Effective screening of cellulase and xylanase producing microbes by modified DNS method

Abstract

In this study, we screened cellulase and xylanase producing microbes by medium control and 96 microplate-well based 3,5-dinitrosalicylic acid (DNS) modifying method. We controlled medium with seawater and underwater as basic water. First culture was carried out with cellulose and xylan both as substrate on agar plate. After pure culture, each colony was re-cultured in broth with cellulose and xylan substrate, respectively. Cultured supernatants were tested activity with modified 3,5-dinitrosalicylic acid (DNS) method with 200 ul total reaction volume each in 96 microwell-plate. Among total of 116 strains, 19 of them were screened to secrete both type of cellulase and xylanase such as Vibrio, Bacillus, Alternaria, Graphium, Planococcus, Nectria, Trichoderma, Streptomyces, Ochrobactrum, Frondihabitans, Cryptococcus, Cladosporium, and Cellvibrio genera. These are important for bio-ethanol production.lture was carried out with cellulose and xylan both as substrate on agar plate. After pure culture, each colony was re-cultured in broth with cellulose and xylan substrate, respectively. Cultured supernatants were tested activity with modified 3,5-dinitrosalicylic acid (DNS) method with 200 ul total reaction volume each in 96 microwell-plate. Among total of 116 strains, 19 of them were screened to secrete both type of cellulase and xylanase such as Vibrio, Bacillus, Alternaria, Graphium, Planococcus, Nectria, Trichoderma, Streptomyces, Ochrobactrum, Frondihabitans, Cryptococcus, Cladosporium, and Cellvibrio genera. These are important for bio-ethanol production.