CAJ | 학술논문

将来源于嗜热厌氧乙醇菌(Thermoanaerobacter ethanolicus JW200)的双活性阿拉伯/木糖苷酶(XarB)基因构建到新型热激质粒pHsh上,得到质粒pHsh-xarB。将来源于疏棉状嗜热丝孢菌( Thermomyces lanuginosus DSM 5826)的木聚糖酶A(XynA)基因构建到pHsh-xarB上,得到表达质粒pHsh-xarB-xynA。将重组质粒pHsh-xarB-xynA转入大肠杆菌Escherichia coli JM109进行表达。 SDS-PAGE结果显示,该重组酶的分子量为108000,与理论值相符。Xyn-SDS-PAGE显示该融合酶具备木聚糖酶的活性。基于热激载体 pHsh 的重组表达系统具有诱导表达简便、诱导方式廉价的优点,且重组酶热稳定性好,这对该酶的大规模发酵应用具有重要意义。
장래원우기열염양을순균(Thermoanaerobacter ethanolicus JW200)적쌍활성아랍백/목당감매(XarB)기인구건도신형열격질립pHsh상,득도질립pHsh-xarB。장래원우소면상기열사포균( Thermomyces lanuginosus DSM 5826)적목취당매A(XynA)기인구건도pHsh-xarB상,득도표체질립pHsh-xarB-xynA。장중조질립pHsh-xarB-xynA전입대장간균Escherichia coli JM109진행표체。 SDS-PAGE결과현시,해중조매적분자량위108000,여이론치상부。Xyn-SDS-PAGE현시해융합매구비목취당매적활성。기우열격재체 pHsh 적중조표체계통구유유도표체간편、유도방식렴개적우점,차중조매열은정성호,저대해매적대규모발효응용구유중요의의。
The structure gene from Thermoanaerobacter ethanolicus JW200 encoding XarB was amplified and ligated into pHsh, resulting in pHsh-xarB. The structure gene from Thermomyces lanuginosus encoding xylanase ( XynA) was am-plified and ligated into pHsh-xarB, resulting in pHsh-xarB-xynA. The multi-functional enzyme ( XarB-XynA) was achieved after the expression of pHsh-xarB-xynA in Escherichia coli JM109. SDS-PAGE analysis showed that the molecular mass of the expressed recombinant XarB-XynA was about 108 000, which was exactly the size predicted. The recombinant XarB-XynA exhibited xylanase activity. The expression vector system based on the heat shock vector pHsh owned such advantages as high expression level, cheap induction, and thermo-stability of the recombinant enzyme, which were crucial for the application of large-scale fermentation.