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Abstract

The original strain of Aspergillus oryzae CS3.03 isolated from koji powder was treated with the composite mutagen UV-DES. The mutants were screened with the method of casein plate and agar block. According to re-screening results detecting with the method of Folin-phenol, and the better screening means was found. At the same time, the fermentation ability of high protease activity mutants were tested in the state of low-salt solid-state fermentation technology, and the genetic stability of the best was tested by repeated multiplying. The results showed that the screening efficiency of agar block method was significantly higher than casein plate method. Five mutants of high protease activity with production potential were obtained by re-screening. The result of soy sauce fermentation test proved that the content of ammonia nitrogen and the utilization ratio of total nitrogen were higher than those of the original strain. The ammonia nitrogen in soy sauce fermented by mutant P7 and Y5 increased 14.56% and 11.26% respectively, and the utilization ratio of total nitrogen increased 2.92% and 3.12% in the same situation. Genetic stability test showed that P7 had great industrial application value and genetic stability, and its average protease activity was 4 994.18 U/gdb, which was 173.92% higher than that of the original bacteria.

Publication Date

1-28-2017

First Page

7

Last Page

10,15

DOI

10.13652/j.issn.1003-5788.2017.01.002

References

[1] MULLER C, MCLNTYRE M, HANSEN K, et al. Metabolic engineering of the morphology of Aspergillus oryzae by altering chitin synthesis[J]. Applied and Environmental Microbiology, 2002, 68(4): 1 827-1 836.
[2] 包启安. 酱油科学与酿造技术[M]. 北京: 中国轻工业出版社, 2011: 114.
[3] 邵伟, 熊泽, 唐明. 沪酿3.042米曲霉菌种诱变选育研究[J]. 中国调味品, 2006(3): 21-04.
[4] 凌帅, 刘咏, 姚建铭, 等. 紫外线与N+注入复合诱变选育曲酸高产菌株[J]. 食品科学, 2013, 34(1): 234-238.
[5] 汤斌, 张海龙, 张庆庆, 等. 基于紫外诱变原生质体法选育高活力蛋白酶产生菌[J]. 食品工业科技, 2010, 31(9): 195-200.
[6] ZHENG X F, KOBAYASHI Y, TAKEUCHI M. Construction of a low-serine-type-carboxypeptidase-producing mutant of Aspergillus oryzae by the expression of antisense RNA and its use as a host for heterologous protein secretion[J]. Applied Microbiology and Biotechnology, 1998(49): 39-44.
[7] KITAMOTO N Y, SHOKO Y Y, OHMIYA K I, et a1. A Second Pectin Lyase Gene(pet2) from Aspergillus oryzae KBN616: Its Sequence Analysis and Overexpression, and Characterization of eht Gene Products[J]. Journal of Bioscience and Bioengineering, 2001, 91(4): 378-381.
[8] 陈锡剑, 陈建华. 常压室温等离子体诱变选育高产曲酸米曲霉[J]. 化学与生物工程, 2015, 32(5): 52-60.
[9] 张玉涛, 许学书, 王宏. 不同诱变方法对酱油发酵菌种性能的影响[J]. 食品科学, 2004, 25(4): 39-42.
[10] 周其洋, 陶文沂. 米曲霉多酶系优良菌株的诱变选育[J]. 中国调味品, 2009, 34(9): 57-60.
[11] 蒋雪薇, 盛灿梅, 周倩, 等. 琼脂块法快速平板初筛米根霉L-乳酸高产菌[J]. 食品与机械, 2010, 26(3): 8-10.
[12] 沈萍, 陈向东. 微生物学实验[M]. 4版. 北京: 高等教育出版社, 2007: 242.
[13] 岳希洁, 蒋雪薇, 罗晓明, 等. 酿酒酵母温度敏感性突变株的选育[J]. 食品与机械, 2016, 32(8): 9-12.
[14] 诸葛健, 王正祥. 工业微生物试验技术手册[M]. 北京: 中国轻工业出版社, 1994: 205-209.
[15] 中国国家标准化管理委员会. GB/T 5009.39—2003 酱油卫生分析方法[S]. 北京: 中国标准出版社, 2004.
[16] 苗兰兰, 张东杰, 王颖. 复合诱变高产金属硫蛋白酵母菌株的筛选[J]. 食品科学, 2013, 34(19): 261-264.
[17] 吴华昌, 邓静. 高产蛋白酶米曲霉的选育[J]. 四川食品与发酵, 2004, 40(2): 46-48.

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