SCIENCE

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Browsing SCIENCE by Subject "(AFRICAN YAM BEANS),"

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    ISOLATION, PARTIAL PURIFICATION, CHARACTERIZATION AND ANTIFUNGAL POTENTIAL OF CHITINASE FROM SEEDS OF SPHENOSTYLIS STENOCARPA (AFRICAN YAM BEANS)
    (2017-03) OBIAKOR, Chimee Ethel
    The utilization of chemical fungicide has attracted increased scrutiny since they cause environmental contamination and induce pathogen resistance. These limitations calls for harmless alternative control strategy to prevent plant diseases for sustainable agriculture. This work was designed to partially purify,characterize and tested for the antifungal potentials of chitinase from seeds of African Yam Beans. Chitinase (EC.3.2.14) was isolated from Stephynostylis stenocarpa, by precipitation using 80% Ammonium Sulphate Saturation and Gel filtration on Sephadex G-75 column to 8.04 fold with a yield of 46.35% and a final specific activity of 4.45μmol/min. SDS-PAGE of the enzyme showed a molecular weight of 32KDa. The enzyme had an optimal temperature and pH of 55oC and 5.0 respectively. It had temperature and pH stability within the range of 30-60oC and 4-7 respectively. Initial velocity studies for the determination of kinetic constants with colloidal chitin as substrate revealed a KM and Vmax of 1.2mMand 12.29μmol/min. The Chitinase activity was inhibited by Pb2+, Hg2+, K+ and other reagents like EDTA and glucose but Na+ and Zn2+ showed little or no effect on the enzyme activity. The antifungal potentials of this Chitinase was tested against Fusarium oxysponium, Fusarium solani, and Alterneria alternate. It had no effect on the Fusarium species but inhibited the growth of Alterneria alternate as seen from clear Zones of inhibition (14.00mm-17.00mm) for the crude extract and (9.00mm-11.00mm) for the purified using agar diffusion method. This indicates that the enzyme can be used to combat such diseases triggered by the organism e.g. leaf blight, hence servingas a tool for crop protection in the agricultural sector