毕业论文设计《产漆酶云芝诱变选育》.docVIP

22 - 1 - 产漆酶云芝诱变选育 摘 要 以菌龄为6天的云芝为出发菌株，研究出发菌株的产漆酶活性并制备其原生质体，采用He-Ne激光诱变原生质体，选育云芝高产漆酶菌株。为实践中利用云芝工业化生产漆酶提供了理论依据。 实验结果表明：以浓度为2.06×１０7个／mL 的原生质体进行诱变选育，在最佳诱变条件：激光束波长632.8nm，光斑直径16cm，功率18.70w，距离35cm，照射时间90秒。诱变后的原生质体的再生率为5.76×１０-6，最大产酶量的ΔOD值为222×10-3，比出发菌株的最大产酶量的ΔOD值196×10-3提高了13%。取诱变株接种于斜面连续3次传代测定最大产酶量，结果表明该菌株高产性能具有遗传稳定性。 关键词： 云芝；He-Ne诱变； 原生质体；漆酶 Laccase-PSK protoplast preparation and mutation breeding Abstract To age for six days of the PSK as the starting strain, the strain on the production of paint and the activity of its protoplast, a He-Ne laser induced protoplast, PSK breeding high-yield strains of paint. Practice for the use of industrial production laccase PSK provides a theoretical basis. The results showed that: the concentration of 2.06 ×１０7 / mL for the protoplast mutation breeding, the mutation in the best conditions: a laser beam at 632.8 nm, spot diameter of 16 cm, power 18.70 w, from 35 cm, exposure time 90 seconds . Mutation of the protoplast regeneration rate of 5.76×１０-6, the largest production volume of the Δ OD value of 222×10-3, than the original strain of the largest production volume of Δ OD value of 196×10-3 increase by 13% . Mutation of vaccination for three times in a row in the sloping passage of the largest enzyme production volume, the results show that the performance of high-yield strains of genetic stability. Key words: PSK; He-Ne mutagenesis; protoplast; laccase 目 录 引言5 第一章 材料与方法6 1.1 实验菌种6 1.2 实验主要仪器设备6 1.3 实验药品6 1.4 实验试剂6 1.5 实验培养基7 1.6 菌种的活化8 1.7菌种的扩大培养8 1.8 菌丝体的培养8 1.9出发菌株产漆酶含量检测8 1.10 原生质体的制备、再生8 1.11原生质体制备、He-Ne激光诱变和再生率的测定9 1.12诱变后菌株产漆酶的含量检测10 1.13 稳定性测定10 第二章：结果与分析11 2.1出发菌株的酶量检测11 2.2云芝原生质体制备率和再生率11 2.3原生质体制备、He-Ne激光诱变和再生率的测定12 2.4诱变菌株的产漆酶检测12 2-5-诱变菌株产酶稳定性测试13 结论与展望15 致谢16 参考文献17 附录18 插图清单 图2-112 图2-2... 13 图2-313 图2-4... 14 图2-5... 14 图2-6... 15 表格清单 表2-112 附表119 附表219 附表319 附表420 附表520 附表621 附表721 附表821 附表922 附表1022 附表1123 引 言 1883 年人们首次从漆树漆液中发现漆酶。随后人们发现某些高等真菌也能分泌这种酶, 其中最重要的产生者是担子菌中的白腐菌（White-rot fung