CAJ | 학술논문

利用低能N＋注入介导秦艽基因组DN A转化多形汉逊酵母技术，结合颜色反应、薄层层析和高效液相色谱等方法筛选重组菌株，获得1株遗传稳定的能生物合成龙胆苦苷的重组酵母菌，被命名为DL-49．经液体培养90 h后，利用 HPLC分析其发酵液中的龙胆苦苷，其产量为2．22mg／L ．将重组菌株传代培养8代后，分析显示重组酵母菌生物合成龙胆苦苷的产量基本稳定．试验表明，采用低能离子注入介导秦艽基因组转化酵母技术，可以在龙胆苦苷生物合成相关基因信息未知的情况下，直接获得能生物合成龙胆苦苷的酵母重组菌．
이용저능N＋주입개도진구기인조DN A전화다형한손효모기술，결합안색반응、박층층석화고효액상색보등방법사선중조균주，획득1주유전은정적능생물합성룡담고감적중조효모균，피명명위DL-49．경액체배양90 h후，이용 HPLC분석기발효액중적룡담고감，기산량위2．22mg／L ．장중조균주전대배양8대후，분석현시중조효모균생물합성룡담고감적산량기본은정．시험표명，채용저능리자주입개도진구기인조전화효모기술，가이재룡담고감생물합성상관기인신식미지적정황하，직접획득능생물합성룡담고감적효모중조균．
To obtain an alternative source for the production of Gentiopicroside ,a very simple method for achieving one-step whole-pathway assembly of Gentiopicroside biosynthesis in yeast is presented .Genomic DNA segments of medicinal plant Gentiana macrophylla (G . macrophylla) were randomly transferred into Hansenula polymorpha (H .polymorpha) by 25 KeV nitrogen ions (N+ ) at a dose of 2 .0 × 1016 ions/cm2 under the vacuum pressure of 1 × 10-3 Pa .One potential stable recombinant yeast strain capable of producing Gentiopicro-side was obtained using a combination screening of Fehling′s test and TLC ,and designated as DL-49 .The potential Gentiopicroside-producing strain DL-49 was further analyzed by HPLC ,and the result showed that the sample from strain DL-49 possessed of a retention time and ion peaks consistent with those of the Gentiopicroside standard .The corresponding Gentiopicroside yield was 2 .22 mg/L as determined by HPLC .Additionally ,the recombinant strain was stable for at least 8 generations .One Gentiopicroside-producing recombinant strain was obtained by low energy N+ ion implantation .This would have a valuable applica-tion for low-cost production of natural compounds .