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Identification of (R)-selective ω-aminotransferases by exploring evolutionary sequence space.
Eun-Mi Kim1, Joon Ho Park2, Byung-Gee Kim3
1School of Chemical and Biological Engineering, Seoul National University, Seoul 08826, Republic of Korea; AmorePacific R&D Center, Giheung-Gu, Yongin-Si, Gyeonggi-Do 17074, Republic of Korea.
Enzyme and Microbial Technology
|January 10, 2018
Summary
Researchers identified novel (R)-selective ω-aminotransferases (R-ωATs) using sequence profile analysis. This method efficiently predicts new R-ωATs, expanding the known enzyme families.
Area of Science:
- Biocatalysis
- Enzyme Engineering
- Computational Biology
Background:
- Several (R)-selective ω-aminotransferases (R-ωATs) are known, but new variants with distinct sequences are expected.
- R-ωATs are generally considered variants of aminotransferase group III.
Purpose of the Study:
- To predict and identify novel R-ωATs with different sequence characteristics.
- To develop an efficient method for discovering new R-ωATs.
Main Methods:
- Scoring sequences in the RefSeq database using family profiles of branched-chain amino acid aminotransferase (BCAT) and d-alanine aminotransferase (DAT).
- Plotting sequences on a two-dimensional score space based on BCAT and DAT profile analysis scores.
- Selecting candidate sequences with similar scores in both BCAT and DAT profiles.
Main Results:
- Putative R-ωATs were identified from Saccharopolyspora erythraea (R-ωAT_Sery), Bacillus cellulosilyticus (R-ωAT_Bcel), and Bacillus thuringiensis (R-ωAT_Bthu).
- Experimental validation confirmed R-ωAT activity for the selected candidates.
- R-ωAT_Sery exhibited DAT activity, while R-ωAT_Bcel and R-ωAT_Bthu showed BCAT activity.
Conclusions:
- Simple family profile analysis and exploration of evolutionary sequence space can efficiently identify potential R-ωATs.
- This approach expands the discovery of R-ωATs beyond previously known sequence characteristics.
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