Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli
It is widely accepted that features such as pI, length, molecular mass and amino acid (AA) sequence have a significant influence on protein solubility. Here, we mainly focused on AA composition and explored those that most affected the soluble expression level of human serum albumin (HSA) domain ant...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
University of Zagreb Faculty of Food Technology and Biotechnology
2018-01-01
|
| Series: | Food Technology and Biotechnology |
| Subjects: | |
| Online Access: | https://hrcak.srce.hr/file/290750 |
| _version_ | 1797207714025177088 |
|---|---|
| author | Yankun Yang Guoqiang Liu Meng Liu Zhonghu Bai Xiuxia Liu Xiaofeng Dai Wenwen Guo |
| author_facet | Yankun Yang Guoqiang Liu Meng Liu Zhonghu Bai Xiuxia Liu Xiaofeng Dai Wenwen Guo |
| author_sort | Yankun Yang |
| collection | DOAJ |
| description | It is widely accepted that features such as pI, length, molecular mass and amino acid (AA) sequence have a significant influence on protein solubility. Here, we mainly focused on AA composition and explored those that most affected the soluble expression level of human serum albumin (HSA) domain antibody (dAb). The soluble expression and sequence of 65 dAb variants were analysed using clustering and linear modelling. Certain AAs significantly affected the soluble expression level of dAb, with the specific AA combinations being (S, R, N, D, Q), (G, R, C, N, S) and (R, S, G); these combinations respectively affected the dAb expression level in the broth supernatant, the level in the pellet lysate and total soluble dAb. Among the 20 AAs, R displayed a negative influence on the soluble expression level, whereas G and S showed positive effects. A linear model was built to predict the soluble expression level from the sequence; this model had a prediction accuracy of 80 %. In summary, increasing the content of polar AAs, especially G and S, and decreasing the content of R, was helpful to improve the soluble expression level of HSA dAb. |
| first_indexed | 2024-04-24T09:27:18Z |
| format | Article |
| id | doaj.art-ed80fc4de303454398a0cafad6ee99ec |
| institution | Directory Open Access Journal |
| issn | 1330-9862 1334-2606 |
| language | English |
| last_indexed | 2024-04-24T09:27:18Z |
| publishDate | 2018-01-01 |
| publisher | University of Zagreb Faculty of Food Technology and Biotechnology |
| record_format | Article |
| series | Food Technology and Biotechnology |
| spelling | doaj.art-ed80fc4de303454398a0cafad6ee99ec2024-04-15T14:42:15ZengUniversity of Zagreb Faculty of Food Technology and BiotechnologyFood Technology and Biotechnology1330-98621334-26062018-01-0156110110910.17113/ftb.56.01.18.5445Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coliYankun Yang0Guoqiang Liu1Meng Liu2Zhonghu Bai3Xiuxia Liu4Xiaofeng Dai5Wenwen Guo6The Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Jiangnan University, Ministry of EducationThe Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Jiangnan University, Ministry of EducationNational Engineering Laboratory for Cereal Fermentation Technology, Jiangnan UniversityNational Engineering Laboratory for Cereal Fermentation Technology, Jiangnan UniversityNational Engineering Laboratory for Cereal Fermentation Technology, Jiangnan UniversityJiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan UniversityThe Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan UniversityIt is widely accepted that features such as pI, length, molecular mass and amino acid (AA) sequence have a significant influence on protein solubility. Here, we mainly focused on AA composition and explored those that most affected the soluble expression level of human serum albumin (HSA) domain antibody (dAb). The soluble expression and sequence of 65 dAb variants were analysed using clustering and linear modelling. Certain AAs significantly affected the soluble expression level of dAb, with the specific AA combinations being (S, R, N, D, Q), (G, R, C, N, S) and (R, S, G); these combinations respectively affected the dAb expression level in the broth supernatant, the level in the pellet lysate and total soluble dAb. Among the 20 AAs, R displayed a negative influence on the soluble expression level, whereas G and S showed positive effects. A linear model was built to predict the soluble expression level from the sequence; this model had a prediction accuracy of 80 %. In summary, increasing the content of polar AAs, especially G and S, and decreasing the content of R, was helpful to improve the soluble expression level of HSA dAb.https://hrcak.srce.hr/file/290750domain antibody (dAb)Escherichia coliheterologous protein soluble expressionlinear modellingprimary structure |
| spellingShingle | Yankun Yang Guoqiang Liu Meng Liu Zhonghu Bai Xiuxia Liu Xiaofeng Dai Wenwen Guo Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli Food Technology and Biotechnology domain antibody (dAb) Escherichia coli heterologous protein soluble expression linear modelling primary structure |
| title | Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli |
| title_full | Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli |
| title_fullStr | Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli |
| title_full_unstemmed | Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli |
| title_short | Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli |
| title_sort | correlation between protein primary structure and soluble expression level of hsa dab in escherichia coli |
| topic | domain antibody (dAb) Escherichia coli heterologous protein soluble expression linear modelling primary structure |
| url | https://hrcak.srce.hr/file/290750 |
| work_keys_str_mv | AT yankunyang correlationbetweenproteinprimarystructureandsolubleexpressionlevelofhsadabinescherichiacoli AT guoqiangliu correlationbetweenproteinprimarystructureandsolubleexpressionlevelofhsadabinescherichiacoli AT mengliu correlationbetweenproteinprimarystructureandsolubleexpressionlevelofhsadabinescherichiacoli AT zhonghubai correlationbetweenproteinprimarystructureandsolubleexpressionlevelofhsadabinescherichiacoli AT xiuxialiu correlationbetweenproteinprimarystructureandsolubleexpressionlevelofhsadabinescherichiacoli AT xiaofengdai correlationbetweenproteinprimarystructureandsolubleexpressionlevelofhsadabinescherichiacoli AT wenwenguo correlationbetweenproteinprimarystructureandsolubleexpressionlevelofhsadabinescherichiacoli |