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当前位置：农业与食品科学 > 2024年 4卷 (3)期

Open Access Article

Journal of Agriculture and Food Science. 2024; 4: (3) ; 17-23 ; DOI: 10.12208/j.jafs.20241016.

Bioinformatics Analysis of protein Encoded by Norcoclaurine synthase in Piper nigrum
胡椒去甲乌药碱合酶基因编码蛋白质的生物信息学分析

作者： 邢超^1,2, 姜洁^1,2, 耿健强^1,2, 王丹^1,2, 宋丽萍^1,2, 毛婷³ *

1 北京市食品检验研究院（北京市食品安全监控和风险评估中心）北京

2 国家市场监管重点实验室（食品安全重大综合保障关键技术）北京

3 北京市检验检测认证中心北京

*通讯作者：毛婷,单位：北京市检验检测认证中心北京；

重大赛事食源性兴奋剂检验和防控关键技术研究及示范应用创新团队项目

发布时间: 2024-09-29 总浏览量: 556

PDF 全文下载引用本文万方数据（WANFANG DATA）

摘要

胡椒中含有一种β受体激动剂类物质—去甲乌药碱，具有误食而发生兴奋剂阳性事件的风险。本研究基于胡椒基因组blast，发现胡椒中存在三个去甲乌药碱合酶（PnNCS）基因。首先利用生物信息学方法对三个PnNCS的氨基酸序列进行了理化性质分析。随后，PnNCS的系统发育地位分析结果表明去甲乌药碱合酶分为八个进化分支，胡椒的三种去甲乌药碱合酶分属在不同的两个进化分支中。保守基序分析结果显示共具有三种保守基序，分别为：YGQAFVVSEDQKLDWGMLFLJTLPVELRNMRFWPTNPPSF、FTKVDNEKMVKETEVVEGGYLDFGFTLYRVRFEI以及KLHLACKEWGFFQLINHGVPE。胡椒NCS：APF31815.1和AS075045.1具有一致的保守基序（FTKVDNEKMVKETEVVEGGYLDFGFTLYRVRFEI），胡椒NCS：WDS62667.1无明确保守基序。修饰位点分析结果显示胡椒NCS蛋白具有多个磷酸化、糖基化以及氧化还原修饰位点。顺式作用元件分析结果显示NCS的启动区含有应答环境胁迫与多种植物激素的顺式作用元件。本研究提供了胡椒NCS氨基酸的基础信息，为后续探究遗传特性提供了参考。

关键词： 胡椒；去甲乌药碱合酶；生物信息学预测

Abstract

There is a β-agonist substance in pepper - noraconine, which has the risk of positive stimulant events due to ingestion. In this study, based on the pepper genome blast, we found three normonine synthase (PnNCS) genes in pepper. Firstly, the amino acid sequences of three PnNCS were analyzed by bioinformatics method. Subsequently, phylogenetic status analysis of PnNCS showed that noraconitine synthases were divided into eight evolutionary clades, and the three noraconitine synthases of pepper belonged to two different evolutionary clades. The results of conservative motifs analysis showed that there were three kinds of conservative motifs, which were: YGQAFVVSEDQKLDWGMLFLJTLPVELRNMRFWPTNPPSF, FTKVDNEKMVKETEVVEGGYLDFGFTLYR VRFEI and KLHLACKEWGFFQLINHGVPE. Pepper replication: APF31815.1 and AS075045.1 conservative base with a consistent sequence (FTKVDNEKMVKETEVVEGGYLDFGFTLYRVRFEI), pepper replication: WDS62667.1 no clearly conservative motif. The analysis of modification sites showed that the NCS protein had multiple phosphorylation, glycosylation and REDOX modification sites. The analysis of cis-acting elements showed that the initiation region of NCS contains cis-acting elements in response to environmental stress and various plant hormones. This study provided the basic information of NCS amino acids of pepper, and provided a reference for further investigation of genetic characteristics.

Key words： Pepper; Noraconine synthase; Bioinformatics prediction

参考文献 References

[1] Samanani N, Liscombe DK, Facchini PJ. Molecular cloning and characterization of norcoclaurine synthase, an enzyme catalyzing the first committed step in benzylisoquinoline alkaloid biosynthesis. Plant J 2004, 40(2):302-313.

[2] Samanani N and Facchini PJ. Isolation and partial characterization of norcoclaurine synthase, the first committed step in benzylisoquinoline alkaloid biosynthesis, from opium poppy. Planta 2001, 213(6):898-906.

