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地填写文献信息,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!

Adaptive laboratory evolution accelerated glutarate production by Corynebacterium glutamicum

生物 生化工程 计算生物学 合成生物学 基因 生物合成
作者
Carina Prell,Tobias Busche,Christian Rückert,Lea Nolte,Christoph Brandenbusch,Volker F. Wendisch
出处
期刊:Microbial Cell Factories [BioMed Central]
卷期号:20 (1): 97-97 被引量:3
标识
DOI:10.1186/s12934-021-01586-3
摘要

The demand for biobased polymers is increasing steadily worldwide. Microbial hosts for production of their monomeric precursors such as glutarate are developed. To meet the market demand, production hosts have to be improved constantly with respect to product titers and yields, but also shortening bioprocess duration is important. In this study, adaptive laboratory evolution was used to improve a C. glutamicum strain engineered for production of the C5-dicarboxylic acid glutarate by flux enforcement. Deletion of the l-glutamic acid dehydrogenase gene gdh coupled growth to glutarate production since two transaminases in the glutarate pathway are crucial for nitrogen assimilation. The hypothesis that strains selected for faster glutarate-coupled growth by adaptive laboratory evolution show improved glutarate production was tested. A serial dilution growth experiment allowed isolating faster growing mutants with growth rates increasing from 0.10 h−1 by the parental strain to 0.17 h−1 by the fastest mutant. Indeed, the fastest growing mutant produced glutarate with a twofold higher volumetric productivity of 0.18 g L−1 h−1 than the parental strain. Genome sequencing of the evolved strain revealed candidate mutations for improved production. Reverse genetic engineering revealed that an amino acid exchange in the large subunit of l-glutamic acid-2-oxoglutarate aminotransferase was causal for accelerated glutarate production and its beneficial effect was dependent on flux enforcement due to deletion of gdh. Performance of the evolved mutant was stable at the 2 L bioreactor-scale operated in batch and fed-batch mode in a mineral salts medium and reached a titer of 22.7 g L−1, a yield of 0.23 g g−1 and a volumetric productivity of 0.35 g L−1 h−1. Reactive extraction of glutarate directly from the fermentation broth was optimized leading to yields of 58% and 99% in the reactive extraction and reactive re-extraction step, respectively. The fermentation medium was adapted according to the downstream processing results. Flux enforcement to couple growth to operation of a product biosynthesis pathway provides a basis to select strains growing and producing faster by adaptive laboratory evolution. After identifying candidate mutations by genome sequencing causal mutations can be identified by reverse genetics. As exemplified here for glutarate production by C. glutamicum, this approach allowed deducing rational metabolic engineering strategies.
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
PDF的下载单位、IP信息已删除 (2025-6-4)

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
年年有余完成签到,获得积分10
2秒前
胖小羊完成签到 ,获得积分10
52秒前
1分钟前
领导范儿应助科研通管家采纳,获得10
1分钟前
1分钟前
juan完成签到 ,获得积分10
2分钟前
学术小垃圾完成签到,获得积分10
3分钟前
叁月二完成签到 ,获得积分10
3分钟前
3分钟前
科研通AI2S应助科研通管家采纳,获得10
3分钟前
大模型应助科研通管家采纳,获得10
3分钟前
xingsixs完成签到 ,获得积分10
4分钟前
AprilLeung完成签到 ,获得积分10
5分钟前
5分钟前
深情安青应助科研通管家采纳,获得10
5分钟前
迷茫的一代完成签到,获得积分10
6分钟前
魔笛的云宝完成签到 ,获得积分10
6分钟前
www完成签到,获得积分10
7分钟前
7分钟前
NexusExplorer应助科研通管家采纳,获得10
7分钟前
Akitten发布了新的文献求助10
8分钟前
啥时候吃火锅完成签到 ,获得积分0
9分钟前
上官若男应助科研通管家采纳,获得30
9分钟前
斯文败类应助科研通管家采纳,获得10
9分钟前
李爱国应助科研通管家采纳,获得10
9分钟前
科研通AI2S应助科研通管家采纳,获得10
9分钟前
赘婿应助科研通管家采纳,获得10
9分钟前
田様应助精明晓刚采纳,获得10
10分钟前
10分钟前
精明晓刚发布了新的文献求助10
10分钟前
科研通AI2S应助科研通管家采纳,获得10
11分钟前
coolplex完成签到 ,获得积分10
12分钟前
wwe完成签到,获得积分10
12分钟前
貔貅完成签到 ,获得积分10
13分钟前
yindi1991完成签到 ,获得积分10
15分钟前
hgl完成签到,获得积分10
15分钟前
15分钟前
科研通AI2S应助科研通管家采纳,获得10
15分钟前
15分钟前
华仔应助精明晓刚采纳,获得10
16分钟前
高分求助中
The Mother of All Tableaux: Order, Equivalence, and Geometry in the Large-scale Structure of Optimality Theory 3000
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 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 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3990423
求助须知:如何正确求助?哪些是违规求助? 3532158
关于积分的说明 11256513
捐赠科研通 3271046
什么是DOI,文献DOI怎么找? 1805207
邀请新用户注册赠送积分活动 882302
科研通“疑难数据库(出版商)”最低求助积分说明 809234