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Construction of a Metagenomic Library from Compost and Screening of Cellulase- and Xylanase-positive Clones
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  • Construction of a Metagenomic Library from Compost and Screening of Cellulase- and Xylanase-positive Clones
  • Construction of a Metagenomic Library from Compost and Screening of Cellulase- and Xylanase-positive Clones
저자명
Kwon. Eun-Ju,Jeong. Yu-Seok,Kim. Yong-Ho,Kim. Sung-Kyum,Na. Han-Beur,Kim. Jung-Ho,Yun. Han-Dae,Kim. Hoon
간행물명
Journal of the Korean Society for Applied Biological Chemistry
권/호정보
2010년|53권 6호|pp.702-708 (7 pages)
발행정보
한국응용생명화학회
파일정보
정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
서지반출

기타언어초록

Metagenomic library was constructed from compost made with pig manure and mushroom cultural waste using fosmid vector. Composting was carried out using moving roller system with intermittent mixing and forced aeration, and samples were taken from $7^{th}$- and $14^{th}$-day spots, which represent the mid and late stages of the process, respectively. DNAs of about 40 kb were obtained by preparative electrophoresis on 0.4% low-melting agarose gel to fractionate DNA fragments and to remove humic substances. Total 12,380 fosmid clones were obtained. Restriction analysis of randomly selected clones showed that most clones had different inserts, and average size of inserts was about 35 kb. Two cellulase-positive and five xylanase-positive clones were selected from metagenomic library. Cellulase of clone C1 showed maximal activity at $50^{circ}C$ and pH 6.0, and retained its original activity after 30 min of heat treatment at $60^{circ}C$. Optimum temperature for xylanases of clones X1, X2, X3, and X4 was $50^{circ}C$, and that of clone X5 was $55^{circ}C$. Thermostabilities of xylanases were in the order of X4>X5>X1, X2, and X3. Optimum pH of xylanases of X1, X2, and X3 was 6.0, that of X4 was 5.5, and that of X5 was 5.5-8.0. Xylanasepositive clones could be divided into three groups, X1/X2/X3, X4, and X5, based on influences of temperature and pH on enzyme activity. Sequence analysis of positive subclone of clone C1 showed cellulase, Cel6H, had the highest similarity of 64% to that of Cellulomonas fimi (P07984), suggesting cellulase and xylanase from metagenomic library are novel enzymes.