High-efficiency secretion expression of FAD-dependent glucose dehydrogenase in Pichia pastoris via signal peptide optimization and redox cofactor enhancement.

Chun Wei, Zhenao Zhang, Jiahao Xu, Jianyong Zheng, Tianhao He, Yuqi Wang, Dongchen Liu, Xueting Chen

Abstract read

In one paragraph

Article in Bioprocess and biosystems engineering, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it

0cells of the map it votes in

0citing 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

0 citing papers in PubMed.

No citing paper in PubMed yet.

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.

Chun WeiKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.

Zhenao ZhangKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.

Jiahao XuKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.

Jianyong ZhengKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. zjy821212@zjut.edu.cn.ORCID https://orcid.org/0000-0002-9821-1062

Tianhao HeKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.

Yuqi WangKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.

Dongchen LiuKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.

Xueting ChenKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.

Funding

Application Development Special Key Project of Chongqing cstc2021jscx-jbgsX0002

6 · The paper itself

Abstract

FAD-dependent glucose dehydrogenase (FAD-GDH) is a promising glucose sensing enzymes owing to its excellent thermostability and substrate specificity. To produce recombinant FAD-GDH with higher enzymatic activity by Pichia pastoris to meet commercial demands, three complementary strategies were proposed. First, the OST signal peptide, which outperformed ADV and MSB, replaced α-MF to yield much more efficient secretion, enhancing enzyme activity and protein yield up to 201.4% and 214.0% compared to the levels obtained with the original secretion signal, respectively, in shake-flask cultures. Second, to address the low specific activity of FAD-GDH, mixed feeding with riboflavin, adenosine, and yeast extract during induction period of fermentation had improved enzyme activity and specific activity to196.6% and 161.4%, respectively, while maintaining cell viability, compared with the control without additional feeding. Third, to further enhance specific activity of FAD-GDH, endogenous FAD synthase was ectopically overexpressed under the control of either the GAP or AOX1 promoter via CRISPR-Cas9-mediated genomic integration. The strain with AOX1-drivenFAD synthase (WCG-AOX1-FAD1) exhibited the most significant improvements, with enzyme activity of 1023.46 ± 95.85 U/mL (239.4% of WCG-OST) and specific activity of 886.31 ± 25.42 U/mg (233.4% of WCG-OST), both representing relatively high levels among similar studies. The synergistic application of these three strategies provides a reference for the industrial production of various cofactor-dependent enzymes.

Indexed as

CRISPR-Cas9Flavin adenine dinucleotideGlucose dehydrogenasePichia pastorisSignal peptide

Identifiers

PMID42862296

DOI10.1002/adsc.201700585

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.