He-Ne Laser Mutagenesis Increasing the Fibrinolytic Enzyme Activity of Bacillus subtilis

doi:10.11924/j.issn.1000-6850.casb20191201001

Abstract

Abstract:

The mutant strain HDBF-N7HN5 was obtained by inducing and selecting Bacillus subtilis strain with high fibrinolytic enzyme. He-Ne Laser mutagenesis was conducted in this study with different mutant time. The mutant strains were selected and the activities of fibrinolytic enzyme were detected with different fibrin plates. The results showed that after 45 min with He-Ne Laser mutagenesis, the fibrinolytic enzyme activity of the mutant strain with the highest yield reached 429.89±5.74 IU/mL. The genetic stability analysis of HDBF-N7HN5 indicated that the production of fibrinolytic enzyme was very stable after 10 generation cultivation. The absolute dissolution rate of blood clot treated with crude fibrinolytic solution reached 57.74±0.72%, and the absolute dissolution rate of blood clot treated with 10-fold dilution could also reach 26.89±0.68%. The fibrinolytic enzyme produced by the strain had good thrombolytic effect in vitro. The results of this study not only increase the microbial fibrinolytic activity, but also provide a basis for the industrialization of fibrinolytic produced by this strain.

Key words: fibrinolytic enzyme, He-Ne laser mutagenesis, mutation breeding, enzymology properties, Bacillus subtilis

CLC Number:

S182

Sun Ying, Wang Haiman, Song Gang, Ge Jingping. He-Ne Laser Mutagenesis Increasing the Fibrinolytic Enzyme Activity of Bacillus subtilis[J]. Chinese Agricultural Science Bulletin, 2020, 36(35): 28-36.

Figures/Tables 16

References 35

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试剂	来源
牛血纤维蛋白	国药品生物制品检定所
牛凝血酶	国药品生物制品检定所
尿激酶原	国药品生物制品检定所
其他试剂	为国产分析纯

试剂	来源
牛血纤维蛋白	国药品生物制品检定所
牛凝血酶	国药品生物制品检定所
尿激酶原	国药品生物制品检定所
其他试剂	为国产分析纯

抗凝时间	PBS(pH 7.8)	粗酶液	10倍稀释粗酶液	尿激酶(256 IU/mL)
0.5 h	—	+	—	—
1 h	—	+	—	—
2 h	—	+	d+	d+
4 h	—	+	d+	d++:

抗凝时间	PBS(pH 7.8)	粗酶液	10倍稀释粗酶液	尿激酶(256 IU/mL)
0.5 h	—	+	—	—
1 h	—	+	—	—
2 h	—	+	d+	d+
4 h	—	+	d+	d++: