Overexpression and Biochemical Characterization of an Endo-alpha-1,4-polygalacturonase from Aspergillus nidulans in Pichia pastoris
文献类型:期刊论文
作者 | Xu, Hua4; Zhang, Pengfei3; Zhang, Yuchen2; Liu, Zebin1; Zhang, Xuebing2; Li, Zhimin4; Li, Jian-Jun2; Du, Yuguang2 |
刊名 | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES |
出版日期 | 2020-03-01 |
卷号 | 21期号:6页码:12 |
关键词 | endo-alpha-1,4-polygalacturonase Aspergillus nidulans pH and temperature-rate profile pH and thermal stability identification of hydrolysis products structure modelling |
DOI | 10.3390/ijms21062100 |
英文摘要 | Pectinases have many applications in the industry of food, paper, and textiles, therefore finding novel polygalacturonases is required. Multiple sequence alignment and phylogenetic analysis of AnEPG (an endo-alpha-1,4-polygalacturonase from Aspergillus nidulans) and other GH 28 endo-polygalacturonases suggested that AnEPG is different from others. AnEPG overexpressed in Pichia pastoris was characterized. AnEPG showed the highest activity at pH 4.0, and exhibited moderate activity over a narrow pH range (pH 2.0-5.0) and superior stability in a wide pH range (pH 2.0-12.0). It displayed the highest activity at 60 degrees C, and retained >42.2% of maximum activity between 20 and 80 degrees C. It was stable below 40 degrees C and lost activity very quickly above 50 degrees C. Its apparent kinetic parameters against PGA (polygalacturonic acid) were determined, with the K-m and k(cat) values of 8.3 mg/mL and 5640 mu mol/min/mg, respectively. Ba2+ and Ni2+ enhanced activity by 12.2% and 9.4%, respectively, while Ca2+, Cu2+, and Mn2+ inhibited activity by 14.8%, 12.8%, and 10.2% separately. Analysis of hydrolysis products by AnEPG proved that AnEPG belongs to an endo-polygalacturonase. Modelled structure of AnEPG by I-TASSER showed structural characteristics of endo-polygalacturonases. This pectinase has great potential to be used in food industry and as feed additives. |
WOS关键词 | ACIDIC ENDO-POLYGALACTURONASE ; POTENTIAL APPLICATION ; CRYSTAL-STRUCTURE ; EXPRESSION ; NIGER ; ENDOPOLYGALACTURONASE ; IDENTIFICATION ; PURIFICATION ; ENZYMES ; PECTIN |
资助项目 | National Key Research and Development Program of China[2017YFD0200902] ; National Natural Science Foundation of China[31370799] |
WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry |
语种 | 英语 |
出版者 | MDPI |
WOS记录号 | WOS:000529890200196 |
资助机构 | National Key Research and Development Program of China ; National Natural Science Foundation of China |
源URL | [http://ir.ipe.ac.cn/handle/122111/40575] |
专题 | 中国科学院过程工程研究所 |
通讯作者 | Li, Zhimin; Li, Jian-Jun |
作者单位 | 1.Capital Normal Univ, Coll Life Sci, Beijing 10048, Peoples R China 2.Chinese Acad Sci, Natl Engn Res Ctr Biotechnol Beijing, Key Lab Biopharmaceut Prod & Formulat Engn, Natl Key Lab Biochem Engn,PLA,Inst Proc Engn, Beijing 100190, Peoples R China 3.Sichuan Normal Univ, Coll Life Sci, Chengdu 610101, Peoples R China 4.Jiangxi Agr Univ, Coll Biosci & Bioengn, Nanchang 330045, Jiangxi, Peoples R China |
推荐引用方式 GB/T 7714 | Xu, Hua,Zhang, Pengfei,Zhang, Yuchen,et al. Overexpression and Biochemical Characterization of an Endo-alpha-1,4-polygalacturonase from Aspergillus nidulans in Pichia pastoris[J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,2020,21(6):12. |
APA | Xu, Hua.,Zhang, Pengfei.,Zhang, Yuchen.,Liu, Zebin.,Zhang, Xuebing.,...&Du, Yuguang.(2020).Overexpression and Biochemical Characterization of an Endo-alpha-1,4-polygalacturonase from Aspergillus nidulans in Pichia pastoris.INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,21(6),12. |
MLA | Xu, Hua,et al."Overexpression and Biochemical Characterization of an Endo-alpha-1,4-polygalacturonase from Aspergillus nidulans in Pichia pastoris".INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 21.6(2020):12. |
入库方式: OAI收割
来源:过程工程研究所
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