轻工学报2025，Vol.40Issue(4)：69-76,8.DOI:10.12187/2025.04.008
普通烟草NtASAT2的结构预测、克隆表达及功能验证
Structural prediction,cloning expression,and functional verification of NtASAT2 from Nicotiana tabacum L
摘要
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.关键词
普通烟草/酰基糖酰基转移酶/蔗糖酯/克隆/表达与纯化Key words
Nicotiana tabacum L./acylsugar acyltransferase/sucrose ester/cloning/expression and purification分类
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韩丽,董滋强,王丽娇,李文钦,王晨辉,肖成志,毛多斌..普通烟草NtASAT2的结构预测、克隆表达及功能验证[J].轻工学报,2025,40(4):69-76,8.基金项目
中国烟草总公司重点研发项目(110202202006) （110202202006）
