Zhang Hong-Lian, Yao Bin, Wang Ya-Ru, Yuan Tie-Zheng, Zhang Wang-Zhao, Wu Ning-Feng, Fan Yun-Liu
Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Sheng Wu Gong Cheng Xue Bao. 2003 Jan;19(1):41-5.
The gene xynA encoding xylanase was cloned from Streptomyces olivaceoviridis A1. The xynA with and without origin signal peptide sequence were fused behind pel B signal peptide in the plasmid pET-22b(+) respectively, then transfered into the host E. coli. The xylanase expressed in E. coli had normal bioactivity. Further, the xynA without origin signal peptide sequence was cloned into the plasmid pPIC9 under the control of AOX1 promoter and introduced into the host Pichia pastoris by electroporation. The results of SDS-PAGE and activity assay of the xylanase expressed by recombinant P. pastoris showed that the xynA had been overexpressed and secreted, and the xylanase expressed had normal bioactivity. The expression level of xylanase in recombinant P. pastoris exceeded 0.2mg/mL in shake culture.
从橄榄绿链霉菌A1中克隆了编码木聚糖酶的基因xynA。将带有和不带有原始信号肽序列的xynA分别融合到质粒pET-22b(+)中的pel B信号肽之后，然后转入宿主大肠杆菌。在大肠杆菌中表达的木聚糖酶具有正常的生物活性。此外，将不带有原始信号肽序列的xynA克隆到受AOX1启动子控制的质粒pPIC９中，并通过电穿孔导入宿主毕赤酵母。重组毕赤酵母表达的木聚糖酶的SDS-PAGE和活性测定结果表明，xynA已被过量表达并分泌，且表达的木聚糖酶具有正常的生物活性。在摇瓶培养中，重组毕赤酵母中木聚糖酶的表达水平超过0.2mg/mL。
