DOI: 10.12187/2025.04.008
Structural prediction,cloning expression,and functional verification of NtASAT2 from Nicotiana tabacum L.
HAN Li
DONG Ziqiang
WANG Lijiao
LI Wenqin
WANG Chenhui
XIAO Chengzhi
MAO Duobin
Abstract：To obtain active acylsugar acyltransferase NtASAT2 from Nicotiana tabacum L.,bioinformatic methods were employed to analyze and predict its sequence and structure.The NtASAT2 gene was cloned,prokaryotically expressed in Escherichia coli BL21(DE3),and subsequently purified.The function of the recombinant protein was characterized through enzymatic catalysis.The results showed that in the secondary structure of NtASAT2,α-helices and random coils accounted for 39.04%and 41.13%of the total structure,respectively.The amino acid sequence of NtASAT2 was highly similar to that of Nicotiana glutinosa NacASAT2.The solubility of NtASAT2 in the recombinant protein expressed in E.coli BL21(DE3)was low,and NtASAT2 exhibited weak binding affinity for the nickel column.Consequently,only a small amount of the target protein was successfully purified.In enzymatic reaction systems supplemented with substrates,NtASAT2 exhibited enzymatic activity and catalyzed the production of sucrose diesters.These findings provided a theoretical basis for the application of NtASAT2 in the enzymatic catalysis of sucrose ester synthesis.
Keywords：Nicotiana tabacum L.acylsugar acyltransferasesucrose estercloningexpression and purification
Publication Date：2025-08-15
Online Publishing Date：2025-08-19(First online date of this platform, not the publication date of the document)
Pages：8( 69-76 )