普通烟草NtASAT2的结构预测、克隆表达及功能验证

韩丽; 董滋强; 王丽娇; 李文钦; 王晨辉; 肖成志; 毛多斌

Article Contents

HAN Li, DONG Ziqiang, WANG Lijiao, et al. Structural prediction, cloning expression and functional verification of NtASAT2 from Nicotiana tabacum L.[J]. Journal of Light Industry.

Citation: HAN Li, DONG Ziqiang, WANG Lijiao, et al. Structural prediction, cloning expression and functional verification of NtASAT2 from Nicotiana tabacum L.[J]. Journal of Light Industry. shu

Structural prediction, cloning expression and functional verification of NtASAT2 from Nicotiana tabacum L.

HAN Li ^1,2 ,
DONG Ziqiang ¹ ,
WANG Lijiao ¹ ,
LI Wenqin ¹ ,
WANG Chenhui ¹ ,
XIAO Chengzhi ¹ ,
MAO Duobin ^1,2

1. College of Food and Bioengineering, Zhengzhou University of Light Industry, Zhengzhou 450001, China;
2. Key Laboratory of Biotechnology in Tobacco Industry, Zhengzhou University of Light Industry, Zhengzhou 450001, China

Received Date: 2024-05-10
Accepted Date: 2024-05-27
Available Online: 2025-04-16

Abstract

To obtain functional acyl sugar acyltransferase NtASAT2 of Nicotiana tabacum L., bioinformatic methods were applied to analyze and predict the sequences and structure of NtASAT2. The gene of NtASAT2 was cloned, prokaryotic expressed, and then purified. T prokaryotic expressed, and then purified. The function of the recombinant protein was verified through enzymatic catalysis. The results showed that in the secondary structure of NtASAT2, α-helices and random coils account for a large proportion with 39. 04% and 41. 13%, respectively. The amino acid sequence of NtASAT2 was highly similar to that of sticky tobacco NacASAT2. The content of soluble NtASAT2 in the recombinant protein expressed by prokaryotic cell BL21 (DE3) was relatively low, and the affinity of NtASAT2 to nickel column was weak. After purification, only a small amount of the target protein was obtained. In enzymatic reaction systems containing substrates, NtASAT2 showed enzyme activity and catalyzed the formation of sucrose diesters. The research results could provide a theoretical basis for the application of acyl sugar acyltransferase in enzymatic synthesis of sucrose esters.
- Nicotiana tabacum L.,
- acylsugar acyltransferase,
- sucrose ester,
- cloning,
- expression and purification
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沈阳化工大学材料科学与工程学院沈阳 110142

Structural prediction, cloning expression and functional verification of NtASAT2 from Nicotiana tabacum L.

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