보고서 정보
주관연구기관 |
한국생명공학연구원 Korea Research Institute of Bioscience and Biotechnology |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2011-12 |
과제시작연도 |
2010 |
주관부처 |
농림축산식품부 Ministry of Agriculture, Food and Rural Affairs(MAFRA) |
등록번호 |
TRKO201400026433 |
과제고유번호 |
1545002477 |
사업명 |
농림기술개발 |
DB 구축일자 |
2014-11-10
|
DOI |
https://doi.org/10.23000/TRKO201400026433 |
초록
▼
○ 연구결과
<논문발표 2건>
- Cloning and characterization ofamodularGH5 β-1,4-mannanasewith high specificactivity from the fibrolytic bacterium Cellulosimicrobium sp.strain HY-13.Bioresource Technology 102,9185-9192.2011.(IF :4.365)
- Novelmodularendo-β-1,4-xylanasewith transglycosylation activity fr
○ 연구결과
<논문발표 2건>
- Cloning and characterization ofamodularGH5 β-1,4-mannanasewith high specificactivity from the fibrolytic bacterium Cellulosimicrobium sp.strain HY-13.Bioresource Technology 102,9185-9192.2011.(IF :4.365)
- Novelmodularendo-β-1,4-xylanasewith transglycosylation activity from Cellulosimicrobium sp.strain HY-13 that Is homologous to inverting GH family 6 enzymes.Bioresource Technology(inpress)2011.(IF :4.365)
<특허출원 1건,PCT 진입 1건>
- 출원번호 10-2011-0146600:셀룰로시마이크로비움 속 HY-13균주로부터 생산되는 신규한 활성 도메인을 갖는 자일라나제
-PCT 진입 1건 :셀룰로시마이크로비움 속 HY-13 균주로부터 생산된 신규한 만난아제(국내 특허 출원 10-2011-0070462)
Abstract
▼
Ⅳ. Results
◯ A xylanolytic microorganism, strain HY-22 was isolated from a compost sample and phylogenetically identified as a Gram-positive bacterium belonging to the genus Bacillus. When cultivated on a liquid medium that contained 2.0% wheat bran (80 mesh passed), 0.9% yeast extract, 0.9% pept
Ⅳ. Results
◯ A xylanolytic microorganism, strain HY-22 was isolated from a compost sample and phylogenetically identified as a Gram-positive bacterium belonging to the genus Bacillus. When cultivated on a liquid medium that contained 2.0% wheat bran (80 mesh passed), 0.9% yeast extract, 0.9% peptone, 0.2% KNO3, 0.01% MgSO4, and 0.1% olive oil, Bacillus sp. strain HY-22 produced xylanase (XylC) a ta yield of approximately 228 IU/ml.
◯ XylC was purified to electrophoretic homogeneity by ammonium sulfate precipitation and two consecutive rounds of column chromatography using DEAE Sephadex A-50 and Superdex G-75 resins. The relative molecular mass of the purified enzyme was approximately 24 kDa, as determined by SDS-PAGE.
◯ XylC exhibited the highest catalytic activity toward birchwood xylan when it was reacted with the substrate at pH 6.5 and 50oC. Of the evaluated chemical compounds, Trp-directed modifiers, Hg2+ (1 mM) and N-Bromosuccinimide (5 mM) were highly toxic to XylC.
◯ XylC displayed a high catalytic activity towards oat spelt xylan, birchwood xylan, and beechwood xylan, but avicel and locust bean gum were not hydrolyzed.
◯ The gene (1,272-bp) encoding a β-1,4-mannanase from a gut bacterium of Eisenia fetida, Cellulosimicrobium sp. strain HY-13 was cloned and expressed in Escherichia coli.
◯ The recombinant β-1,4-mannanase (rManH) was approximately 44.0 kDa and contained a catalytic GH5 domain that was 65% identical to that of the Micromonospora sp. β-1,4-mannosidase.
◯ The enzyme exhibited the highest catalytic activity toward mannans at 50oC and pH 6.0. rManH displayed a high specific activity of 14,711 IU mg-1 and 8,498 IU mg-1 towards ivory nut mannan and locust bean gum, respectively; however it could not degrade structurally unrelated polysaccharides, mannobiose, or p-nitrophenyl sugar derivatives.
◯ rManH was strongly bound to ivory nut mannan, Avicel, chitosan, and chitin but did not attach to curdlan, insoluble oat spelt xylan, lignin, or poly(3-hydroxybutyrate).
◯ Considering its physico-chemical and catalytic properties, ManH, which displayed a maximum activity at pH 6.0, can be exploited as a liquid fermentation product [e.g. Hemicell® (ChemGen Corp., USA)] since it is genetically recognized as a safe (GRAS) organism for herbivorous animals, such as calves, in the feed industry because fibrolytic enzymes in the rumen of herbivorous animals display the highest hydrolytic activity between pH 6.0 and 6.5.
◯ Mixed enzyme complex, Joazyme improved the feed conversion and egg production in broiler and layer. In addition, it reduced the amount of ammonia nitrogen in the cecum.
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