Characterization of a flavin-containing monooxygenase from Corynebacterium glutamicum and its application to production of indigo and indirubin
Characterization of a flavin-containing monooxygenase from Corynebacterium glutamicum and its application to production of indigo and indirubin
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DOI：
10.1007/s10529-015-1824-2
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发表时间：
2015-08-01
影响因子：
2.7
通讯作者：
Lee, Jin Ho
中科院分区：
文献类型：
--
作者：
Ameria, Sisi Patricia Lolita;Jung, Hye Sook;Lee, Jin Ho
目的 研究谷氨酸棒杆菌 ATCC13032 中编码黄素单加氧酶 (cFMO) 的基因在大肠杆菌中克隆和表达时在靛蓝色素生产中的作用。重组大肠杆菌产生的蓝色色素被鉴定为靛蓝和靛玉红。 cFMO 被纯化为与麦芽糖结合蛋白 (MBP) 的融合形式。该酶在 25 A 摄氏度和 pH 8 时最佳。根据吸收光谱分析，cFMO 被归类为黄素蛋白。 FMO 活性被 1 mM Cu2+ 强烈抑制，并通过添加 1-10 mM EDTA 恢复。该酶催化TMA、硫脲和半胱胺的氧化，但不催化谷胱甘肽或半胱氨酸的氧化。 MBP-cFMO 通过将吲哚氧化成吲哚酚而具有吲哚加氧酶活性。重组大肠杆菌从2.5 g l-色氨酸l(-1) 中产生685 mg 靛蓝l(-1) 和103 mg 靛红l(-1)。结果表明cFMO 可用于微生物生产靛蓝和靛玉红。
To examine the role of a gene encoding flavin-containing monooxygenase (cFMO) from Corynebacterium glutamicum ATCC13032 when cloned and expressed in Escherichia coli for the production of indigo pigments.The blue pigments produced by recombinant E. coli were identified as indigo and indirubin. The cFMO was purified as a fused form with maltose-binding protein (MBP). The enzyme was optimal at 25 A degrees C and pH 8. From absorption spectrum analysis, the cFMO was classified as a flavoprotein. FMO activity was strongly inhibited by 1 mM Cu2+ and recovered by adding 1-10 mM EDTA. The enzyme catalyzed the oxidation of TMA, thiourea, and cysteamine, but not glutathione or cysteine. MBP-cFMO had an indole oxygenase activity through oxygenation of indole to indoxyl. The recombinant E. coli produced 685 mg indigo l(-1) and 103 mg indirubin l(-1) from 2.5 g l-tryptophan l(-1).The results suggest the cFMO can be used for the microbial production of both indigo and indirubin.