Capsanthin Production in Escherichia coli by Overexpression of Capsanthin/Capsorubin Synthase from Capsicum annuum
Capsanthin Production in Escherichia coli by Overexpression of Capsanthin/Capsorubin Synthase from Capsicum annuum
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DOI：
10.1021/acs.jafc.1c00083
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发表时间：
2021-04-23
影响因子：
6.1
通讯作者：
Mitani, Yasuo
中科院分区：
文献类型：
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作者：
Furubayashi, Maiko;Kubo, Akiko;Mitani, Yasuo
辣椒素是一种在红辣椒果实中发现的红色类胡萝卜素，作为一种食品和功能性着色添加剂被广泛食用。催化辣椒素合成的酶是capsanthin/capsorubin synthase (CCS)，在20世纪90年代被发现，但没有微生物生产辣椒素的报道。本文报道了利用类胡萝卜素途径工程在大肠杆菌中成功合成辣椒素的首次尝试。我们最初尝试共表达辣椒素生物合成所需的8种酶基因，但没有检测到所需的产物。CCS作为番茄红素β -环化酶和辣椒素/辣椒素合成酶的双重活性可能使这项任务复杂化。我们证明了CCS基因的高表达水平和通过调节7个上游胡萝卜素基因来减少副产物对大肠杆菌中辣椒素的形成至关重要。我们的研究结果为进一步研究微生物中CCS活性和辣椒红素的产生提供了平台。
Capsanthin, a characteristic red carotenoid found in the fruits of red pepper (Capsicum annuum), is widely consumed as a food and a functional coloring additive. An enzyme catalyzing capsanthin synthesis was identified as capsanthin/capsorubin synthase (CCS) in the 1990s, but no microbial production of capsanthin has been reported. We report here the first successful attempt to biosynthesize capsanthin in Escherichia coli by carotenoid-pathway engineering. Our initial attempt to coexpress eight enzyme genes required for capsanthin biosynthesis did not detect the desired product. The dual activity of CCS as a lycopene beta-cyclase as well as a capsanthin/capsorubin synthase likely complicated the task. We demonstrated that a particularly high expression level of the CCS gene and the minimization of byproducts by regulating the seven upstream carotenogenic genes were crucial for capsanthin formation in E. coli. Our results provide a platform for further study of CCS activity and capsanthin production in microorganisms.