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KMID : 0613820060160020360
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Journal of Life Science
2006 Volume.16 No. 2 p.360 ~ p.364
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Screening of Fibrinolytic Enzymes from Soil Metagenome Library
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Lee Sun-Yi
Kim Bo-Hye
Kang Ju-Hyung
Cho Hyo-Jin
Kong Eun-Hee
Moon Sang-Wook
Kim Young-Jin
Ahn Soon-Cheol
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Abstract
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Fibrin clots of blood vessels are one of the serious factor caused cardiovascular disease. The development of a antithrombotic and thrombolysis solvent is necessary to prevent and treat these diseases. It has been reported that a strong fibrin-specific fibrinolytic enzyme was produced from a Korean fermented soybean paste similar to Japanese miso. We have been screened the known or novel fibrinolytic enzymes by activity-based and sequence-based screening from soil DNA metagenome library containing all kinds of environmental genomic DNA. The activity-based screening was determined the protease activity on 0.5% skim milk. For sequence-based screening, we designed a set of primer expanding gene sequence of fibrinolytic enzyme, performed PCR and selected clones showing the expected size of amplicons from metagenome library. Transformation of the gene encoding fibrinolytic enzyme was carried out with commercial vectors and their transformants were selected. Finally, we found 15 positive clones from metagenome library. Then each of sequences were analyzed and identified as similar or known the clones of nattokinase. We are going to perform full sequence of each clones, ligate with expression vector, transform into competent cells and then determine activity of expressed enzymes.
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KEYWORD
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Fibrin, fibrinolytic, soil metagenome, screening, protease
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