亲爱的研友该休息了!由于当前在线用户较少,发布求助请尽量完整的填写文献信息,科研通机器人24小时在线,伴您度过漫漫科研夜!身体可是革命的本钱,早点休息,好梦!

Production of Carminic Acid by Metabolically Engineered Escherichia coli

化学 大肠杆菌 发酵 代谢工程 发光光杆线虫 葡萄糖基转移酶 羟基化 聚酮 生物化学 生物合成 基因
作者
Dongsoo Yang,Woo Dae Jang,Sang Yup Lee
出处
期刊:Journal of the American Chemical Society [American Chemical Society]
卷期号:143 (14): 5364-5377 被引量:35
标识
DOI:10.1021/jacs.0c12406
摘要

Carminic acid is an aromatic polyketide found in scale insects (i.e., Dactylopius coccus) and is a widely used natural red colorant. It has long been produced by the cumbersome farming of insects followed by multistep purification processes. Thus, there has been much interest in producing carminic acid by the fermentation of engineered bacteria. Here we report the complete biosynthesis of carminic acid from glucose in engineered Escherichia coli. We first optimized the type II polyketide synthase machinery from Photorhabdus luminescens, enabling a high-level production of flavokermesic acid upon coexpression of the cyclases ZhuI and ZhuJ from Streptomyces sp. R1128. To discover the enzymes responsible for the remaining two reactions (hydroxylation and C-glucosylation), biochemical reaction analyses were performed by testing enzyme candidates reported to perform similar reactions. The two identified enzymes, aklavinone 12-hydroxylase (DnrF) from Streptomyces peucetius and C-glucosyltransferase (GtCGT) from Gentiana triflora, could successfully perform hydroxylation and C-glucosylation of flavokermesic acid, respectively. Then, homology modeling and docking simulations were performed to enhance the activities of these two enzymes, leading to the generation of beneficial mutants with 2–5-fold enhanced conversion efficiencies. In addition, the GtCGT mutant was found to be a generally applicable C-glucosyltransferase in E. coli, as was showcased by the successful production of aloesin found in Aloe vera. Simple metabolic engineering followed by fed-batch fermentation resulted in 0.63 ± 0.02 mg/L of carminic acid production from glucose. The strategies described here will be useful for the design and construction of biosynthetic pathways involving unknown enzymes and consequently the production of diverse industrially important natural products.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
3秒前
xyw发布了新的文献求助10
8秒前
上官若男应助chh采纳,获得10
10秒前
12秒前
12秒前
15秒前
头大四年发布了新的文献求助10
17秒前
18秒前
sissiarno应助科研通管家采纳,获得30
21秒前
21秒前
sissiarno应助科研通管家采纳,获得30
21秒前
慕青应助高兴的无颜采纳,获得10
21秒前
22秒前
25秒前
饭团不吃鱼完成签到,获得积分10
25秒前
25秒前
慕青应助走走采纳,获得10
27秒前
chh发布了新的文献求助10
27秒前
西早发布了新的文献求助10
28秒前
狼牧羊城完成签到,获得积分10
39秒前
高兴的无颜完成签到,获得积分10
42秒前
pegasus0802完成签到,获得积分10
44秒前
飘着的鬼完成签到,获得积分10
45秒前
49秒前
51秒前
55秒前
走走发布了新的文献求助10
55秒前
1分钟前
GT完成签到,获得积分10
1分钟前
体育爱好者完成签到,获得积分10
1分钟前
旺旺小小贝完成签到,获得积分10
1分钟前
牟白容发布了新的文献求助30
1分钟前
牟白容完成签到,获得积分10
1分钟前
1分钟前
落后的忆文关注了科研通微信公众号
1分钟前
1分钟前
1分钟前
hillson发布了新的文献求助10
1分钟前
头大四年发布了新的文献求助10
1分钟前
hank完成签到,获得积分10
1分钟前
高分求助中
Sustainability in Tides Chemistry 2000
Microlepidoptera Palaearctica, Volumes 1 and 3 - 13 (12-Volume Set) [German] 1122
Diamonds: Properties, Synthesis and Applications 1000
The Data Economy: Tools and Applications 1000
Mantiden - Faszinierende Lauerjäger – Buch gebraucht kaufen 700
PraxisRatgeber Mantiden., faszinierende Lauerjäger. – Buch gebraucht kaufe 700
A Dissection Guide & Atlas to the Rabbit 600
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3099715
求助须知:如何正确求助?哪些是违规求助? 2751149
关于积分的说明 7611895
捐赠科研通 2402905
什么是DOI,文献DOI怎么找? 1275047
科研通“疑难数据库(出版商)”最低求助积分说明 616226
版权声明 599033