Preliminary communication
https://doi.org/10.17113/ftb.56.01.18.5445
Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli
Yankun Yang
; The Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Jiangnan University, Ministry of Education
Guoqiang Liu
; The Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Jiangnan University, Ministry of Education
Meng Liu
; National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University
Zhonghu Bai
; National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University
Xiuxia Liu
; National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University
Xiaofeng Dai
; Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University
Wenwen Guo
; The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University
Abstract
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.
Keywords
domain antibody (dAb); Escherichia coli; heterologous protein soluble expression; linear modelling; primary structure
Hrčak ID:
197382
URI
Publication date:
30.3.2018.
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