CAJ | 학술논문

目的对酿酒酵母固有的β-胡萝卜素代谢途径进行扩展，构建能够合成虾青素的酿酒酵母工程菌。 　　方法对不同来源的虾青素合成基因 crtZ（β-胡萝卜素羟化酶）与 crtW （β-胡萝卜素酮化酶）进行随机组合，采用overlap PCR 技术构建串联表达质粒，并将其导入产β-胡萝卜素酿酒酵母工程菌内，发酵产物经 HPLC 和 UPLC-MS鉴定，确定其成分。 　　结果构建出8种 TEF1p-crtZ-ADH1t-PGK1P-crtW-CYC1t组合的串联表达模式，其中基因组合分别为 crtZ1-crtW1， crtZ1-crtW4工程菌 HCCB08571、HCCB08572产生了虾青素；工程菌 HCCB08566、HCCB08568、HCCB08573未检测到产物虾青素，而出现了中间产物玉米黄质和斑蝥黄质。 　　结论成功构建了产生虾青素及其中间体的酿酒酵母工程菌。
목적대양주효모고유적β-호라복소대사도경진행확전，구건능구합성하청소적양주효모공정균。 　　방법대불동래원적하청소합성기인 crtZ（β-호라복소간화매）여 crtW （β-호라복소동화매）진행수궤조합，채용overlap PCR 기술구건천련표체질립，병장기도입산β-호라복소양주효모공정균내，발효산물경 HPLC 화 UPLC-MS감정，학정기성분。 　　결과구건출8충 TEF1p-crtZ-ADH1t-PGK1P-crtW-CYC1t조합적천련표체모식，기중기인조합분별위 crtZ1-crtW1， crtZ1-crtW4공정균 HCCB08571、HCCB08572산생료하청소；공정균 HCCB08566、HCCB08568、HCCB08573미검측도산물하청소，이출현료중간산물옥미황질화반모황질。 　　결론성공구건료산생하청소급기중간체적양주효모공정균。
Objective To construct the engineering strain producing astaxanthin by extending β-carotene metabolism pathway in Saccharomyces cerevisiae. Methods crtZ, crtW genes of different origins were randomly combinated to construct the tandem expression plasmids through overlap PCR techniques, and then transformed into Saccharomyces cerevisiae to obtain the engineered strain with astaxanthin synthesis. Fermented products were identified by HPLC and UPLC-MS to determine their composition. Results 8 patterns of gene multimers TEF1p-crtZ-ADH1t-PGK1P-crtW-CYC1t were constructed, but astaxanthin was expressed by the engineered strains HCCB08571 and HCCB08572 with CrtZ1-CrtW1 and CrtZ1-CrtW4 combinations, respectively. The astaxanthin were not detected in the fermentants of the other engineered strains and the intermediates, zeaxanthin and canthaxanthin were found. Conclusion Saccharomyces cerevisiae strains producing astaxanthin and its intermediates are successfully constructed in this study.