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Characterization of carotenoids in Rhodothermus marinus

机译:海藻红景天中类胡萝卜素的表征

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Rhodothermus marinus, a marine aerobic thermophile, was first isolated from an intertidal hot spring in Iceland. In recent years, the R. marinus strain PRI 493 has been genetically modified, which opens up possibilities for targeted metabolic engineering of the species, such as of the carotenoid biosynthetic pathway. In this study, the carotenoids of the R. marinus type‐strain DSM 4252 T , strain DSM 4253, and strain PRI 493 were characterized. Bioreactor cultivations were used for pressurized liquid extraction and analyzed by ultra‐high performance supercritical fluid chromatography with diode array and quadropole time‐of‐flight mass spectrometry detection (UHPSFC‐DAD‐QTOF/MS). Salinixanthin, a carotenoid originally found in Salinibacter ruber and previously detected in strain DSM 4253, was identified in all three R. marinus strains, both in the hydroxylated and nonhydroxylated form. Furthermore, an additional and structurally distinct carotenoid was detected in the three strains. MS/MS fragmentation implied that the mass difference between salinixanthin and the novel carotenoid structure corresponded to the absence of a 4‐keto group on the ?‐ionone ring. The study confirmed the lack of carotenoids for the strain SB‐71 ( ΔtrpBΔpurAcrtBI’::trpB ) in which genes encoding two enzymes of the proposed pathway are partially deleted. Moreover, antioxidant capacity was detected in extracts of all the examined R. marinus strains and found to be 2–4 times lower for the knock‐out strain SB‐71. A gene cluster with 11 genes in two operons in the R. marinus DSM 4252 T genome was identified and analyzed, in which several genes were matched with carotenoid biosynthetic pathway genes in other organisms.
机译:海洋有氧嗜热菌Rhodothermus marinus最初是从冰岛的潮间带温泉中分离出来的。近年来,已经对R.marinus菌株PRI 493进行了基因修饰,这为该物种的目标代谢工程(例如类胡萝卜素生物合成途径)打开了可能性。在这项研究中,表征了R. marinus型菌株DSM 4252 T,菌株DSM 4253和PRI 493的类胡萝卜素。生物反应器培养物用于加压液体提取,并通过超高性能超临界流体色谱,二极管阵列和四极杆飞行时间质谱检测(UHPSFC-DAD-QTOF / MS)进行分析。 Salinixanthin是一种最初发现于Salinibacter ruber中的类胡萝卜素,先前已在DSM 4253菌株中检测到,在所有三株R. marinus菌株中均被鉴定为羟基和非羟基形式。此外,在这三个菌株中检测到另外的和结构上不同的类胡萝卜素。 MS / MS裂解表明,盐紫质和新的类胡萝卜素结构之间的质量差异对应于β-紫罗兰酮环上不存在4-酮基。这项研究证实了SB-71菌株(ΔtrpBΔpurAcrtBI’:: trpB)中缺乏类胡萝卜素,该菌株中编码拟议途径的两种酶的基因被部分删除。此外,在所有检查过的海藻菌株的提取物中均检测到抗氧化能力,发现其敲除菌株SB-71的抗氧化能力低2-4倍。鉴定并分析了海藻DSM 4252 T基因组中两个操纵子中具有11个基因的基因簇,其中几个基因与其他生物中的类胡萝卜素生物合成途径基因匹配。

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