【摘要】 以图像灰度处理软件Image J为测量手段,以微生物谷氨酰胺转氨酶(MTG)产生菌拟无枝酸菌(Amycolatopsis sp.)109.5为出发菌株,经紫外诱变后,对突变株进行半定量测定和高通量筛选。突变株在含有营养琼脂培养基的96孔板中30℃条件下培养3 d后进行显色反应,利用Image J获得各孔灰度均值,根据灰度均值差异预测突变株的产酶水平。结果表明,从334株突变株中筛选到一株MTG活力为0.803 U/m L的正突变株80,较出发菌株MTG活力提高148.6%。基于Image J图像处理软件与96孔板的固体培养法可以实现MTG产生菌株的半定量测定和高通量筛选。更多还原

【Abstract】 Using the image grayscale processing software Image J as measurement instruments and Amycolatopsis sp. 109.5 with microbial transglutaminase(MTG) production as the starting strain, the mutants was detected by the semi-quantitative and screened by high-throughput after Amycolatopsis sp. 109.5 was induced by ultraviolet(UV). After the mutants were cultured in the 96-well plates containing nutrient agar medium for 3 d at 30 ℃,the color reaction was carried out. The MTG production level of the mutants was predicted according to the differences of the grayscale average obtained by Image J software. The results showed that a positive mutant 80 with the MTG activity of 0.803 U/ml was screened from 334 mutants, which increased 148.6% than that of the starting strain. The semi-quantitative determination and high throughput screening of MTG producing strains could be achieved based on the image grayscale processing software Image J and solid culture method in 96-well plates.更多还原