Development of an efficient autoinducible expression system by promoter engineering in Bacillus subtilis

枯草芽孢杆菌 绿色荧光蛋白 质粒 生物 突变体 异源的 发起人 分子生物学 诱导剂 基因表达 生物化学 化学 基因 细菌 遗传学
作者
Chengran Guan,Wenjing Cui,Jintao Cheng,Li Zhou,Zhongmei Liu,Zhemin Zhou
出处
期刊:Microbial Cell Factories [BioMed Central]
卷期号:15 (1) 被引量:70
标识
DOI:10.1186/s12934-016-0464-0
摘要

Bacillus subtilis, a Gram-positive organism, has been developed to be an attractive expression platform to produce both secreted and cytoplasmic proteins owing to its prominent biological characteristics. We previously developed an auto-inducible expression system containing the srfA promoter (PsrfA) which was activated by the signal molecules acting in the quorum-sensing pathway for competence. The P srfA promoter exhibited the unique property of inducer-free activity that is closely correlated with cell density.To improve the PsrfA-mediated expression system to the high-cell-density fermentation for industrial production in the B. subtilis mutant strain that is unable to sporulate, a spore mutant strain BSG1682 was developed, and the PsrfA promoter was enhanced by promoter engineering. Using green fluorescent protein (GFP) as the reporter, higher fluorescent intensity was observed in BSG1682 with expression from either plasmid or chromosome than that of the wild type B. subtilis 168. Thereafter, the PsrfA was engineered, yielding a library of PsrfA derivatives varied in the strength of GFP expression. The P23 promoter exhibited the best performance, almost twofold stronger than that of P srfA. Two heterologous proteins, aminopeptidase (AP) and nattokinase (NK), were successfully overproduced under the control of P23 in BSG1682. Finally, the capacity of the expression system was demonstrated in batch fermentation in a 5-L fermenter.The expression system demonstrates prominence in the activity of the auto-inducible promoter. Desired proteins could be highly and stably produced by integrating the corresponding genes downstream of the promoter on the plasmid or the chromosome in strain BSG1682. The expression system is conducive to the industrial production of pharmaceuticals and heterologous proteins in high-cell-density fermentation in BSG1682.
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
刚刚
baishui发布了新的文献求助10
刚刚
为神指路发布了新的文献求助10
刚刚
1秒前
FashionBoy应助咿咿呀呀采纳,获得10
2秒前
2秒前
淡淡绮琴发布了新的文献求助10
3秒前
3秒前
猴子大王666完成签到,获得积分10
3秒前
orixero应助future采纳,获得10
3秒前
勤劳茗发布了新的文献求助10
4秒前
追寻的怜容完成签到,获得积分10
4秒前
5秒前
一一高速下载文献没有问题完成签到,获得积分10
6秒前
熙欢完成签到,获得积分10
6秒前
白藏发布了新的文献求助10
6秒前
1351567822应助ake采纳,获得10
7秒前
Holly12345应助xiao采纳,获得10
7秒前
陈阳发布了新的文献求助10
7秒前
xiao双月完成签到,获得积分10
8秒前
随风走发布了新的文献求助10
8秒前
9秒前
不爱吃姜完成签到,获得积分10
9秒前
Rocc完成签到,获得积分10
10秒前
华仔应助美满花生采纳,获得30
10秒前
沈竑宇完成签到,获得积分10
10秒前
coco完成签到,获得积分10
11秒前
深情安青应助王者归来采纳,获得30
11秒前
李健的小迷弟应助shinn采纳,获得10
11秒前
咿咿呀呀发布了新的文献求助10
12秒前
12秒前
上官若男应助猴子大王666采纳,获得10
12秒前
李冠龙应助石会发采纳,获得10
13秒前
不爱吃姜发布了新的文献求助10
14秒前
难过梦竹发布了新的文献求助40
15秒前
youchgg完成签到,获得积分10
15秒前
future发布了新的文献求助10
15秒前
呼安完成签到,获得积分10
16秒前
16秒前
Asteria发布了新的文献求助10
17秒前
高分求助中
Picture Books with Same-sex Parented Families: Unintentional Censorship 700
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
Nucleophilic substitution in azasydnone-modified dinitroanisoles 500
不知道标题是什么 500
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
Effective Learning and Mental Wellbeing 300
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 3975339
求助须知:如何正确求助?哪些是违规求助? 3519670
关于积分的说明 11199199
捐赠科研通 3256002
什么是DOI,文献DOI怎么找? 1798043
邀请新用户注册赠送积分活动 877386
科研通“疑难数据库(出版商)”最低求助积分说明 806305