Purification and Characterization of Two Thermostable Glucoamylases Produced from Aspergillus niger B-30

doi:10.1007/s40242-013-3074-y

高等学校化学研究 ›› 2013, Vol. 29 ›› Issue (5): 917-923.doi: 10.1007/s40242-013-3074-y

Purification and Characterization of Two Thermostable Glucoamylases Produced from Aspergillus niger B-30

LIU Yang^1,2, LI Quan-shun¹, ZHU Hong-liang³, MENG Zhao-li^1,2, XIANG Hong-yu^1,2, XIE Qiu-hong^1,2

1. Key Laboratory for Molecular Enzymology and Engineering, Ministry of Education, Changchun 130012, P. R. China;
2. National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, P. R. China;
3. Zibo Guoao Biotechnology Co., Ltd., Zibo 255400, P. R. China

收稿日期:2013-02-27 修回日期:2013-06-13 出版日期:2013-10-01 发布日期:2013-09-17
通讯作者: XIANG Hong-yu E-mail:hyxiang@jlu.edu.cn
基金资助:
Supported by the National Natural Science Foundation of China(No.81072564) and the Project of the Jilin Province Science and Technology Institute of China(No.20090945).

Purification and Characterization of Two Thermostable Glucoamylases Produced from Aspergillus niger B-30

LIU Yang^1,2, LI Quan-shun¹, ZHU Hong-liang³, MENG Zhao-li^1,2, XIANG Hong-yu^1,2, XIE Qiu-hong^1,2

1. Key Laboratory for Molecular Enzymology and Engineering, Ministry of Education, Changchun 130012, P. R. China;
2. National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, P. R. China;
3. Zibo Guoao Biotechnology Co., Ltd., Zibo 255400, P. R. China

Received:2013-02-27 Revised:2013-06-13 Online:2013-10-01 Published:2013-09-17
Contact: XIANG Hong-yu E-mail:hyxiang@jlu.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China(No.81072564) and the Project of the Jilin Province Science and Technology Institute of China(No.20090945).

摘要/Abstract

摘要：

Two thermostable glucoamylases were produced from Aspergillus niger B-30 by submerged fermentation. The two glucoamylases GAM-1 and GAM-2 were purified by ammonium sulfate precipitation, diethylaminoethylcellulose fast flow(DEAE FF) and Superdex G-75 gel filtration columns. The molecular weights of GAM-1 and GAM-2 were determined as 9.72×10⁴ and 7.83×10⁴ by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), while the molecular weights of GAM-1 and GAM-2 were determined to be 8.05×10⁴ and 7.04×10⁴ by matrix assisted laser desorption ionizationtime-of-flight(MALDI-TOF) mass spectrometry, respectively. Both the enzymes were glycosylated, with 10.4% and 11.4% carbohydrate content, respectively. The optimal pH and temperature were 4.0―4.6 and 70 ℃ for both. The two glucoamylases were maintained 100% relative activity after incubation at 60 ℃ for 120 min. After the hydrolysis of starch for 120 min, glucose was the only product, confirming that the two enzymes were of high efficiency towards starch. The GAM-2 exhibited higher catalytic activity towards oligosaccharides such as maltose than GAM-1, and the kinetic analysis shows that the affinity of GAM-2 to starch was lower than that of GAM-1. The high thermostability and effectiveness make the two glucoamylases potentially attractive for biotechnological application.

关键词: Glucoamylase, Thermostability, Kinetic analysis, Glycosylation

Abstract:

Key words: Glucoamylase, Thermostability, Kinetic analysis, Glycosylation

LIU Yang, LI Quan-shun, ZHU Hong-liang, MENG Zhao-li, XIANG Hong-yu, XIE Qiu-hong. Purification and Characterization of Two Thermostable Glucoamylases Produced from Aspergillus niger B-30[J]. 高等学校化学研究, 2013, 29(5): 917-923.

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Purification and Characterization of Two Thermostable Glucoamylases Produced from Aspergillus niger B-30

Purification and Characterization of Two Thermostable Glucoamylases Produced from Aspergillus niger B-30

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