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来自马铃薯的主要体内甾体生物碱葡萄糖基转移酶。

The primary in vivo steroidal alkaloid glucosyltransferase from potato.

作者信息

McCue Kent F, Allen Paul V, Shepherd Louise V T, Blake Alison, Whitworth Jonathan, Maccree M Malendia, Rockhold David R, Stewart Derek, Davies Howard V, Belknap William R

机构信息

USDA, Agricultural Research Service, Crop Improvement and Utilization Research Unit, Western Regional Research Center, 800 Buchanan St., Albany, CA 94710-1105, USA.

出版信息

Phytochemistry. 2006 Aug;67(15):1590-7. doi: 10.1016/j.phytochem.2005.09.037. Epub 2005 Nov 18.

Abstract

To provide tools for breeders to control the steroidal glycoalkaloid (SGA) pathway in potato, we have investigated the steroidal alkaloid glycosyltransferase (Sgt) gene family. The committed step in the SGA pathway is the glycosylation of solanidine by either UDP-glucose or UDP-galactose leading to alpha-chaconine or alpha-solanine, respectively. The Sgt2 gene was identified by deduced protein sequence homology to the previously identified Sgt1 gene. SGT1 has glucosyltransferase activity in vitro, but in vivo serves as the UDP-galactose:solanidine galactosyltransferase. Two alleles of the Sgt2 gene were isolated and its function was established with antisense transgenic lines and in vitro assays of recombinant protein. In tubers of transgenic potato (Solanum tuberosum) cvs. Lenape and Desirée expressing an antisense Sgt2 gene construct, accumulation of alpha-solanine was increased and alpha-chaconine was reduced. Studies with recombinant SGT2 protein purified from yeast show that SGT2 glycosylation activity is highly specific for UDP-glucose as a sugar donor. This data establishes the function of the gene product (SGT2), as the primary UDP-glucose:solanidine glucosyltransferase in vivo.

摘要

为了给育种者提供控制马铃薯中甾体糖苷生物碱(SGA)途径的工具,我们研究了甾体生物碱糖基转移酶(Sgt)基因家族。SGA途径中的关键步骤是茄啶分别被UDP-葡萄糖或UDP-半乳糖糖基化,分别生成α-查茄碱或α-茄碱。通过推导的蛋白质序列与先前鉴定的Sgt1基因的同源性鉴定出Sgt2基因。SGT1在体外具有葡萄糖基转移酶活性,但在体内作为UDP-半乳糖:茄啶半乳糖基转移酶。分离出Sgt2基因的两个等位基因,并通过反义转基因系和重组蛋白的体外测定确定了其功能。在表达反义Sgt2基因构建体的转基因马铃薯(Solanum tuberosum)品种Lenape和Desirée的块茎中,α-茄碱的积累增加,α-查茄碱减少。对从酵母中纯化的重组SGT2蛋白的研究表明,SGT2糖基化活性对作为糖供体的UDP-葡萄糖具有高度特异性。这些数据确定了基因产物(SGT2)在体内作为主要的UDP-葡萄糖:茄啶葡萄糖基转移酶的功能。

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