Preparation of β-Casein Based on Escherichia coli Expression System
The β-casein of bovine milk contains a variety of variants, among which A1-β-casein (A1) and A2-β-casein (A2) are the two most common variants. Because of only few differences between with A1 and A2 in amino acid sequence, it is difficult to preparation of A1 and A2 proteins with higher purity by co...
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| Format: | Article |
| Language: | zho |
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The editorial department of Science and Technology of Food Industry
2023-08-01
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| Series: | Shipin gongye ke-ji |
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| Online Access: | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022090232 |
| _version_ | 1827861253735841792 |
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| author | Zhengwen AI |
| author_facet | Zhengwen AI |
| author_sort | Zhengwen AI |
| collection | DOAJ |
| description | The β-casein of bovine milk contains a variety of variants, among which A1-β-casein (A1) and A2-β-casein (A2) are the two most common variants. Because of only few differences between with A1 and A2 in amino acid sequence, it is difficult to preparation of A1 and A2 proteins with higher purity by conventional separation and purification methods. In this study, recombinant plasmid of pET28a（+）-CSN2-A1 and pET28a （+）-CSN2-A2, which contained the target genes of CSN2-A1 and CSN2-A2 were constructed by molecular biological methods, respectively. Then two recombinant vectors were introduced into Escherichia coli BL21 for induced expression and purification, respectively. The results showed that abundant of proteins could be obtained by induced expression at 0.2 mmol/L IPTG at 37 ℃ for 4 h. However, the results of SDS-PAGE showed that the target proteins were expressed in the form of inclusion bodies, and existed in the pellet after cell disruption. In addition, the purity of more than 90% (SDS-PAGE) of the A1 and A2 recombinant proteins were obtained by several processes, which contained dissolution, nickel affinity chromatography, renaturation and qualification. Thereby, it could provide a new way for the preparation of A1 and A2 proteins. |
| first_indexed | 2024-03-12T13:35:18Z |
| format | Article |
| id | doaj.art-437efadf583049e69a2b4c47041fcedc |
| institution | Directory Open Access Journal |
| issn | 1002-0306 |
| language | zho |
| last_indexed | 2024-03-12T13:35:18Z |
| publishDate | 2023-08-01 |
| publisher | The editorial department of Science and Technology of Food Industry |
| record_format | Article |
| series | Shipin gongye ke-ji |
| spelling | doaj.art-437efadf583049e69a2b4c47041fcedc2023-08-24T06:00:01ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062023-08-01441613113810.13386/j.issn1002-0306.20220902322022090232-16Preparation of β-Casein Based on Escherichia coli Expression SystemZhengwen AI0State Key Laboratory of Dairy Biotechnology, Shanghai Engineering Research Center of Dairy Biotechnology, Bright Dairy & Food Co., Ltd., Dairy Research Institute, Shanghai 200436, ChinaThe β-casein of bovine milk contains a variety of variants, among which A1-β-casein (A1) and A2-β-casein (A2) are the two most common variants. Because of only few differences between with A1 and A2 in amino acid sequence, it is difficult to preparation of A1 and A2 proteins with higher purity by conventional separation and purification methods. In this study, recombinant plasmid of pET28a（+）-CSN2-A1 and pET28a （+）-CSN2-A2, which contained the target genes of CSN2-A1 and CSN2-A2 were constructed by molecular biological methods, respectively. Then two recombinant vectors were introduced into Escherichia coli BL21 for induced expression and purification, respectively. The results showed that abundant of proteins could be obtained by induced expression at 0.2 mmol/L IPTG at 37 ℃ for 4 h. However, the results of SDS-PAGE showed that the target proteins were expressed in the form of inclusion bodies, and existed in the pellet after cell disruption. In addition, the purity of more than 90% (SDS-PAGE) of the A1 and A2 recombinant proteins were obtained by several processes, which contained dissolution, nickel affinity chromatography, renaturation and qualification. Thereby, it could provide a new way for the preparation of A1 and A2 proteins.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022090232β-caseinescherichia coliprokaryotic expressioncsn2-a1csn2-a2 |
| spellingShingle | Zhengwen AI Preparation of β-Casein Based on Escherichia coli Expression System Shipin gongye ke-ji β-casein escherichia coli prokaryotic expression csn2-a1 csn2-a2 |
| title | Preparation of β-Casein Based on Escherichia coli Expression System |
| title_full | Preparation of β-Casein Based on Escherichia coli Expression System |
| title_fullStr | Preparation of β-Casein Based on Escherichia coli Expression System |
| title_full_unstemmed | Preparation of β-Casein Based on Escherichia coli Expression System |
| title_short | Preparation of β-Casein Based on Escherichia coli Expression System |
| title_sort | preparation of β casein based on escherichia coli expression system |
| topic | β-casein escherichia coli prokaryotic expression csn2-a1 csn2-a2 |
| url | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022090232 |
| work_keys_str_mv | AT zhengwenai preparationofbcaseinbasedonescherichiacoliexpressionsystem |