Investigation into the Family Characteristics of the Lavender Copper Amine Oxidase Gene and the Role of LaCuAO1 in Bioamine Degradation

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

Abstract

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

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.

Figures/Tables 11

References 31

[1]	Wang ZH, Wang Y, Yang J, et al. Reliable selection and holistic stability evaluation of reference genes for rice under 22 different experimental conditions [J]. Appl Biochem Biotechnol, 2016, 179(5): 753-775.
[2]	屈海峰, 王登伟. 伊犁河谷油用薰衣草高产栽培技术 [J]. 新疆农垦科技, 2016, 39(8): 16-17.
	Qu HF, Wang DW. High-yield cultivation techniques of oil lavender in Ili valley [J]. Xinjiang Farm Res Sci Technol, 2016, 39(8): 16-17.
[3]	陈国通, 左书瑞, 曹雪琴, 等. 吹扫捕集-气相色谱-质谱法测定薰衣草香气成分 [J]. 安徽农业科学, 2019, 47(2): 191-193.
	Chen GT, Zuo SR, Cao XQ, et al. Determination of aroma components of lavender by purge and trap-gas chromatography-mass spectrometry [J]. J Anhui Agric Sci, 2019, 47(2): 191-193.
[4]	Sharma M, Grewal K, Jandrotia R, et al. Essential oils as anticancer agents: potential role in malignancies, drug delivery mechanisms, and immune system enhancement [J]. Biomed Pharmacother, 2022, 146: 112514.
[5]	Crişan I, Ona A, Vârban D, et al. Current trends for lavender (Lavandula angustifolia Mill.) crops and products with emphasis on essential oil quality [J]. Plants, 2023, 12(2): 357.
[6]	Gonzalez ME, Jasso-Robles FI, Flores-Hernández E, et al. Current status and perspectives on the role of polyamines in plant immunity [J]. Ann Appl Biol, 2021, 178(2): 244-255.
[7]	王伟, 程鑫, 崔振国, 等. 铜胺氧化酶基因PpCuAO4在桃成熟中的功能鉴定 [J]. 核农学报, 2022, 36(9): 1738-1745.
	Wang W, Cheng X, Cui ZG, et al. Function identification of copper amine oxidase gene PpCuAO4 during ripening of peach (Prunus persica L.) fruit [J]. J Nucl Agric Sci, 2022, 36(9): 1738-1745.
[8]	Yu TQ, Yin YF, Ge YH, et al. Enzymatic production of 4-hydroxyphenylacetaldehyde by oxidation of the amino group of tyramine with a recombinant primary amine oxidase [J]. Process Biochem, 2020, 92: 105-112.
[9]	Li JR, Wang YM, Dong YM, et al. The chromosome-based lavender genome provides new insights into Lamiaceae evolution and terpenoid biosynthesis [J]. Hortic Res, 2021, 8(1): 53.
[10]	Liu SP, Yao HL, Sun MF, et al. Heterologous expression and characterization of amine oxidases from Saccharopolyspora to reduce biogenic amines in huangjiu [J]. LWT, 2022, 169: 113963.
[11]	徐圆圆, 赵国春, 郝颖颖, 等. 无患子RT-qPCR内参基因的筛选与验证 [J]. 生物技术通报, 2022, 38(10): 80-89.
	Xu YY, Zhao GC, Hao YY, et al. Reference genes selection and validation for RT-qPCR in Sapindus mukorossi [J]. Biotechnol Bull, 2022, 38(10): 80-89.
[12]	岳俊齐, 约日耶提·萨力, 克拉热木·克里木江, 等. 薰衣草LaGGPPS5基因在大肠杆菌中催化萜类物质合成 [J]. 中国农业科技导报, 2023, 25(12): 85-92.
	Yue JQ, Yueriyeti S, Kelaremu K, et al. Lavender LaGGPPS5 gene catalyzing terpenoids synthesis in Escherichia coli [J]. J Agric Sci Technol, 2023, 25(12): 85-92.
[13]	陈志军, 张立坚, 池青, 等. 小麦NAC转录因子TaNAC14的原核表达和分离纯化 [J]. 生物工程学报, 2024, 40(11): 4171-4182.
	Chen ZJ, Zhang LJ, Chi Q, et al. Prokaryotic expression and purification of the transcription factor TaNAC14 in wheat (Triticum aestivum) [J]. Chin J Biotechnol, 2024, 40(11): 4171-4182.
[14]	姜维. 一株耐盐性高效生物胺降解新菌的筛选、分类鉴定及应用研究 [D]. 青岛: 中国海洋大学, 2014.
	Jiang W. Study on the screening, taxonomic analysis and application of one novel, salt-tolerant and high efficient biogenic amines degrading bacterium [D]. Qingdao: Ocean University of China, 2014.
[15]	胡昌勤, 张夏, 常祎卓. HPLC分析的全生命周期管理 [J]. 中国现代应用药学, 2024, 41(20): 2758-2767.
	Hu CQ, Zhang X, Chang YZ. Life-cycle management of HPLC analysis [J]. Chin J Mod Appl Pharm, 2024, 41(20): 2758-2767.
[16]	克拉热木·克里木江, 陈永坤, 李翠翠, 等. 薰衣草DXS基因家族的全基因组鉴定和表达分析 [J]. 基因组学与应用生物学, 2023, 42(9): 919-926.
