" 第一年的工作為將本實驗室現有之 A. salmonicida MAEF108 或 P. vesicularis MA103 將完成胺基酸定序和基本生化特性測試Agarases，分別用以分解紅藻 (紫菜)、 褐藻 (馬尾藻)、及綠藻 (海菜) 溶液後，分別測定其抗氧化、抗病毒性、抗凝血性、與 微生物之抑菌性。並進一步分析海藻多酚含量、含硫酸酯寡醣含量、及(或) 寡醣之單 糖組成個數與生理活性之相關性，並擬找出 2-3 種含海藻多酚或含硫酸酯之具開發應 用價值海藻寡醣。並擬與本系吳彰哲老師合作探討各類海藻寡醣分解液之抗本土腸病毒 之抑制活性試驗，以做為後續發展應用於保健食品之基礎。 第二年的工作預定為接續前一年度的生理活性測試中的抗致突變性試驗，並綜合生理活 性實驗結果，挑選 2-3 個具有開發潛力及應用價值agarase，依照已獲致之胺基酸定序 結果設計引子，以將具運用潛力之 agarase 基因選殖至如食品級之乳酸菌株中表現，以 及評估該具保健功能乳酸菌之發酵與產生cloned agarase 的表現。並擬與本系吳彰哲老 師合作探討各類海藻寡醣分解液之抗日本腦炎病毒之抑制活性試驗，以做為後續發展應 用於保健食品之基礎。 第三年的工作為將前二年獲得之開發潛力及應用價值的 agarase 基因以 site-directed mutagenesis 等蛋白質工程技術產生新的agarase，並確認其較原來agarase具有更易於利 用和可有效率生產特定海藻寡醣分解液之能力。綜合前二年度以海藻寡醣水解液對病毒 株的抑制實驗結果，組合有效抑制的海藻寡醣水解物，進行腸病毒或日本腦炎的老鼠動 物實驗，以瞭解可能存在的抗病毒反應機制。 " " In the first year of this project, agarases produced by A. salmonicida MAEF108 or P. vesicularis MA103 had been purified and sequenced and tested for their basic biochemical properties. The individual purified agarases will be used to digest red alga (Porphyra dentat), brown alga (Sargussam duplicatum), and green alga (Monostroma nitidum), then their digested lysates were examined by the effects on the antioxidation, antivirus, anticoagulation, and/or the inhibition on microorganisms. Furthermore the valuable components such as algal polyphenolic compounds, sulfated oligosaccharides, and/or the number of monomer of certain oligosaccharides will be measured and their relationships to the biological activities will be investigated. Two to three such algal oligosaccharides will be identified for further application study. The various algal oligosaccharide lysates (AOsL) will be examined with local enteric virus for their inhibition effect in the lab of Assist. Professor C.J. Wu for evaluation their potential use as antivirus health food in the future. In the second year work of this project, the biological evaluation on the antimutagenicity will be conducted. According to the biological activities of AOsLs evaluated, 2-3 corresponding agarases for the production of these biological active AOsLs will be identified. Search for their encoding genes by using the amino acid sequencing data obtained earlier could be used to design a primer for gene cloning and these agarase genes are planed to be expressed in certain food-grade functional lactic acid bacteria (LAB) for future health food development. The performance on fermentation and agarase production of these cloned host LAB will be compared to their parent strains. The various algal oligosaccharide lysates (AOsL) will be examined with Japanese encephalitis virus (JEV) for their inhibition effect in the lab of Assist. Professor C.J. Wu for evaluation their potential use as antivirus health food in the future. During the third year, the agarase gene that have been identified to be good potential for use in health food products will be the modified by protein engineering techniques such as site-directed mutagenesis to make new agarase. These new agarases will be confirmed for their capabilities on easier applications for AOsLs production and producing specified AOsLs for health food. After considering the inhibition results of previous results on the enteric virus and JEV of specific AOsLs, the right combination of AOsLs will be tested via rat animal experiments. And the inhibition mechanism of AOsL to the virus may be solved. "
health biological characteristics
Japanese encephalitis virus (JEV)