CAJ | 학술논문

木糖醇脱氢酶（ xylitol dehydrogenase，XDH）可以氧化木糖醇生成木酮糖，处于木糖代谢的节点位置。利用PCR方法克隆得到了休哈塔假丝酵母（ Candida shehatae）20335的木糖醇脱氢酶基因、质粒pKT0150的ADH1终止子序列和G418抗性基因（ KanR），以及酿酒酵母（ Saccharomyces cerevisiae）W5特定的2.2 kb的rDNA片段。以酿酒酵母整合载体p406ADH1为骨架，利用基因工程手段构建一个多拷贝整合表达载体pLX-AGRX。将重组载体pLX-AGRX线性化转入到酿酒酵母W5后，通过高浓度G418筛选和PCR双重鉴定，证实重组载体pLX-AGRX已整合到酿酒酵母W5基因组上，测定木糖醇脱氢酶酶活可达65.9574 U/mg。
목당순탈경매（ xylitol dehydrogenase，XDH）가이양화목당순생성목동당，처우목당대사적절점위치。이용PCR방법극륭득도료휴합탑가사효모（ Candida shehatae）20335적목당순탈경매기인、질립pKT0150적ADH1종지자서렬화G418항성기인（ KanR），이급양주효모（ Saccharomyces cerevisiae）W5특정적2.2 kb적rDNA편단。이양주효모정합재체p406ADH1위골가，이용기인공정수단구건일개다고패정합표체재체pLX-AGRX。장중조재체pLX-AGRX선성화전입도양주효모W5후，통과고농도G418사선화PCR쌍중감정，증실중조재체pLX-AGRX이정합도양주효모W5기인조상，측정목당순탈경매매활가체65.9574 U/mg。
Beinginanodepositioninxylosemetabolism,xylitoldehydrogenase(XDH)isabletooxidatexylitolinto xylulose. In this experiment the XDH gene xyl2,ADH1 terminal subsequence and resistant gene G418( KanR)of palsmid pKT0150 from the genome of Candida shehatae 20335 using PCR cloning,as well as specific 2. 2 kb rDNA fragment of S. serevisiae W5 were obtained. The multicopying integrated expression vector pLX-AGRX was constructed using integrated vector p406ADH1 of S. serevisiae as a framework by means of genetic engineering. The recombinant vector pLX-AGRX was linearized and transferred into S. cerevisiae W5,then screened through high concentration G418 and PCR for double characterization,and proved that the recombinant vector pLX-AGRX was integrated into ge-nome of S. serevisiae W5,it was tested that the activity of XDH was as high as 65. 957 4 U/mg.