薰衣草CuAO基因家族特征及LaCuAO1降解生物胺功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2025-0525

摘要/Abstract

摘要：

目的探究薰衣草中CuAO基因家族分子特征及LaCuAO1降解生物胺的功能，为解析铜胺氧化酶（CuAO）在植物代谢调控与生物胺降解中的作用提供参考。方法采用生物信息学方法从薰衣草（Lavandula angustifolia）全基因组数据中鉴定并筛选LaCuAO基因，克隆该基因并通过密码子优化后分别构建重组大肠杆菌菌株，利用高效液相色谱（HPLC）评估其对乙醇胺和组胺的降解效率。结果从薰衣草基因组中鉴定到LaCuAO家族19个成员，其编码蛋白分子量为39.8‒87.4 kD，外显子数目4‒12个，可分为3个独立进化分支。基因表达水平分析显示，该家族基因具有显著的组织表达特异性，其中La15G00866和La10G00763在根、茎、叶等多个组织中呈组成型高表达特征，而La05G00442仅在花蕾和花萼中特异性高表达。启动子顺式元件分析显示，启动子区域富含光响应元件及脱落酸、低温等胁迫响应元件。通过密码子优化构建的大肠杆菌工程菌optLaCuAO1（La05G00442），对乙醇胺和组胺的降解效率从野生型50.0%和92.1%分别提升至62.5%和93.2%，证实其在生物胺降解中的高效性。结论揭示薰衣草LaCuAO基因家族成员的结构和表达特征，明确重组LaCuAO1菌株对乙醇胺和组胺的高效降解能力，为解析薰衣草发育机制及开发生物胺降解新方法提供依据和参考。

关键词: 薰衣草, 铜胺氧化酶, 基因家族特征, 密码子优化, 原核表达, 基因功能, 生物胺降解

Abstract:

Objective This study aims to investigate the molecular characteristics of the CuAO gene family in lavender (Lavandula angustifolia) and the degradation function of LaCuAO1 in degrading bioamines, providing a reference for understanding the role of copper amine oxidase (CuAO) in plant metabolic regulation and biogenic amine degradation. Method Bioinformatics methods were used to identify the LaCuAO gene and screen from the genome data of lavender. The gene was cloned and then the recombinant Escherichia coli strains were constructed after optimizing the codons. High-performance liquid chromatography (HPLC) was to evaluate the degradation efficiency of ethanolamine and histamine. Result The 19 members of the LaCuAO family were identified from the lavender genome. The molecular weights of their encoded protein were of 39.8-87.4 kD, with 4-12 exons, and can be divided into 3 independent evolutionary branches. The analysis of gene expressions showed that the members of this family had significant tissue-specific expression characteristics. Among them, La15G00866 and La10G00763 showed constitutive high expression in multiple tissues such as roots, stems, and leaves, while La05G00442 was specifically and highly expressed only in flower buds and calyxes. The analysis of the promoter cis-acting elements showed that the promoter region was rich in light response elements and abscisic acid, low temperature, and other stress response elements. The engineered E. coli strain optLaCuAO1 (La05G00442) constructed through codon optimization has improved the degradation efficiency of ethanolamine and histamine from the wild type’s 50.0% and 92.1% to 62.5% and 93.2%, confirming its high efficiency in bioamine degradation. Conclusion The structural and expression characteristics of the LaCuAO gene family members in lavender are revealed, and the efficient degradation ability of the recombinant LaCuAO1 strain for ethanolamine and histamine is clarified. This provides a basis and reference for understanding the development mechanism of lavender and developing new methods for bioamine degradation.

Key words: Lavandula angustifolia, copper amine oxidase (CuAO), gene family characterization, codon optimization, prokaryotic expression, gene function, bioamine degradation

贺启琛, 杨扬, 阿丽亚·外力, 唐新月, 李忠喜, 陈永坤, 陈凌娜. 薰衣草CuAO基因家族特征及LaCuAO1降解生物胺功能研究[J]. 生物技术通报, 2026, 42(1): 114-124.

HE Qi-chen, YANG Yang, ALIYA Waili, TANG Xin-yue, LI Zhong-xi, CHEN Yong-kun, CHEN Ling-na. Investigation into the Family Characteristics of the Lavender Copper Amine Oxidase Gene and the Role of LaCuAO1 in Bioamine Degradation[J]. Biotechnology Bulletin, 2026, 42(1): 114-124.

图/表 11

图1 薰衣草不同组织图示

Fig. 1 Illustrations of different tissues of lavender

表1 相关生物信息学软件及网站

Table 1 Relevant bioinformatics software and websites

在线生物学软件 Online biological software	网站地址 Website address
Phytozome v13数据库	https：//phytozome-next.jgi.doe.gov/
CDD	https：//www.ncbi.nlm.nih.gov/cdd
HMMER	http：//www.hmmer.org/
GSDS	http：//gsds.gao-lab.org/
ExPASy	https：//web.expasy.org/protparam/
SOPMA	https：//npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl page=npsa sopma
WoLFPSORT	http：//wolfpsort.seq.cbrc.jp
iTOL	https：//itol.embl.de/
PlantCARE	http：//bioinformatics.psb.ugent.be/wetools/plantcare/html/

