Kim Suyeon, Kim Yeon-Ok, Lee Yongjik, Choi Inseong, Joshi Chandrashekhar P, Lee Kyehan, Bae Hyeun-Jong
Bio-Energy Research Institute, Chonnam National University, Gwangju, Korea.
Biosci Biotechnol Biochem. 2012;76(6):1140-5. doi: 10.1271/bbb.110981. Epub 2012 Jun 7.
Plants are attractive expression systems for large-scale, low-cost production of high-value proteins. The xylanase 2 gene (Xyn2), encoding an endo-β-1,4-xylanase from Trichoderma reesei, was cloned and expressed in Escherichia coli and the poplar (Populus spp.). The optimal temperature and pH of the recombinant xylanase were 50 °C and 5.0 respectively when expressed in E. coli. The purpose of this study was to produce recombinant xylanase in poplar. The Xyn2 gene was transferred into poplars by Agrobacterium-mediated transformation. The transgenic status and transgene expression of the transformed poplar were confirmed by polymerase chain reaction (PCR) genotyping and reverse transcription (RT)-PCR analysis. The poplar-expressed xylanase was biologically active, with an expression level of up to 14.4% of total leaf soluble protein. In the leaves, the average xylanase content was 1.016 mg per g of leaf fresh weight in the transgenic poplar. We found that the poplar might make possible the large-scale production of commercially important recombinant proteins.
植物是用于大规模、低成本生产高价值蛋白质的理想表达系统。编码里氏木霉内切-β-1,4-木聚糖酶的木聚糖酶2基因（Xyn2）被克隆并在大肠杆菌和杨树（Populus spp.）中表达。在大肠杆菌中表达时，重组木聚糖酶的最适温度和pH分别为50℃和5.0。本研究的目的是在杨树中生产重组木聚糖酶。通过农杆菌介导的转化将Xyn2基因导入杨树。通过聚合酶链反应（PCR）基因分型和逆转录（RT）-PCR分析确认了转化杨树的转基因状态和转基因表达。杨树表达的木聚糖酶具有生物活性，表达水平高达总叶可溶性蛋白的14.4%。在叶片中，转基因杨树中木聚糖酶的平均含量为每克叶片鲜重1.016毫克。我们发现杨树可能使大规模生产具有商业重要性的重组蛋白成为可能。
