A high-throughput method for screening of Aspergillus niger mutants with high transglycosylation activity by detecting non-fermentable reducing sugar

Chen, Guiguang; Li, Wei; Zhang, Yun-Kai; Qin, Yongling; Wu, Kong-Yang; Liang, Zhiqun

doi:10.1007/s11274-010-0595-0

Cited by 10 publications

(3 citation statements)

References 19 publications

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“…Escherichia coli DH5α, Lactobacillus sp. PL17, Pichia pastoris GS115, Saccharomyces cerevisiae GXJ-1 (Chen et al, 2011), and Aspergillus niger GS3-3 (Li et al, 2014) preserved in our lab were used as indicator strains for antimicrobial assay.…”

Section: Methodsmentioning

confidence: 99%

Non-sterile Submerged Fermentation of Fibrinolytic Enzyme by Marine Bacillus subtilis Harboring Antibacterial Activity With Starvation Strategy

Pan

Chen

et al. 2019

Front. Microbiol.

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Microbial fibrinolytic enzyme is a promising candidate for thrombolytic therapy. Non-sterile production of fibrinolytic enzyme by marine Bacillus subtilis D21-8 under submerged fermentation was realized at a mild temperature of 34°C, using a unique combination of starvation strategy and self-production of antibacterial agents. A medium composed of 18.5 g/L glucose, 6.3 g/L yeast extract, 7.9 g/L tryptone, and 5 g/L NaCl was achieved by conventional and statistical methods. Results showed efficient synthesis of fibrinolytic enzyme and antibacterial compounds required the presence of both yeast extract and tryptone in the medium. At shake-flask level, the non-sterile optimized medium resulted in higher productivity of fibrinolytic enzyme than the sterile one, with an enhanced yield of 3,129 U/mL and a production cost reduced by 24%. This is the first report dealing with non-sterile submerged fermentation of fibrinolytic enzyme, which may facilitate the development of feasible techniques for non-sterile production of raw materials for the preparation of potential drugs with low operation cost.

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Section: Methodsmentioning

confidence: 99%

Non-sterile Submerged Fermentation of Fibrinolytic Enzyme by Marine Bacillus subtilis Harboring Antibacterial Activity With Starvation Strategy

Pan

Chen

et al. 2019

Front. Microbiol.

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“…Aspergillus sp. ( A. niger , A. oryzae , A. terreus , A. flavus ) possess natural advantages in the production of enzymes, organic acids, and other high-value products due to their powerful hydrolytic enzyme system and strong protein secretion pathway that enable it to grow and reproduce quickly and adapt to harsh environments [ 77 – 80 ]. Furthermore, researchers developed strategies for more efficient production based on high protein secretion efficiency, genomic information, and low culture costs[ 4 ].…”

Section: Applications Of Filamentous Fungi As Microbial Cell Factoriesmentioning

confidence: 99%

“…Strategies for mutant screening mainly include high-throughput breeding, morphology screening, and adaptive laboratory evolution, which is the direct method to increase the efficiency of microbial cell factories [ 4 , 81 ]. High-throughput breeding was successfully applied to enhance antifungal activity, glucosidase activity, and gluconate production by improving cellular metabolism [ 78 , 80 , 82 ]. For example, resazurin as used as the indicator for high-throughput screening to improve antifungal activity, which was successfully identified in 2.7% of 12,000 microbial extracts [ 83 ].…”

Section: Applications Of Filamentous Fungi As Microbial Cell Factoriesmentioning

confidence: 99%

Microbial cell factories based on filamentous bacteria, yeasts, and fungi

Ding

2023

Microb Cell Fact

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Background Advanced DNA synthesis, biosensor assembly, and genetic circuit development in synthetic biology and metabolic engineering have reinforced the application of filamentous bacteria, yeasts, and fungi as promising chassis cells for chemical production, but their industrial application remains a major challenge that needs to be solved. Results As important chassis strains, filamentous microorganisms can synthesize important enzymes, chemicals, and niche pharmaceutical products through microbial fermentation. With the aid of metabolic engineering and synthetic biology, filamentous bacteria, yeasts, and fungi can be developed into efficient microbial cell factories through genome engineering, pathway engineering, tolerance engineering, and microbial engineering. Mutant screening and metabolic engineering can be used in filamentous bacteria, filamentous yeasts (Candida glabrata, Candida utilis), and filamentous fungi (Aspergillus sp., Rhizopus sp.) to greatly increase their capacity for chemical production. This review highlights the potential of using biotechnology to further develop filamentous bacteria, yeasts, and fungi as alternative chassis strains. Conclusions In this review, we recapitulate the recent progress in the application of filamentous bacteria, yeasts, and fungi as microbial cell factories. Furthermore, emphasis on metabolic engineering strategies involved in cellular tolerance, metabolic engineering, and screening are discussed. Finally, we offer an outlook on advanced techniques for the engineering of filamentous bacteria, yeasts, and fungi.

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An integrated high-throughput strategy for rapid screening of poly(γ-glutamic acid)-producing bacteria

Zeng

Lin

et al. 2013

Appl Microbiol Biotechnol

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Poly(γ-glutamic acid) (γ-PGA) is a promising biomaterial with a wide range of unique applications. To extensively screen γ-PGA-producing bacteria with high yield and different molecular weight, we developed an integrated high-throughput strategy. Firstly, γ-PGA-producing bacteria were selected in a primary screen plate containing a basic dye (neutral red) based on the concentric zone formed through the electrostatic interaction between the dye and the secreted acidic polymer γ-PGA. Then, the isolates were cultured in 50 ml tubes instead of 250 ml flasks. A good correlation of fermentation results in 50 ml tubes and 250 ml flasks was observed. Thirdly, the γ-PGA yield and weight-average molecular weight (M (w)) were simultaneously determined by spectrophotomic assay (UV assay) and neutral red plate assay. The results showed that the diameter of the concentric zone varied among isolates and was negatively correlated with the weight-average molecular weight of γ-PGA. The accuracy of the methods was comparable to that of high-performance liquid chromatography and gel permeation chromatography assay. Lastly, γ-PGA obtained from the target isolates was rapidly identified using thin layer chromatography assay. With this strategy, 13 bacteria with high yield and various molecular weights of γ-PGA from 500 obvious single colonies on the primary screen plate were obtained.

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A high-throughput method for screening of Aspergillus niger mutants with high transglycosylation activity by detecting non-fermentable reducing sugar

Cited by 10 publications

References 19 publications

Non-sterile Submerged Fermentation of Fibrinolytic Enzyme by Marine Bacillus subtilis Harboring Antibacterial Activity With Starvation Strategy

Non-sterile Submerged Fermentation of Fibrinolytic Enzyme by Marine Bacillus subtilis Harboring Antibacterial Activity With Starvation Strategy

Microbial cell factories based on filamentous bacteria, yeasts, and fungi

An integrated high-throughput strategy for rapid screening of poly(γ-glutamic acid)-producing bacteria

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