植物乳植杆菌类胡萝卜素生物合成关键基因LpispA的功能鉴定

Functional Characterization of LpispA,a Key Gene Involved in Carotenoid Biosynthesis in Lactiplantibacillus plantarum

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中文摘要：类胡萝卜素具有抗氧化和抗肿瘤等多种重要生理活性.与化学合成法和植物提取法相比较,利用微生物法合成新型的、高价值的类胡萝卜素,具有食用安全和经济可行等优势.植物乳植杆菌作为天然C30 类胡萝卜素来源的几种细菌之一,其类胡萝卜素生物合成途径却尚未被阐明清楚.该论文基于转录组数据对一株海洋源植物乳植杆菌中的LpispA基因进行克隆、绘制系统进化树,并构建了类胡萝卜素合成功能互补的大肠杆菌表达系统pAC-BETA∆E-ispA,再采用紫外可见光谱法和反相高效液相色谱法验证产物成分.结果表明,植物乳植杆菌LpispA与桃色欧文氏菌的亲缘关系最近;构建的大肠杆菌工程菌株pAC-BETA∆E-ispA的β-胡萝卜素(C40)产量为0.05 mg/g DCW.因此,LpispA在大肠杆菌功能互补表达系统中成功表达了Geranylgeranyl Pyrophosphate(GGPP)合成酶的活性,也即说明LpispA在细菌类胡萝卜素合成途径中可作为类胡萝卜素合成多功能酶,共同参与C30 和C40 类胡萝卜素的合成.该研究为植物乳植杆菌类胡萝卜素生物合成机制研究和利用植物乳植杆菌作为类胡萝卜素补充剂的应用奠定了实验基础.

外文摘要：Carotenoids exhibit various important physiological activities,including anti-oxidation and anti-tumor properties.Compared with plant extracts and chemically synthesized compounds,carotenoids are considered safe for human consumption.Moreover,microbial biosynthesis offers an economically viable approach to the synthesis of carotenoids.As a novel source of the carotenoid C30,the carotenoid biosynthetic pathway in Lactiplantibacillus plantarum has not been fully elucidated.Based on transcriptome analyses performed previously,in this study,the LpispA gene was cloned from a marine L.plantarum strain,and phylogenetic analysis was performed.Subsequently,the cloned open reading frame sequence was expressed in the reconstructed plasmid pAC-BETA∆E-ispA within a functional complementation system using Escherichia coli for carotenoid biosynthesis.In addition,ultraviolet-visible light spectroscopy and reversed-phase high-performance liquid chromatography were employed for component detection of the pigment productions.The results indicated that the L.plantarum LpispA gene exhibited the closest genetic relationship with the corresponding gene from Erwinia persicina.A β-carotene(C40)yield of 0.05 mg/g(dry cell weight)was achieved in the engineered E.coli strain with pAC-BETA∆E-ispA.In conclusion,the bioactivity of geranylgeranyl pyrophosphate synthase expressed by LpispA in the functional complementation E.coli system resulted in the biosynthesis of β-carotene.In other words,LpispA,isolated from L.plantarum,is a multifunctional enzyme in carotenoid biosynthesis,contributing to the biosynthesis of both C30 and C40 carotenoids in bacteria.These results may establish an experimental foundation for investigating carotenoid biosynthesis in L.plantarum and pave the way for potential applications of L.plantarum in the production of carotenoid supplements.

外文关键词：

carotenoidsLactiplantibacillus plantarumLpispA genecomplementary expression

作者：

李舒丽、谢卓婷、沈润泽、郭丽琼、林俊芳、陈谋通、叶志伟

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作者单位：

华南农业大学食品学院,广东广州 510642

广东省微生态制剂工程技术研究中心,广东广州 510642

广东省科学院微生物研究所,华南应用微生物国家重点实验室,广东省微生物安全与健康重点实验室,农业农村部农业微生物组学与精准应用重点实验室,广东广州 510070

关键词：

类胡萝卜素植物乳植杆菌 LpispA基因互补表达

基金：

国家自然科学基金项目国家自然科学基金项目广东省自然科学基金项目广东省自然科学基金项目广东省重点领域研发计划重点项目广东省农业科研项目和农业技术推广项目广东省农业科研项目和农业技术推广项目

项目编号：

32272298318019182022A15150100572015A0303102252022B02020500022022KJ1032023KJ103

出版年：

2024

DOI：

10.13982/j.mfst.1673-9078.2024.2.0215

现代食品科技

华南理工大学

现代食品科技

CSTPCD北大核心

影响因子：1.07

ISSN：1673-9078

年,卷(期)：2024.40(2)

参考文献量29