Evidence map›Paper›PMID 33405008›Full record

ArticleApplied biochemistry and biotechnology2021

Optimization of Cephalosporin C Acylase Expression in Escherichia coli by High-Throughput Screening a Constitutive Promoter Mutant library.

Hongxu Sun, Tianjiao Liu, Hui Luo, Zihao Nie, Yanhong Chang, Huimin Yu, Zhongyao Shen

Abstract read
PubMed Publisher
In one paragraph

Article in Applied biochemistry and biotechnology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
0.1field-weighted citation impact, top 59% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed, 6 citations in OpenAlex.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors at 2 institutions in 1 country.

Hongxu Sun *Department of Biological Science and Engineering, University of Science and Technology Beijing, Beijing, 100083, China.
Tianjiao Liu *Department of Biological Science and Engineering, University of Science and Technology Beijing, Beijing, 100083, China.
Hui LuoDepartment of Biological Science and Engineering, University of Science and Technology Beijing, Beijing, 100083, China. luohui@ustb.edu.cn.ORCID http://orcid.org/0000-0002-3075-2129
Zihao NieDepartment of Biological Science and Engineering, University of Science and Technology Beijing, Beijing, 100083, China.
Yanhong ChangDepartment of Environmental Engineering, University of Science and Technology Beijing, Beijing, 100083, China.
Huimin YuDepartment of Chemical Engineering, Tsinghua University, Beijing, 100084, China.
Zhongyao ShenDepartment of Chemical Engineering, Tsinghua University, Beijing, 100084, China.
University of Science and Technology Beijing · CNTsinghua University · CN

Funding

National Natural Science Foundation of China 21276023National Natural Science Foundation of China 21476025
6 · The paper itself

Abstract

Cephalosporin C acylase (CCA) is capable of catalyzing cephalosporin C (CPC) to produce 7-aminocephalosporanic acid (7-ACA), an intermediate of semi-synthetic cephalosporins. Inducible expression is usually used for CCA. To improve the efficiency of CCA expression without gene induction, three recombinant strains regulated by constitutive promoters BBa_J23105, PLtetO1, and tac were constructed, respectively. Among them, BBa_J23105 was the best promoter and its mutant libraries were established using saturation mutagenesis. In order to obtain the mutants with enhanced activity, a high-throughput screening method based on flow cytometric sorting techniques was developed by using green fluorescent protein (GFP) as the reporter gene. A series of mutants were screened at 28 °C, 200 rpm, and 24-h culture condition. The study of mutants showed that the enzyme activity, fluorescence intensity, and promoter transcriptional strength were positively correlated. The enzyme activity of the optimal mutant obtained by screening reached 12772 U/L, 3.47 times that of the original strain.

Indexed as

Escherichia coliEscherichia coli ProteinsGene Expression Regulation, BacterialGene Expression Regulation, EnzymologicGene LibraryMutationPenicillin AmidasePromoter Regions, GeneticEscherichia coli ProteinsPenicillin AmidaseCephalosporin C acylaseConstitutive promoterFlow cytometric screeningGreen fluorescent proteinPromoter engineering

Identifiers

PMID33405008
OpenAlexW3120595257

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.