Ten Leonid N, Im Wan-Taek, Kim Myung-Kyum, Kang Myung Suk, Lee Sung-Taik
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 371-1 Kuseong-Dong, Yuseong-Gu, Daejeon 305-701, Republic of Korea.
J Microbiol Methods. 2004 Mar;56(3):375-82. doi: 10.1016/j.mimet.2003.11.008.
A plate assay based on the visible solubilization of small substrate particles and the formation of haloes on Petri dishes, containing a mixture of different dye-labelled polysaccharides as substrates, provides a specific, reliable and rapid simultaneous detection of corresponding polysaccharide-degrading microorganisms. It has potential for increasing the efficacy of screening of microorganisms, utilizing different polysaccharides, in large numbers of natural samples. Diversely colored insoluble forms of amylose, xylan and hydroxyethyl-cellulose (HE-cellulose) were prepared as chromogenic substrates by using the cross-linking reagent 1,4-butanediol diglycidyl ether and the dyes Brilliant Red 3B-A, Cibacron Blue 3GA and Reactive Orange 14. Using the method, the bacteria with amylase or xylanase or cellulase or a combination of these activities were screened from soil and sludge samples, selected and identified according to 16S rDNA sequencing.
一种基于小底物颗粒的可见溶解以及在含有不同染料标记多糖混合物作为底物的培养皿上形成晕圈的平板检测方法，可对相应的多糖降解微生物进行特异性、可靠且快速的同时检测。它具有提高利用不同多糖对大量天然样品中的微生物进行筛选效率的潜力。通过使用交联剂1,4 - 丁二醇二缩水甘油醚和染料亮红3B - A、汽巴克隆蓝3GA以及活性橙14，制备了不同颜色的直链淀粉、木聚糖和羟乙基纤维素（HE - 纤维素）不溶性形式作为显色底物。利用该方法，从土壤和污泥样品中筛选出具有淀粉酶或木聚糖酶或纤维素酶或这些酶活性组合的细菌，并根据16S rDNA测序进行选择和鉴定。
