基于图像灰度处理的谷氨酰胺转氨酶产生菌半定量-高通量筛选  

Semi-quantitative and high throughput screening of transglutaminase-producing strains based on image grayscale processing

作　　者：石楠 张莹莹[3] 陈海清 李志辉 刘双 于凡 郭璇 檀建新 SHI Nan;ZHANG Yingying;CHEN Haiqing;LI Zhihui;LIU Shuang;YU Fan;GUO Xuan;TAN Jianxin(Engineering Technology Center for Agricultural Products Processing of Hebei Province,College of Food Science and Technology, Hebei Agricultural University,Baoding 071001,China;Key Laboratory of Microbial Diversity Research and Application of Hebei Province,College of Life Sciences,Hebei University,Baoding 071002,China;Chengde Academy of Agriculture and Forestry Sciences,Chengde 067000,China;Office of Logistics Management,Bohai Campus-Hebei Agricultural University,Cangzhou 061100,China)

机构地区：[1]河北农业大学食品科技学院河北省农产品加工工程技术研究中心,河北保定071001 [2]河北大学生命科学学院河北省微生物多样性研究与应用重点实验室,河北保定071002 [3]承德市农林科学院,河北承德067000 [4]河北农业大学渤海校区后勤管理办公室,河北沧州061100

出　　处：《中国酿造》2018年第11期122-126,共5页China Brewing

基　　金：河北省食品科学与工程学科"双一流"建设资金项目(2016SPGCA18);河北省重点研发计划项目(16275505D)

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。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.

关 键 词：拟无枝酸菌109.5 谷氨酰胺转氨酶 图像灰度处理 半定量测定 高通量筛选 

分 类 号：TS201.3[轻工技术与工程—食品科学]