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| 嗜热糖化酶基因在重组黑曲霉工程菌中的表达及酶学性质初步研究 | |
| 引用本文： | 王凤寰,孟青青,封棣,杨建国,汪兵.嗜热糖化酶基因在重组黑曲霉工程菌中的表达及酶学性质初步研究[J].生物技术通报,2012(7):119-125. |
| 作者姓名： | 王凤寰 孟青青 封棣 杨建国 汪兵 |
| 作者单位： | 1. 北京工商大学食品学院食品添加剂与配料北京高校工程研究中心北京市食品风味化学重点实验室,北京,100048 2. 北京化工大学生命科学与技术学院,北京,100029 3. 北京中天诺亚酶制剂有限公司,北京,102629 |
| 摘 要： | 为了提高糖化酶的耐热性能,降低淀粉糖化发酵工艺的生产成本,构建了同源整合载体pEasy-glaAdir以及pEasyssg,将黑曲霉(Aspergillus niger)的糖化酶基因(glaA)灭活,并将硫磺矿硫化叶菌(Sulfolobus solfataricus)的嗜热糖化酶基因(ssg)插入到黑曲霉基因组中,筛选得到表达嗜热糖化酶的重组黑曲霉工程菌(A.nigerWW1)。重组菌的发酵结果显示,嗜热糖化酶在黑曲霉中得到了分泌表达,发酵液酶活达到3 030 U/mL。重组嗜热糖化酶的最适反应温度为90℃,最适pH为6.0,该酶具有较高的热稳定性,在80℃时的半衰期在60 min以上,具有良好的工业应用前景。 |
| 关 键 词： | 黑曲霉 硫磺矿硫化叶菌 嗜热糖化酶 同源重组 热稳定性 |
Expression of Thermostable Glucoamylase Gene in Aspergillus niger and Its Characterization |
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| Wang Fenghuan , Meng Qingqing , Feng Di , Yang Jianguo , Wang Bing.Expression of Thermostable Glucoamylase Gene in Aspergillus niger and Its Characterization[J].Biotechnology Bulletin,2012(7):119-125. | |
| Authors: | Wang Fenghuan Meng Qingqing Feng Di Yang Jianguo Wang Bing |
| Institution: | 1Beijing Higher Institution Engineering Research Center of Food Additives and Ingredients,Beijing Key Laboratory of Food Flavor Chemistry,School of Food and Chemical Engineering,Beijing Technology and Business University(BTBU),Beijing 100048;2College of Life Science and Technology,Beijing University of Chemical Technology,Beijing 100029;3Beijing Zhongtian Nuoya Enzyme Co.,Ltd.,Beijing 102629) |
| Abstract: | It was to improve the glucoamylase thermostability and reduce the cost of industrial fermentation,integrative vectors pEasy-glaAdir and pEasy-ssg were constructed and transformed into Aspergillus niger CICC 2204.The native glaA gene was interrupted and the Sulfolobus solfataricus glaA gene(ssg) encoding the thermostable glucoamylase was integrated into the chromosome of A.niger,which yielded a thermostable glucoamylase producer.The positive transformant showed great secretive expression of SSG and the enzyme activity reached 3 030 U/mL.The recombinant thermostable glucoamylase exhibited optimum catalytic activity at 90℃ and pH6.0,respectively,and remained 50% of its original activity after 60 min at 80℃.It was more thermostable than that of native GLA. |
| Keywords: | Aspergillus niger Sulfolobus solfataricus Glucoamylase Homologus recombination Thermostability |
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