S, Sun Q, Zhao S J. Principles and Techniques of Plant Physiological Biochemical Experiments [M]. Beijing: Higher Education Press, 2000: 169-172. (in Chinese) 李合生, 孙 群, 赵世杰. 植物生理生化实验原理和技术 [M]. 北京: 高等教育出版社, 2000: 169-172. Meng H H. Shanghai Brewing 3. 042 the Method of Purification and Renovation on Aspergillus Orgza [J]. China Condiment, 2001, (7): 7-9. (in Chinese) 孟颢华. 泸酿3. 042米曲霉的纯化复壮 [J]. 中国调味品, 2001, (7): 7-9. Shen P, Fan X R, Li G W. Microbiology Experiment [M]. Beijing: Higher Education Press, 2002: 120-168. (in Chinese) 沈萍, 范秀容, 李广武. 微生物学实验 [M]. 北京: 高等教育出版社, 2002: 120-168. GB18186-2000. Brewing Soy Sauce [S]. Beijing: China Standard Press, 2001. (in Chinese) GB18186-2000. 酿造酱油 [S]. 北京: 中国标准出版社, 2001.
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