초록 ▼

Ⅳ. 연구개발결과
1. 신규 흑염소 반추위 미생물 메타게놈 해독
pyrosequencing 1.9Gb와 illumina NGS short read sequencing 300Gb 데이터에 대한 de novo assembly 수행 및 유전자 탐색
2. 섬유소분해 활성 흑염소 반추위 메타게놈 fosmid 클론 염기서열 분석
산업재산권 출원된 34개 유전자를 포함하는 fosmid 클론 총 19개의 전장 염기서열 해독을 수행하여 유전체 구조 특성 분석을 수행한 결과, 총 39개의 섬유소분해효소 유전자가 예측되었으며,

Ⅳ. 연구개발결과
1. 신규 흑염소 반추위 미생물 메타게놈 해독
pyrosequencing 1.9Gb와 illumina NGS short read sequencing 300Gb 데이터에 대한 de novo assembly 수행 및 유전자 탐색
2. 섬유소분해 활성 흑염소 반추위 메타게놈 fosmid 클론 염기서열 분석
산업재산권 출원된 34개 유전자를 포함하는 fosmid 클론 총 19개의 전장 염기서열 해독을 수행하여 유전체 구조 특성 분석을 수행한 결과, 총 39개의 섬유소분해효소 유전자가 예측되었으며, 앞서 29개의 섬유소분해효소 유전자 탐색 결과와 비교하여 10개가 많은 것으로 확인되었다.

Abstract ▼

1st detailed assignment : Competitiveness of domestic animal farmers has been declined according to increase of fodder import after FTA with many countries of powerful livestock industry. To improve the feed efficiency and supply cellulases with low cost, novel active cellulase genes have been inves

1st detailed assignment : Competitiveness of domestic animal farmers has been declined according to increase of fodder import after FTA with many countries of powerful livestock industry. To improve the feed efficiency and supply cellulases with low cost, novel active cellulase genes have been investigated. Korean black goat has been evolutionally well adapted for roughages rather than other ruminants. Therefore, the goat rumen microbiome might be well developed to digest cellulosic materials. We performed the construction of fosmid libraries and massive sequencing of rumen microbial metagenomes from Korean black goats. A total of 34 novel cellulase genes were found and clones into expression vectors. When the novel cellulases were induced, most of them were expressed with insoluble form. We investigated the refolding condition of the novel cellulases. Generally, CMCase activities were shown form 2,000 to 4,000 units/ml and xylanase activities from 5,000 to over 20,000 units/ml. Also, up to 4,000 units/ml of xylanase activities were measured in the supernatant media of several cellulases. Optimal activities of most of the novel cellulases were shown at pH 6 with 50℃. Over 60% of activities of them were kept at from 30℃ to 60℃ in the condition of pH 6. In the result of digestion efficiency in vitro experiment by addition of novel cellulases, significant increase of digestion efficiency was not shown, but 20% of VFA generation was increased after incubation for 48 hrs. In conclusion, external secretion system should be developed for industrial application of these novel cellulases.
2nd detailed assignment : The objective of this study was to determine the effect of dietary addition of a fibrolytic enzyme derived from rumen microbial metagenome of black goat. The use of cellulolytic enzyme increases the degradation of fiber in the feed. It is mainly used to increase the productivity of livestock. Rode et al. (1999) showed that enzymes improved digestibility of DM and fiber from dairy diets. Feng et al. (1996) found that fibrolytic enzymes improved DM digestibility in both in vitro and in situ studies. However, other studies did not (Beauchemin et al., 2000; Kung et al., 2002; Sutton et al., 2003). This studies were conducted to investigate the effects of fibrolytic enzyme derived from rumen microbial metagenome of black goat. Italian ryegrass, rice straw were used as substrates. In an additional incubating substrates with the enzymes but without rumen fluid was studied. we measured in vitro dry matter disappearance, volatile fatty acid, pH and gas production. The measured results showed no difference compared without cellulolytic enzyme.