	Kelaremu·K, Chen YK, Li CC, et al. Genome-wide identification and expression analysis of DXS gene family in Lavandula angustifolia [J]. Genom Appl Biol, 2023, 42(9): 919-926.
[17]	Kelimujiang K, Zhang WY, Zhang XX, et al. Genome-wide investigation of WRKY gene family in Lavandula angustifolia and potential role of LaWRKY57 and LaWRKY75 in the regulation of terpenoid biosynthesis [J]. Front Plant Sci, 2024, 15: 1449299.
[18]	Zhang ZB, Xiong T, Li KJ, et al. The tree of life of copper-containing amine oxidases [J]. Front Plant Sci, 2025, 16: 1544527.
[19]	Jakobsson E, Nilsson J, Ogg D, et al. Structure of human semicarbazide-sensitive amine oxidase/vascular adhesion protein-1 [J]. Acta Crystallogr D Biol Crystallogr, 2005, 61(Pt 11): 1550-1562.
[20]	王鹏, 罗朝鹏, 盖江涛. 茄科植物甲基腐胺氧化酶基因进化路径分析 [J]. 分子植物育种, 2018, 16(1): 47-53.
	Wang P, Luo ZP, Gai JT. Evolutionary path analysis of Methylputrescine oxidase gene in Solanaceae [J]. Mol Plant Breed, 2018, 16(1): 47-53.
[21]	Malli RPN, Adal AM, Sarker LS, et al. De novo sequencing of the Lavandula angustifolia genome reveals highly duplicated and optimized features for essential oil production [J]. Planta, 2019, 249(1): 251-256.
[22]	Qu YN, Wang Q, Guo JH, et al. Peroxisomal CuAOζ and its product H₂O₂ regulate the distribution of auxin and IBA-dependent lateral root development in Arabidopsis [J]. J Exp Bot, 2017, 68(17): 4851-4867.
[23]	Zhang YN, Wang D, Li H, et al. Formation mechanism of glandular trichomes involved in the synthesis and storage of terpenoids in lavender [J]. BMC Plant Biol, 2023, 23(1): 307.
[24]	张夏夏, 陈凌娜, 杨扬, 等. 薰衣草非特异性脂质转移蛋白基因nsLTP2-1和nsLTP2-2的克隆与功能分析 [J]. 植物遗传资源学报, 2024, 25(5): 834-843.
	Zhang XX, Chen LN, Yang Y, et al. Cloning and functional analysis of nonspecific lipid transfer protein genes nsLTP2-1 and nsLTP2-2 in lavender [J]. J Plant Genet Resour, 2024, 25(5): 834-843.
[25]	Meng FL, Zhao HN, Zhu B, et al. Genomic editing of intronic enhancers unveils their role in fine-tuning tissue-specific gene expression in Arabidopsis thaliana [J]. Plant Cell, 2021, 33(6): 1997-2014.
[26]	Wang J, Jiang WL, Yang ZH, et al. Cold-related genes BcCOR15B and BcCOR15A have environment adapted expression pattern in non-heading Chinese cabbage [J]. Plant Physiol Biochem, 2025, 221: 109567.
[27]	秦善, 石洁, 潘徐盈, 等. 植物乳杆菌源胺氧化酶的异源表达及功能结构分析 [J]. 微生物学报, 2023, 63(12): 4698-4713.
	Qin S, Shi J, Pan XY, et al. Heterologous expression, function and structure of amine oxidase derived from Lactiplantibacillus plantarum [J]. Acta Microbiol Sin, 2023, 63(12): 4698-4713.
[28]	倪秀梅, 方芳. 融合表达过氧化氢酶提高多铜氧化酶稳定性及降解生物胺能力 [J]. 生物工程学报, 2021, 37(12): 4382-4394.
	Ni XM, Fang F. Fusion expression with catalase improves the stability of multicopper oxidase and its efficiency in degrading biogenic amines [J]. Chin J Biotechnol, 2021, 37(12): 4382-4394.
[29]	张阿娜, 韩雪, 谷天一, 等. 利用新型红酵母苯丙氨酸解氨酶制备低苯丙氨酸酪蛋白 [J]. 生物技术通报, 2024, 40(8): 309-319.
	Zhang AN, Han X, Gu TY, et al. Preparation of low-phenylalanine casein by novel phenylalanine ammonia-lyases derived from Rhodotorula [J]. Biotechnol Bull, 2024, 40(8): 309-319.
[30]	张婵, 吴友根, 于靖, 等. 光与茉莉酸信号介导的萜类化合物合成分子机制 [J]. 生物技术通报, 2022, 38(8): 32-40.
	Zhang C, Wu YG, Yu J, et al. Molecular mechanism of terpenoids synthesis intermediated by light and jasmonates signals [J]. Biotechnol Bull, 2022, 38(8): 32-40.
[31]	Largeron M. Amine oxidases of the quinoproteins family: their implication in the metabolic oxidation of xenobiotics [J]. Ann Pharm Françaises, 2011, 69(1): 53-61.

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/

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/

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

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

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