Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
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Comparison of the Influence of Carbon Substrates on the Fibrolytic Activities of Neocallimastix sp. NLRI-3

탄소원의 종류가 반추위 혐기 곰팡이 Neocallimastix sp. NLRI-3의 섬유소 분해효소 활력에 미치는 영향 비교

  • Published : 2006.06.30
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Abstract

The purpose of this study was to investigate the fungal growth and enzyme production under different carbohydrate substrate conditions. The anaerobic fungus Neocallimastix sp. NLRI-3 isolated from the rumen of Korean native goat was incubated with different carbohydrate media containing 0.2% of glucose, starch, rice straw, filter paper, carboxymethyl cellulose(CMC), Sigmacell cellulose, xylan or xylose, respectively. The culture head gas production was the highest in the culture of filter paper medium, and the lowest in the culture of CMC medium at 96h incubation (P<0.05). The fungal zoospore production reached peak at 72h incubation, and its number was the highest in rice straw medium among the treatments (P<0.05). At 96h incubation, carboxymethyl cellulase(CMCase) activity was the highest in the culture of filter paper medium and the lowest in the culture of starch medium (P<0.05). While xylanase activity was the highest in the culture of rice straw medium and the lowest in the culture of xylose medium(P<0.05) at 72h incubation. There were no differences in culture supernatant protein expression among the treatments. However, the patterns of enzyme expression were different among the treatments with zymogram analysis. Six CMCases and 4 xylanase were detected from the results of zymogram analysis. Therefore the present study indicating that the fungal enzyme expression could be stimulated with insoluble substrates in the culture medium.

한국 재래산양에서 분리된 Neocallimastix sp NLRI-3을 이용하여 기질의 종류에 따른 미생물의 성장 특성 및 효소단백질의 발현양상을 비교하고자 본 연구를 수행하였다. 혐기배지에 glucose, starch, 볏짚, filter paper, CMC, sigmacell, xylan 및 xylose를 0.2% 수준으로 첨가하여 배양하였다. 배양결과 가스 발생량은 96시간대에서 filter paper 배지에서 가장 높았고 CMC배지에서 가장 낮게 조사되었다(P<0.05). 곰팡이 포자 발생량은 72시간대에서 살펴보면 처리구들 중 볏짚 배지에서 가장 높게 나타났다 (P<0.05). 곰팡이가 분비하는 enzyme activity는 보통 72시간대 이후부터 급격히 증가함을 보이는데 CMCase의 경우에 filter paper disc, sigmacell, xylose에서 활성이 높게 나타났다. xylanase 역가는 starch, rice straw, sigmacell에서 높게 나타났다. Zymogram을 이용하여 각 효소단백질의 발현을 분석한 결과 효소단백질의 발현양상은 전 처리구에 걸쳐 유사하였으나 발현정도는 기질의 종류에 따라 차이가 있음을 알 수 있었다. 또한, zymogram 결과에 의해 6개의 CMCase와 4개의 xylanase가 검출 되었다. 본 연구의 결과로 혐기성 곰팡이의 효소단백질은 배양액에 존재하는 기질의 종류에 의해 발현정도가 영향을 받음을 알 수 있었다. 이러한 결과는 산업적으로 이용 가능한 혐기곰팡이 유래 효소제의 개발에 필요한 배양조건의 결정에 이용될 수 있을 것으로 사료된다.

Keywords

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