Molecular and Enzymatic Characterization of 9-Cis-epoxycarotenoid Dioxygenases from Mulberry

IF 1.9 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

The Protein Journal Pub Date : 2022-08-13 DOI:10.1007/s10930-022-10072-7

Dan Liu, Changyu Qiu, Yanrong Zeng, Qiang Lin

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Abstract

Abscisic acid (ABA) is involved in many physiological regulatory processes in plants, such as leaf shedding, stomatal closure, inhibition of cell elongation, as well as responses to multi-abiotic stress, and 9-cis epoxy carotenoid dioxygenase (NCED) is related to the indirect synthesis of ABA. However, NCED genes involved in multi-abiotic stress and ABA synthesis pathway in mulberry (Morus alba L.) are still unknown. Here, two NCED genes cloned from mulberry (MaNCED) and their function were preliminarily identified. Interestingly, MaNCED2 responded strongly to drought stress while MaNCED1 responded strongly to pathogen stress. Then, two MaNCED proteins were successfully obtained by prokaryotic expression, and the degradation products of MaNCED1 and MaNCED2 were analyzed using UPLC-MS. The results show that recombinant MaNCED1 and MaNCED2 both cleave 9-cis-violaxanthin to form C₁₅ xanthoxin, involved in the formation of the precursor of ABA.

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桑树9-顺式环氧类胡萝卜素双加氧酶的分子和酶学表征

脱落酸(ABA)参与植物叶片脱落、气孔关闭、抑制细胞伸长以及对多生物胁迫的响应等生理调控过程，而9-顺式环氧类胡萝卜素双加氧酶(NCED)与ABA的间接合成有关。然而，桑树(Morus alba L.)中参与多生物胁迫和ABA合成途径的NCED基因尚不清楚。本文从桑树中克隆了两个NCED基因(MaNCED)，并对其功能进行了初步鉴定。有趣的是，MaNCED2对干旱胁迫反应强烈，而MaNCED1对病原体胁迫反应强烈。然后，通过原核表达成功获得了两个MaNCED蛋白，并使用UPLC-MS分析了MaNCED1和MaNCED2的降解产物。结果表明，重组蛋白MaNCED1和MaNCED2均可裂解9-顺式紫黄质形成C15黄质，参与ABA前体的形成。

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来源期刊

The Protein Journal 生物-生化与分子生物学

CiteScore

5.20

自引率

0.00%

发文量

审稿时长

12 months

期刊介绍： The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.