[3] Samanani N and Facchini PJ. Purification and characterization of norcoclaurine synthase. The first committed enzyme in benzylisoquinoline alkaloid biosynthesis in plants. J Biol Chem 2002, 277(37):33878-33883.

[4] Minami H, Dubouzet E, Iwasa K, et al. Functional analysis of norcoclaurine synthase in Coptis japonica. J Biol Chem 2007, 282(9):6274-6282.

[5] Luk LY, Bunn S, Liscombe DK, et al. Mechanistic studies on norcoclaurine synthase of benzylisoquinoline alkaloid biosynthesis: an enzymatic Pictet-Spengler reaction. Biochemistry 2007, 46(35):10153-10161.

[6] Berkner H, Engelhorn J, Liscombe DK, et al. High-yield expression and purification of isotopically labeled norcoclaurine synthase, a Bet v 1-homologous enzyme, from Thalictrum flavum for NMR studies. Protein Expr Purif 2007, 56(2):197-204.

[7] Lee SK and Keasling JD. Heterologous protein production in Escherichia coli using the propionate-inducible pPro system by conventional and auto-induction methods. Protein Expr Purif 2008, 61(2):197-203.

[8] Berkner H, Schweimer K, Matecko I. et al. Conformation, catalytic site, and enzymatic mechanism of the PR10 allergen-related enzyme norcoclaurine synthase. Biochem J 2008, 413(2):281-290.

[9] Ilari A, Franceschini S, Bonamore A. et al., Structural basis of enzymatic (S)-norcoclaurine biosynthesis. J Biol Chem 2009, 284(2):897-904.

[10] Pasquo A, Bonamore A, Franceschini S. et al. Cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from Thalictrum flavum. Acta Crystallogr Sect F Struct Biol Cryst Commun 2008, 64(Pt 4):281-283.

[11] Lee EJ and Facchini P. Norcoclaurine synthase is a member of the pathogenesis-related 10/Bet v1 protein family. Plant Cell 2010, 22(10):3489-3503.

[12] Li J, Lee EJ, Chang L, et al. Genes encoding norcoclaurine synthase occur as tandem fusions in the Papaveraceae. Sci Rep 2016, 6:39256.

[13] Vimolmangkang S, Deng X, Owiti A, et al. Evolutionary origin of the NCSI gene subfamily encoding norcoclaurine synthase is associated with the biosynthesis of benzylisoquinoline alkaloids in plants. Sci Rep 2016, 6:26323.

[14] Yang M, Zhu L, Li L, et al. Digital Gene Expression Analysis Provides Insight into the Transcript Profile of the Genes Involved in Aporphine Alkaloid Biosynthesis in Lotus (Nelumbo nucifera). Front Plant Sci 2017, 8:80.

[15] Chen DX, Pan Y, Wang Y, et al.The chromosome-level reference genome of Coptis chinensis provides insights into genomic evolution and berberine biosynthesis. Hortic Res 2021, 8(1):121.

[16] Hao DC, Li P, Xiao PG, et al. Dissection of full-length transcriptome and metabolome of Dichocarpum (Ranunculaceae): implications in evolution of specialized metabolism of Ranunculales medicinal plants. PeerJ 2021, 9:e12428.

[17] 冯飞雪,吕瑞华,李依民等.延胡索去甲乌药碱合成酶基因CyNCS1克隆及其表达模式分析[J/OL].西北农林科技大学学报(自然科学版),2024(03):1-9.

[18] Dastmalchi M. Elusive partners: a review of the auxiliary proteins guiding metabolic flux in flavonoid biosynthesis. Plant J 2021, 108(2):314-329.

[19] 郝晓燕，丁璇，赵准，胡文冉.陆地棉GhTCP4基因编码蛋白质生物信息学分析[J/OL].分子植物育种. https://link.cnki.net/urlid/46.1068.S.20240513.1736.006.

[20] Vu LD, Gevaert K, De Smet I. Protein Language: Post-Translational Modifications Talking to Each Other. Trends Plant Sci. 2018 Dec;23(12):1068-1080.

[21] Patwardhan A, Cheng N, Trejo J. Post-Translational Modifications of G Protein-Coupled Receptors Control Cellular Signaling Dynamics in Space and Time. Pharmacol Rev. 2021 Jan;73(1):120-151.

引用本文

邢超, 姜洁, 耿健强, 王丹, 宋丽萍, 毛婷, 胡椒去甲乌药碱合酶基因编码蛋白质的生物信息学分析[J]. 农业与食品科学, 2024; 4: (3) : 17-23.

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