氦氖激光、紫外复合诱变天麻素产生菌华根霉LN-A的原生质体

Mutagenesis on Gastrodin Producing Strain Rhizopus chinensis LN-A by He-Ne Laser and UV Irradiation

摘要:

本文通过研究酶组合、酶浓度、酶作用时间和菌龄等因素对天麻素产生菌华根霉(Rhizopus chinensis) LN-A原生质体制备和再生的影响, 总结出了原生质体制备和再生的最佳条件: 选用对数生长期的菌株, 以蜗牛酶5 mg/mL + 纤维素酶5 mg/mL + 溶菌酶2 mg/mL 30°C保温处理2 h, 原生质体形成率达5.8×107, 原生质体再生率为5.7%。在此基础上首次利用He-Ne激光、紫外线复合诱变天麻素合成菌的原生质体, 当选用15 mW的He-Ne激光辐射原生质体20 min, 再用紫外辐照150 s时获得了转化率及天麻素得率都明显提高的突变株, 其天麻素得率比出发菌株提高20%以上。

Abstract:

The optimum conditions for preparation and regeneration of Rhizopus chinensis LN-A protoplasts were obtained after studying the effects of different factors on them, such as the enzyme composition, concentration and reaction time as well as the different growth phase of the strains. After treating the strains in log phase with concentration of 5 mg/mL snail enzyme, 5 mg/mL cellulose enzyme, 2 mg/mL lysozyme for 2 hours under 30°C, the protoplast formation was up to 5.8×107 and regeneration reached 5.7%. On the basis of this, irradiation of He-Ne laser (λ = 632.8 nm, power = 15 mW) for 20 minutes and UV for 150 seconds were performed on the protoplasts of Rhizopus chinensis LN-A. The mutant LN-B was obtained and the final gastrodin yield was up to 28.9%, which was 20% higher than LN-A.