表2 RT-qPCR引物序列

Table 2 RT-qPCR primer sequences

基因名称 Gene ID	引物序列 Primer sequence （5′‒3′）	基因长度 Product length （bp）
La05G00442	F： GGAGTGGACAGCTTACAGGC	187
La05G00442	R： CCTTCACCGACACGTGGATT
La05G01835	F： GTGTAGGCGAAGGATTGGCT	181
La05G01835	R： GCCGGAGAGCAGTTGAAGAA
La25G00173	F： GTCGGTCGCCAACTACGATT	188
La25G00173	R： CGTGGATAACACCGACGACA
La15G00866	F： GGATCCTTCCGAGTTCCACG	171
La15G00866	R： TTGTACGGCGTCACCCAAAT
La00G03915	F： CTCTCTGGAGCAACCCGAAG	194
La00G03915	R： GCACAAACAGCTCGGAAGTG
La10G00763	F： AGGGCCTAGCTTTCGGATCA	180
La10G00763	R： TCTCCATAGGGCACGACCAT
LaActin	F：GGTCGTACAACTGGTATTGTTC R：TCCAGCAGCTTCCATTCCGATCA	139

图2 LaCuAO家族基因结构分析

Fig. 2 Gene structure analysis of LaCuAO family

表3 LaCuAO家族成员的理化性质

Table 3 Physicochemical properties of LaCuAO family members

基因ID号 Gene ID	氨基酸数 Number of amino acids	分子量 Molecular weight （Da）	理论等电点 Theoretical isoelectric point	不稳定系数 Stability coefficient	脂溶指数 Lipid solubility index	平均亲水值 Average hydrophilicity	亚细胞定位 Subcellular localization
La03G02650	784	86 774.78	6.35	42.95	79.69	-0.311	叶绿体
La05G00442	785	86 936.93	6.42	43.81	78.10	-0.330	叶绿体
La05G01835	785	87 387.54	6.55	43.60	77.49	-0.340	过氧化物酶体
La10G00763	707	78 669.53	7.64	43.69	80.82	-0.346	叶绿体
La03G03013	785	86 486.22	6.31	41.75	86.43	-0.206	叶绿体
La01G00448	665	75 822.56	5.46	41.92	81.29	-0.356	叶绿体
La22G00998	654	74 774.69	5.22	44.80	82.81	-0.315	液泡
La25G00172	714	80 710.59	5.73	36.17	83.94	-0.350	叶绿体基质
La00G03916	713	80 614.59	5.83	36.36	84.60	-0.332	叶绿体基质
La25G00173	726	81 549.49	5.77	42.39	80.66	-0.325	液泡
La15G00866	716	80 817.15	5.99	35.11	82.65	-0.300	液泡
La00G03915	726	81 556.54	5.74	42.11	80.66	-0.322	液泡
La25G00209	407	46 128.07	5.41	36.77	74.25	-0.318	线粒体
La00G03795	638	72 196.13	5.84	37.99	81.10	-0.343	细胞质
La07G01349	640	72 428.37	8.46	39.48	82.62	-0.283	液泡
La02G00655	528	59 545.29	5.18	39.84	76.17	-0.412	线粒体
La00G05435	638	72 288.29	8.52	40.93	83.03	-0.280	液泡
La17G01323	638	72 288.29	8.52	40.93	83.03	-0.280	液泡
La00G03531	352	39 783.90	5.59	32.69	75.82	-0.400	细胞质

图3 LaCuAO家族基因的共线性和系统进化分析★为薰衣草CuAO；▲为毛果杨CuAO；■为番茄CuAO；●为拟南芥CuAO

Fig. 3 Collinearity and phylogenetic analysis of LaCuAO family genes★ indicates Lavandula angustifolia CuAO. ▲ indicates Populus trichocarpa CuAO. ■ indicates Solanum lycopersicum CuAO. ● indicates Arabidopsis thaliana CuAO

图4 LaCuAO家族基因的启动子顺式调控元件分析

Fig. 4 Analysis of promoters cis-regulatory elements of LaCuAO family genes

图5 LaCuAO家族基因在薰衣草不同组织的表达TP：花瓣；TL：叶片；TS：茎；TB：花蕾；TC1：新鲜花萼；TC2：成熟花萼

Fig. 5 Expressions of LaCuAO family genes in different tissues of lavenderTP: Petal. TL: Leaf. TS: Stem. TB: Flower Bud. TC1: Fresh calyx. TC2: Mature calyx

图6 LaCuAO基因不同处理下薰衣草花蕾（A）和叶片（B）中的表达模式* P<0.05， ** P<0.01

Fig. 6 Expression patterns of LaCuAO genes in lavender buds (A) and leaves (B) under different treatments

图7 SDS-PAGE凝胶电泳图A：离心后上清液中的蛋白条带；B：沉淀中的蛋白条带；M：Marker；1‒2：优化前，优化后空白对照（未诱导）；3‒14为诱导后2‒12 h每隔2 h取样的蛋白表达

Fig. 7 SDS-PAGE gel electrophoresis patternA: Protein bands of the supernatant after centrifugation. B: Protein bands of precipitation. M: Marker. 1‒2: Blank control before and optimization (not induced). 3‒14 indicate the protein expressions sampled every 2 h at 2‒12 h after induction

图8 optLaCuAO1对乙醇胺和组胺的HPLC降解分析A：乙醇胺标准品；B：密码子优化前对乙醇胺的降解；C：密码子优化后对乙醇胺的降解；D：组胺标准品；E：密码子优化前对组胺的降解；F：密码子优化后对组胺的降解

Fig. 8 HPLC degradation analysis of ethanolamine and histamine by optLaCuAO1A: Ethanolamine reference standards. B: Degradation of ethanolamine before codon optimization. C: Degradation of ethanolamine after codon optimization. D: Histamine reference standards. E: Degradation of histamine before codon optimization. F: Degradation of histamine after codon optimization

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