Expression，Purification and Crystallization of a Glycoside Hydrolase Family 5 Endoglucanase（AnCel5A） from Aspergillus niger

doi:10.3969/j.issn.1673-1689.2019.02.012

Home > Archive>Volume 38, Issue 2, 2019 >81-86. DOI:10.3969/j.issn.1673-1689.2019.02.012

Expression，Purification and Crystallization of a Glycoside Hydrolase Family 5 Endoglucanase（AnCel5A） from Aspergillus niger
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                        10.3969/j.issn.1673-1689.2019.02.012
                    
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The β-1，4-endoglucanase from Aspergillu sniger（AnCel5A） is a cellulase which is classified to glycoside hydrolases subfamily （GH） 5. It can hydrolyze the amorphous region of cellulose long chains by endo-action，which makes it one of the key enzymes in cellulose hydrolysis.This research aims at determination the crystal structure of AnCel5A by screening and optimization its protein crystals. The signal peptide truncated AnCel5A is successfully expressed in P. pastoris X33. Purified proteins are obtained by using ion-exchange and hydrophobic chromatography. The crystal screen of AnCel5A is completed by using sitting-drop vapor diffusion method and screen kits bought from the Hampton Research company. Initial crystals are found in the reservoir solution of cryoI kit （A6）. After further optimization，better crystals are obtained. The crystals diffracted to 1.8 ? and dataset is collected at beam line BL15A1 of the National Synchrotron Radiation Research Center （NSRRC，Hsinchu，Taiwan）.

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YAN Junjie, LI Yujie, ZHENG Yingying, CHEN Chunchi, GUO Reyting, LI Huazhong, LIU Weidong. Expression，Purification and Crystallization of a Glycoside Hydrolase Family 5 Endoglucanase（AnCel5A） from Aspergillus niger[J]. Journal of Food Science and Biotechnology,2019,38(2):81-86.

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Online: March 28,2019
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