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Effective Microwell Plate-Based Screening Method for Microbes Producing Cellulase and Xylanase and Its Application
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  • Effective Microwell Plate-Based Screening Method for Microbes Producing Cellulase and Xylanase and Its Application
  • Effective Microwell Plate-Based Screening Method for Microbes Producing Cellulase and Xylanase and Its Application
저자명
Kim. Jennifer Jooyoun,Kwon. Young-Kyung,Kim. Ji Hyung,Heo. Soo-Jin,Lee. Youngdeuk,Lee. Su-Jin,Shim. Won-Bo,Jung. Won-Kyo,Hyun. J
간행물명
Journal of microbiology and biotechnology
권/호정보
2014년|24권 11호|pp.1559-1565 (7 pages)
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한국미생물생명공학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Cellulase 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 $100^{circ}C$ 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 min. With this technique, we successfully screened 21 and 31 cellulase- 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.