Heterologous expression of C30 carotenoid biosynthetic gene crtNM from Lactiplantibacillus plantarum
Heterologous expression of C30 carotenoid biosynthetic gene crtNM from Lactiplantibacillus plantarum
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DOI：
10.1002/jsfa.12160
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发表时间：
2022-08-13
影响因子：
4.1
通讯作者：
Xiong，Zhiqiang
中科院分区：
文献类型：
--
作者：
Gou，Zongqin;Song，Xin;Xiong，Zhiqiang
益生乳酸菌植物乳杆菌（Lactiplantibacillusplantarum）广泛应用于乳制品和其他发酵食品工业。plantarumAR 113含有一个C30类胡萝卜素操纵子crtNM，但C30类胡萝卜素产量仅为8.1 μg g-1DCW。plantarumAR 113基因在大肠杆菌BL 21（DE 3）中得到克隆和重组。基于十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析成功表达了crtN和crtM蛋白，并使用高效液相色谱（HPLC）和液相色谱-质谱（LC-MS）检测了类胡萝卜素。与组成型启动子P44相比，诱导型T7启动子的使用显著提高了E.杆菌优化了发酵条件，用0.5mmol/L IPTG在20 °C诱导7 h。C30类胡萝卜素的产量达到154.5 μg g-1DCW，比L. plantarumAR113.研究了异源表达的C30类胡萝卜素对DPPH和ABTS自由基的清除能力。大肠埃希菌也高于抗氧化食品添加剂二丁基羟基甲苯。大肠杆菌作为乳酸杆菌生产C30类胡萝卜素的基础培养基具有很强的潜力。© 2022化学工业学会。
BACKGROUNDProbiotic lactic acid bacteriumLactiplantibacillus plantarumis widely used in the dairy and other fermented food industries.L. plantarumAR113 harbors a C30carotenoid operoncrtNMbased on genomic analysis, but the yield of C30carotenoid is only 8.1 μg g‐1DCW.RESULTSTo improve the productivity of C30carotenoid,crtNMfromL. plantarumAR113 was cloned and reconstructed inEscherichia coliBL21(DE3). The proteins crtN and crtM were successfully expressed based on a sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS‐PAGE) analysis, and the carotenoid was detected using high‐performance liquid chromatography (HPLC) and liquid chromatography–mass spectrometry (LC–MS). In comparison with the constitutive promoter P44, the use of the inducible T7 promoter significantly increased the carotenoid content inE. coli. The fermentation conditions were also optimized with induction by 0.5 mmol/L IPTG at 20 °C for 7 h. The yield of C30carotenoid reached 154.5 μg g‐1DCW, which was 18‐fold higher than that ofL. plantarumAR113. The 2,2‐diphenyl‐1‐picryl (DPPH) and 2,2′‐azino‐bis (3‐ethylbenzothiazoline‐6sulfonic acid (ABTS) radical scavenging capacity of C30carotenoids synthesized by heterologous expression inE. coliwas also higher than that of the antioxidant food additive butylated hydroxytoluene.CONCLUSIONSOur findings suggest thatE. colihas strong potential as a basic chassis for the production of C30carotenoids fromLactiplantibacilluswith high antioxidant activity. © 2022 Society of Chemical Industry.