Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector
At present, approximately 30% of eukaryotic proteins can be expressed in a soluble form in Escherichia coli. In this study, a pCold-SUMOa plasmid was constructed in order to express heterologous proteins fused with SUMO by a cold-shock expression vector. The human cysteine desulfurase NFS1 and a chi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-09-01
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| Series: | Biotechnology Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215017X17303041 |
| _version_ | 1817991685200674816 |
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| author | Jianghui Li Qinxia Han Tao Zhang Jing Du Qianqian Sun Yilin Pang |
| author_facet | Jianghui Li Qinxia Han Tao Zhang Jing Du Qianqian Sun Yilin Pang |
| author_sort | Jianghui Li |
| collection | DOAJ |
| description | At present, approximately 30% of eukaryotic proteins can be expressed in a soluble form in Escherichia coli. In this study, a pCold-SUMOa plasmid was constructed in order to express heterologous proteins fused with SUMO by a cold-shock expression vector. The human cysteine desulfurase NFS1 and a chimeric cysteine desulfurase namely, EH-IscS were successfully expressed in E. coli. The proteins were particularly difficult to be produced functionally, due to their readily sequestered nature. The recombinant cysteine desulfurases that were generated by pCold-SUMOa exhibited higher activity, solubility and stability compared with the well-known plasmid pCold I. In contrast to the pCold TF plasmid, the SUMO tag conferred no biological activity with regard to the conformation of the cysteine desulfurases. Furthermore, the SUMO protease 1 can efficiently recognize the tertiary structure of SUMO and cleave it. The data indicate that the pCold-SUMOa vector is a promising tool for native eukaryotic protein production. Keywords: Prokaryotic expression system, SUMO, Ulp1, Cold-shock expression vector, NFS1 |
| first_indexed | 2024-04-14T01:16:32Z |
| format | Article |
| id | doaj.art-0bef6ea68fdb4c1eac44543e802d3dcc |
| institution | Directory Open Access Journal |
| issn | 2215-017X |
| language | English |
| last_indexed | 2024-04-14T01:16:32Z |
| publishDate | 2018-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biotechnology Reports |
| spelling | doaj.art-0bef6ea68fdb4c1eac44543e802d3dcc2022-12-22T02:20:50ZengElsevierBiotechnology Reports2215-017X2018-09-0119Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vectorJianghui Li0Qinxia Han1Tao Zhang2Jing Du3Qianqian Sun4Yilin Pang5Key Laboratory of Laboratory Medicine, Ministry of Education, Zhejiang Provincial Key Laboratory of Medical Genetics, Institute of Enzyme Engineering and Medical Diagnosis, College of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, ChinaKey Laboratory of Laboratory Medicine, Ministry of Education, Zhejiang Provincial Key Laboratory of Medical Genetics, Institute of Enzyme Engineering and Medical Diagnosis, College of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, ChinaKey Laboratory of Laboratory Medicine, Ministry of Education, Zhejiang Provincial Key Laboratory of Medical Genetics, Institute of Enzyme Engineering and Medical Diagnosis, College of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, ChinaPeople's Hospital of Hangzhou Medical College, Hangzhou 310014, ChinaKey Laboratory of Laboratory Medicine, Ministry of Education, Zhejiang Provincial Key Laboratory of Medical Genetics, Institute of Enzyme Engineering and Medical Diagnosis, College of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, ChinaKey Laboratory of Laboratory Medicine, Ministry of Education, Zhejiang Provincial Key Laboratory of Medical Genetics, Institute of Enzyme Engineering and Medical Diagnosis, College of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China; Corresponding author at: College of Laboratory Medicine and Life Sciences, Wenzhou Medical University, B409 Tong De Bldg, Chashan University Town, Wenzhou, Zhejiang 325035, China.At present, approximately 30% of eukaryotic proteins can be expressed in a soluble form in Escherichia coli. In this study, a pCold-SUMOa plasmid was constructed in order to express heterologous proteins fused with SUMO by a cold-shock expression vector. The human cysteine desulfurase NFS1 and a chimeric cysteine desulfurase namely, EH-IscS were successfully expressed in E. coli. The proteins were particularly difficult to be produced functionally, due to their readily sequestered nature. The recombinant cysteine desulfurases that were generated by pCold-SUMOa exhibited higher activity, solubility and stability compared with the well-known plasmid pCold I. In contrast to the pCold TF plasmid, the SUMO tag conferred no biological activity with regard to the conformation of the cysteine desulfurases. Furthermore, the SUMO protease 1 can efficiently recognize the tertiary structure of SUMO and cleave it. The data indicate that the pCold-SUMOa vector is a promising tool for native eukaryotic protein production. Keywords: Prokaryotic expression system, SUMO, Ulp1, Cold-shock expression vector, NFS1http://www.sciencedirect.com/science/article/pii/S2215017X17303041 |
| spellingShingle | Jianghui Li Qinxia Han Tao Zhang Jing Du Qianqian Sun Yilin Pang Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector Biotechnology Reports |
| title | Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector |
| title_full | Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector |
| title_fullStr | Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector |
| title_full_unstemmed | Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector |
| title_short | Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector |
| title_sort | expression of soluble native protein in escherichia coli using a cold shock sumo tag fused expression vector |
| url | http://www.sciencedirect.com/science/article/pii/S2215017X17303041 |
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