Effective Microwell Plate-Based Screening Method for Microbes Producing Cellulase and Xylanase and Its Application
- Title
- Effective Microwell Plate-Based Screening Method for Microbes Producing Cellulase and Xylanase and Its Application
- Author
- 현정호
- Keywords
- Bacteria; fungi; screening; cellulose; xylanase; 3,5-dinitrosalicylic acid
- Issue Date
- 2014-11
- Publisher
- 한국미생물·생명공학회
- Citation
- Journal of Microbiology and Biotechnology, v. 24, NO. 11, Page. 1559-1565
- Abstract
- Cellulose and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a 200 mu l total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at 100 degrees C for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 mm. With this technique, we successfully screened 21 and 31 cellulose- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.
- URI
- https://www.jmb.or.kr/journal/view.html?doi=10.4014/jmb.1405.05052https://repository.hanyang.ac.kr/handle/20.500.11754/184353
- ISSN
- 1017-7825;1738-8872
- DOI
- 10.4014/jmb.1405.05052
- Appears in Collections:
- COLLEGE OF SCIENCE AND CONVERGENCE TECHNOLOGY[E](과학기술융합대학) > MARINE SCIENCE AND CONVERGENCE ENGINEERING(해양융합공학과) > Articles
- Files in This Item:
There are no files associated with this item.
- Export
- RIS (EndNote)
- XLS (Excel)
- XML