Title |
Enhancement of the solubility of recombinant proteins by fusion with a short-disordered peptide |
Author |
Jun Ren, Suhee Hwang, Junhao Shen, Hyeongwoo Kim, Hyunjoo Kim, Jieun Kim, Soyoung Ahn, Min-gyun Kim, Seung Ho Lee, and Dokyun Na* |
Address |
Department of Biomedical Engineering, Chung-Ang University, Seoul 06974, Republic of Korea |
Bibliography |
Journal of Microbiology, 60(9),960-967, 2022,
|
DOI |
10.1007/s12275-022-2122-z
|
Key Words |
disordered short peptide, solubility, fusion tag,
recombinant proteins |
Abstract |
In protein biotechnology, large soluble fusion partners are
widely utilized for increased yield and solubility of recombinant
proteins. However, the production of additional large
fusion partners poses an additional burden to the host, leading
to a decreased protein yield. In this study, we identified
two highly disordered short peptides that were able to increase
the solubility of an artificially engineered aggregationprone
protein, GFP-GFIL4, from 0.6% to 61% (D3-DP00592)
and 46% (D4-DP01038) selected from DisProt database. For
further confirmation, the peptides were applied to two insoluble
E. coli proteins (YagA and YdiU). The peptides also
enhanced solubility from 52% to 90% (YagA) and from 27%
to 93% (YdiU). Their ability to solubilize recombinant proteins
was comparable with strong solubilizing tags, maltosebinding
protein (40 kDa) and TrxA (12 kDa), but much smaller
(< 7 kDa) in size. For practical application, the two peptides
were fused with a restriction enzyme, I-SceI, and they increased
I-SceI solubility from 24% up to 75%. The highly disordered
peptides did not affect the activity of I-SceI while I-SceI fused
with MBP or TrxA displayed no restriction activity. Despite
the small size, the highly disordered peptides were able to
solubilize recombinant proteins as efficiently as conventional
fusion tags and did not interfere with the function of recombinant
proteins. Consequently, the identified two highly disordered
peptides would have practical utility in protein biotechnology
and industry. |