[Objective] The aim was to study on the electrotransformation of Thermotoga maritima MS8.[Method] Square waves,exponential waves and high voltage shock were used for the electrotransformation of T.maritima MS8,and the...[Objective] The aim was to study on the electrotransformation of Thermotoga maritima MS8.[Method] Square waves,exponential waves and high voltage shock were used for the electrotransformation of T.maritima MS8,and the obtained transformants were detected by PCR.[Result] A single square electric pulse could be applied to cell sample in 0.2 cm ET cuvettes by using a Bio-Rad Gene Pulser set at 150 V,25 ms at room temperature,but the transformation efficiency was very low.[Conclusion] This research may improve the transformation efficiency of T.maritima MS8.展开更多
Gene encoding endo-β-1,4-glucanase(TM1525)is derived from Thermotoga maritima(T.maritima),which has an open reading frame of 825 bp and encodes a 274 amino acid endo-β-1,4-glucanase.This enzyme has the same high tem...Gene encoding endo-β-1,4-glucanase(TM1525)is derived from Thermotoga maritima(T.maritima),which has an open reading frame of 825 bp and encodes a 274 amino acid endo-β-1,4-glucanase.This enzyme has the same high temperature resistance as thermophilic bacteria,which is an ideal property for industrial applications.By molecular biological means,TM1525 was cloned into pHT43 vector and introduced into Bacillus subtilis(B.subtilis)WB800N by electroporation.The results showed that the WB800N expression system was successfully constructed,and extracellular expression of the recombinant gene was achieved.Cellulose hydrolyzed activity of the protein was exhibited.展开更多
将重组海栖热袍菌(Thermotoga maritima)极耐热木聚糖酶B固定于镍螯合环氧载体Eupergit C 250L(固定化XynB)上,水解玉米芯汽爆液连续生产低聚木糖,特别是木二糖。固定化XynB操作稳定性很高,批次水解15次仍保持92%的初始水解活性。90℃...将重组海栖热袍菌(Thermotoga maritima)极耐热木聚糖酶B固定于镍螯合环氧载体Eupergit C 250L(固定化XynB)上,水解玉米芯汽爆液连续生产低聚木糖,特别是木二糖。固定化XynB操作稳定性很高,批次水解15次仍保持92%的初始水解活性。90℃下连续水解168h后保持83.6%的初始水解活性,连续操作半衰期为577.6h(24d)。HPLC分析表明,连续水解液中主要含有49.8%木二糖和22.6%木糖。活性碳层析柱可吸附木二糖,利用乙醇梯度洗脱木二糖的收率为84.7%,纯度为97.2%。因此,固定化XynB可用于水解玉米芯汽爆液高效生产木二糖。展开更多
基金Supported by Sub-Project of"973"(2004CB719600)National Natural Science Foundation Projects(30170511)~~
文摘[Objective] The aim was to study on the electrotransformation of Thermotoga maritima MS8.[Method] Square waves,exponential waves and high voltage shock were used for the electrotransformation of T.maritima MS8,and the obtained transformants were detected by PCR.[Result] A single square electric pulse could be applied to cell sample in 0.2 cm ET cuvettes by using a Bio-Rad Gene Pulser set at 150 V,25 ms at room temperature,but the transformation efficiency was very low.[Conclusion] This research may improve the transformation efficiency of T.maritima MS8.
基金National Natural Science Foundation of China(Regional Fund)(No.51863020)
文摘Gene encoding endo-β-1,4-glucanase(TM1525)is derived from Thermotoga maritima(T.maritima),which has an open reading frame of 825 bp and encodes a 274 amino acid endo-β-1,4-glucanase.This enzyme has the same high temperature resistance as thermophilic bacteria,which is an ideal property for industrial applications.By molecular biological means,TM1525 was cloned into pHT43 vector and introduced into Bacillus subtilis(B.subtilis)WB800N by electroporation.The results showed that the WB800N expression system was successfully constructed,and extracellular expression of the recombinant gene was achieved.Cellulose hydrolyzed activity of the protein was exhibited.
文摘将重组海栖热袍菌(Thermotoga maritima)极耐热木聚糖酶B固定于镍螯合环氧载体Eupergit C 250L(固定化XynB)上,水解玉米芯汽爆液连续生产低聚木糖,特别是木二糖。固定化XynB操作稳定性很高,批次水解15次仍保持92%的初始水解活性。90℃下连续水解168h后保持83.6%的初始水解活性,连续操作半衰期为577.6h(24d)。HPLC分析表明,连续水解液中主要含有49.8%木二糖和22.6%木糖。活性碳层析柱可吸附木二糖,利用乙醇梯度洗脱木二糖的收率为84.7%,纯度为97.2%。因此,固定化XynB可用于水解玉米芯汽爆液高效生产木二糖。
