Evidence map›Paper›PMID 42101543›Full record

ArticleWorld journal of microbiology & biotechnology2026

rDNA-mediated multicopy integration and gene dosage quantification system for microbial zeaxanthin biosynthesis.

Ruijing Ling, Shuting Hou, Xiangzhao Mao, Francesco Secundo, Wei Zhou, Meiyi Xi, Yang Xiao, Bei Gao

Abstract read
PubMed Publisher
In one paragraph

Article in World journal of microbiology & biotechnology, 2026. 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
–field-weighted citation impact
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.

  1. Transposon engineering ofEngineering in life sciences · 2026
    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

8 authors.

Ruijing LingState Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, P.O.B.311, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
Shuting HouState Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, P.O.B.311, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
Xiangzhao MaoCollege of Food Science and Engineering, State Key Laboratory of Marine Food Processing and Safety Control, Ocean University of China, Qingdao, Shandong, 266404, People's Republic of China.
Francesco SecundoNational Research Council, Giulio Natta Institute of Chemical Sciences and Technologies, Via Mario Bianco 9, Milan, I-20131, Italy.
Wei ZhouState Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, P.O.B.311, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
Meiyi XiState Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, P.O.B.311, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
Yang XiaoState Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, P.O.B.311, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
Bei GaoState Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, P.O.B.311, 130 Meilong Road, Shanghai, 200237, People's Republic of China. gaobei@ecust.edu.cn.

Funding

National Key Research and Development Program of China No. 2024YFA0917900Shanghai Science and Technology Innovation Action Plan grant no. 23HC1400500
6 · The paper itself

Abstract

Zeaxanthin is an oxygenated carotenoid with established physiological functions in ocular health and antioxidant protection. Currently, industrial zeaxanthin production relies primarily on plant extraction and chemical synthesis, which is costly and poses a significant environmental burden. Microbial biosynthesis offers a sustainable and eco-friendly alternative, where the gene dosage of key enzymes critically influences biosynthetic performance. However, optimizing copy number remains challenging for pathways requiring tandem insertion of introduced genes into limited genomic loci. Accordingly, this study developed a genetic system rNTS, which targets the ribosomal non-transcribed spacer region for one-step, high-copy integration of multiple genes and direct assessment of gene dosage effects. Fluorescence Intensity Ratios (FIRs) served as indicators for the direct and quantitative estimation of gene copy number and the selection of optimal dosage. An individual rNTSi (integration) vector integrated up to 40 single-gene copies, while parallel rNTSi vectors enabled multi-gene strains totaling 20 copies with independent visual selection. Following marker removal with rNTSr (removal) vector via the Cre/LoxP system, selection markers were efficiently reused for iterative integrations to achieve 32 copies. Applying rNTS to zeaxanthin biosynthesis rapidly identified the optimal dosage of the rate-limiting enzyme CrtZ, yielding titers of 1.10 g/L under glucose fermentation and 1.19 g/L under methanol-induced fermentation. Owing to its high efficiency, multi-gene copy number readability, and iterative integration capacity, the rNTS strategy offers an additional solution for quantitative gene-dosage assessment and microbial biosynthetic pathway optimization.

Indexed as

DNA, RibosomalGene DosageZeaxanthinsBiosynthetic PathwaysGenetic VectorsMetabolic EngineeringDNA, RibosomalZeaxanthinsCopy number readabilityIterative integrationKomagataella phaffiirDNAZeaxanthin

Identifiers

PMID42101543

What OpenQuestion holds

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Registered trials

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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.