CAJ | 학술논문

目的：研究真核表达的textilinin-1蛋白纯化品对纤溶酶抑制作用。方法根据textilinin-1的天然氨基酸序列，按照毕氏酵母偏好密码子进行优化，合成 textilinin-1基因，重组到pPICZα载体上，并转化至 Pichia p.X-33菌种中，实现高效分泌表达，并进行重组 textilinin-1活力单位的定义及小鼠断尾试验。结果通过高密度发酵及两步柱层析纯化，最终从10 L发酵液中得到6 g纯度达97.0％以上的重组textilinin-1。活性研究表明，重组 textilinin-1对纤溶酶的活性具有抑制作用，并对 tPA 所致的小鼠出血倾向有一定的抑制作用。结论重组 textilinin-1可抑制纤溶酶活性，并对纤溶性出血小鼠模型有止血作用，具有开发为止血药的潜力。
목적：연구진핵표체적textilinin-1단백순화품대섬용매억제작용。방법근거textilinin-1적천연안기산서렬，안조필씨효모편호밀마자진행우화，합성 textilinin-1기인，중조도pPICZα재체상，병전화지 Pichia p.X-33균충중，실현고효분비표체，병진행중조 textilinin-1활력단위적정의급소서단미시험。결과통과고밀도발효급량보주층석순화，최종종10 L발효액중득도6 g순도체97.0％이상적중조textilinin-1。활성연구표명，중조 textilinin-1대섬용매적활성구유억제작용，병대 tPA 소치적소서출혈경향유일정적억제작용。결론중조 textilinin-1가억제섬용매활성，병대섬용성출혈소서모형유지혈작용，구유개발위지혈약적잠력。
Objective To investigate the inhibition of textilinin-1 ,purified protein of eukaryotic expression ,to the plasmin . Methods On basis of the nature amino acid sequence of textilinin-1 and preference codon in Pichia pas-toris ,the synthetic textilinin-1 gene ,restructuring to the pPICZαcarrier ,and into Pichia p .X-33 strains ,was highly secretory expressed .The definition of recombinant textilinin-1 active unit and tail cutting test in mice were also studied . Results 6 g purity of more than 97.0% of recombinant textilinin-1 was received from 10 L fermentation broth by high density fermentation and two-step column chromatography purification .Activity research indicated that recombinant textilinin-1 can inhibit plasmin ,and can also inhibit bleeding caused by tPA in mice . Conclusion Recombinant textilinin-1 inhibits plasmin ,and has hemostatic effect on fibrinolytic hemorrhages mouse models , so it has potential to be used to prevent or reduce bleeding in surgical .