黑木耳漆酶高产菌株的筛选

Screening for High Yield Laccase Producing Strains from Auricularia auricula

为筛选黑木耳漆酶高产菌株以便对黑木耳漆酶酶学性质、基因表达及结构与功能进行研究,从河南省农业科学研究院、河南省生物研究所购得黑木耳菌株10株。首先用含有0.04%愈创木酚的PDA固体培养基进行平板初筛,再进行液体培养基摇瓶培养,ABTS为底物检测酶活进行复筛,对筛选出来的菌株进行Native-PAGE,以判断黑木耳漆酶的同工酶数量。结果发现初筛培养基中,变色圈出现的时间早晚有差异,但一段培养时间后的直径大小差异不大;液体培养筛选得到一个黑木耳漆酶高产菌株地茂1号,其在培养第11天时酶活达到最大值(386.85U/L);酶活最高峰时取培养液,离心后的粗酶液进行Native-PAGE,底物染色后发现地茂1号粗酶液中含有3种漆酶同工酶。

To screen the strains with high production of laccase from A. Auricula so that the properties and gene expression of A. Auricula laccase as well as its structures and functions can be studied,10 strains were brought from Henan Agricultural Academy of Science and Henan Institution of Biology, respectively. All strains grown on PDA plate containing 0.04% guaiacol were selected according to their diameters and shades of coloration circle. When further screening was taken in submerged fermentation, Dimao-1 showed highest level of laccase activity at 363.85 U/L after cultured in shaken flasks for 11 days. Dimao-1 was potential strains of A. auticula for further research in laccase gene clone and expression. Three kinds of laccases, were identified by the colomess development of zymogram stained with guaiacol after native polyacrylamide electrophoresis.