High-content screening of Aspergillus niger with both increased production and high secretion rate of glucose oxidase

Zhu, Xu-Dong; Sun, Jingchun; Chu, Ju

doi:10.1007/s10529-017-2442-y

Cited by 4 publications

(4 citation statements)

References 16 publications

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“…b The differences between the yield of SLs and glycerol consumption of mutants and wild-type strain were taken as x-axis and y-axis, respectively Compared with single index, two indexes can improve efficiency of screening for high-performance strains. Zhu et al (2018) developed a rapid, dual-parameter, platebased screening process to simultaneously improve the production and secretion rate of glucose oxidase in Aspergillus niger. Using this dual-parameter screening method, a strain with glucose oxidase activity 164% higher than the original strain was obtained.…”

Section: Discussionmentioning

confidence: 99%

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Rational high-throughput screening system for high sophorolipids production in Candida bombicola by co-utilizing glycerol and glucose capacity

Lin

Yang

et al. 2019

Bioresour. Bioprocess.

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Background: A rational and high-performance high-throughput screening system effectively improves the efficiency of strain screening. Results: A rapid and efficient method was developed to detect carbon sources. And then by using glucose analog as screening pressure and two indexes for sophorolipids (SLs) production and glycerol consumption, a mutant of Candida bombicola L1.3 was successfully selected among 3000 mutants. It produces high SLs titer and co-utilizes glucose and glycerol simultaneously. Conclusions: Compared to the wild-type strain, L1.3 exhibited 15.06% higher SLs titer and 35.69% higher glycerol consumption capacity in a 5-L bioreactor. We believe that L1.3 can potentially be used for the efficient industrial production of SLs with simple downstream processing for separating SLs and glycerol.

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Section: Discussionmentioning

confidence: 99%

“…In general, glucose analogs such as 2-deoxy-d-glucose (Afanas'Eva and Burd 1980) were used for breeding strains with eliminated or alleviated carbon catabolic repression (Zhu et al 2018). On the other hand, a highperformance producing strain can increase the competitiveness of the companies.…”

Section: Introductionmentioning

confidence: 99%

Rational high-throughput screening system for high sophorolipids production in Candida bombicola by co-utilizing glycerol and glucose capacity

Lin

Yang

et al. 2019

Bioresour. Bioprocess.

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“…Aspergillus niger is a member of the filamentous fungi [1], which is widely used for the production of enzymes [2–4] and organic acids [5–7]. In the era of functional genomics, the in‐depth study of A. niger at the molecular genetic level could promote basic research around it and expand its usage in industry, agriculture, and other domains.…”

Section: Introductionmentioning

confidence: 99%

In vitro CRISPR/Cas9 system for genome editing of Aspergillus niger based on removable bidirectional selection marker AmdS

Nan

Ouyang

Chu

2020

Biotech and App Biochem

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We established an in vitro clustered regularly interspaced short palindromic repeats (CRISPR)‐associated RNA‐guided DNA endonucleases (Cas9) system to efficiently produce specific genome editing in Aspergillus niger, using a novel recyclable, bidirectional selection marker gene amdS without the need of prior production of an amdS mutant. The donor DNA plasmid consisted of amdS open reading frame, promoter, terminator, and directional repeats (DRs) flanking sequences. It was cotransformed with recombinant nuclease Cas9 and the sgRNA, which targets to the pigment gene olvA of A. niger strain CBS513.88. The positive olive transformants, other than the wild‐type strain, were able to grow on the media containing acetamide as the sole nitrogen source and cesium chloride. Furthermore, culturing the transformants on media with fluoroacetamide and urea allowed a loop‐out of the amdS expression cassette by recombining the flanking DRs. This study confirmed the facts that the endogenous amdS can be used as a dominant marker and that it can be removed by counter‐selection in gene editing of A. niger. The proposed in vitro CRISPR/Cas9 method offers a powerful tool for marker‐free genetic manipulation of filamentous fungi industrial‐specific strains.

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“…Aspergillus niger, as a member of filamentous fungi, is widely used to produce and secrete a variety of bioactive substance including organic acids, like citric acid (Wang et al 2017), oxalic acid (Lee et al 2018), gallic acid (Mata-Gomez et al 2015) as well as many carbohydrate-active enzyme (CAzymes), such as glucoamylase (Suyama et al 2017), glucose oxidase (Zhu et al 2018), and amylase (Varalakshmi et al 2009), which are valuable in industry especially food industry. And it is generally recognized as safe (GRAS) by the United States Food and Drug Administration.…”

Section: Introductionmentioning

confidence: 99%

Efficient gene deletion and replacement in Aspergillus niger by modified in vivo CRISPR/Cas9 systems

Zhang

Ouyang

Nan

et al. 2019

Bioresour. Bioprocess.

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Aspergillus niger, as an important industrial strain, is widely used in the production of a variety of organic acids and industrial enzymes. To excavate the greater potential of A. niger as a cell factory, the development of highly efficient genome editing techniques is crucial. Here, we developed a modified CRISPR/Cas9 system for A. niger highlighted in two aspects: (1) construction of a single and easy-to-use CRISPR/Cas9 tool plasmid derived from pAN7-1 which is widely used in filamentous fungi; (2) redesign of the easy-to-switch "ribozyme-gRNA-ribozyme (RGR)" element in the tool plasmid. We examined the gene inactivation efficiency without repair fragment and the gene replacement efficiency with repair fragment utilizing the modified system, respectively, and both of them reach the efficiency as high as over 90%. Especially, the co-transformation of the tool plasmid and the specific repair fragment can easily realize one-step knock-out/knock-in of target genes, even with the length of homologous arms as only 100 bp. The establishment of this system will lay a solid foundation for the gene function research and rational design of cell factory in A. niger or broader filamentous fungi hosts. which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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High-content screening of Aspergillus niger with both increased production and high secretion rate of glucose oxidase

Abstract: Using this dual-parameter screening method, we acquired a strain with GOD activity of 3126 U l, which was 146% higher than the original strain. Its secretion rate of GOD was 83, 32% higher than the original strain.

Cited by 4 publications

References 16 publications

Rational high-throughput screening system for high sophorolipids production in Candida bombicola by co-utilizing glycerol and glucose capacity

Rational high-throughput screening system for high sophorolipids production in Candida bombicola by co-utilizing glycerol and glucose capacity

In vitro CRISPR/Cas9 system for genome editing of Aspergillus niger based on removable bidirectional selection marker AmdS

Efficient gene deletion and replacement in Aspergillus niger by modified in vivo CRISPR/Cas9 systems

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