Recent advances in the development of Aspergillus for protein production

Li, Qinghua; Lu, Jinchang; Zhang, Guoqiang; Liu, Song; Zhou, Jingwen; Du, Guocheng; Chen, Jian

doi:10.1016/j.biortech.2022.126768

Cited by 28 publications

(28 citation statements)

References 112 publications

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“…Despite the fact that the post-genome era of the filamentous fungi has begun, filamentous fungi such as Aspergillus niger have multicellular characteristics [ 47 ]. Compared with single-celled fungi, such as yeast, the growth and development of Aspergillus niger is relatively complex, making molecular genetic manipulation more difficult [ 13 ]. The spore-like propagules exhibit life activities.…”

Section: Discussionmentioning

confidence: 99%

“…The characteristic of hypha is branch growth at the tip and further extension [ 12 ]. In Aspergillus niger , the productions, e.g., proteins, principally appear in the tip of the hyphae [ 12 , 13 ]. Moreover, a study has already indicated that aconidial Aspergillus niger is beneficial for protein biosynthesis and secretion [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, compared with yeast, the genetics and tools for the genetic manipulation of Aspergillus niger are not well developed or comprehensive [ 16 ]. This is mainly due to the complex structure of one of the most prominent characteristics of the hypha [ 13 , 17 ]. A hypha can fuse with other hyphae to form a network called mycelium [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

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A Special Phenotype of Aconidial Aspergillus niger SH2 and Its Mechanism of Formation via CRISPRi

Yao

et al. 2022

JoF

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The complex morphological structure of Aspergillus niger influences its production of proteins, metabolites, etc., making the genetic manipulation and clonal purification of this species increasingly difficult, especially in aconidial Aspergillus niger. In this study, we found that N-acetyl-D-glucosamine (GlcNAc) could induce the formation of spore-like propagules in the aconidial Aspergillus niger SH2 strain. The spore-like propagules possessed life activities such as drug resistance, genetic transformation, and germination. Transcriptomic analysis indicated that the spore-like propagules were resting conidia entering dormancy and becoming more tolerant to environmental stresses. The Dac1 gene and the metabolic pathway of GlcNAc converted to glycolysis are related to the formation of the spore-like propagules, as evidenced by the CRISPRi system, qPCR, and semi-quantitative RT-PCR. Moreover, a method based on the CRISPR-Cas9 tool to rapidly recycle screening tags and recover genes was suitable for Aspergillus niger SH2. To sum up, this suggests that the spore-like propagules are resting conidia and the mechanism of their formation is the metabolic pathway of GlcNAc converted to glycolysis, particularly the Dac1 gene. This study can improve our understanding of the critical factors involved in mechanisms of phenotypic change and provides a good model for researching phenotypic change in filamentous fungi.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Special Phenotype of Aconidial Aspergillus niger SH2 and Its Mechanism of Formation via CRISPRi

Yao

et al. 2022

JoF

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“…A. oryzae has a long history of use in the food industry and is generally recognized as safe (GRAS) by the U.S. Food and Drug Administration (FDA) [ 2 ]. Over the past few decades, A. oryzae has been developed into an efficient cell factory for organic acids, industrial enzymes, and secondary metabolites due to its strong protein secretion ability and post-translational modification characteristics, and it plays an essential role in the fields of food, medicine, feed, and environment [ 3 , 4 ].…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9-Mediated Multiplexed Genome Editing in Aspergillus oryzae

Zhang

et al. 2023

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Aspergillus oryzae has great potential and competitive advantages to be developed as an excellent expression system, owing to its powerful protein secretion ability, complex post-translational modification, and safety characteristics. However, the low efficiency of genetic modification and gene function analysis is an urgent problem to be solved in A. oryzae and other filamentous fungal systems. Therefore, establishing efficient genetic transformation and multiplexed genome editing tools is significant for developing A. oryzae expression systems, and revealing its intrinsic mechanisms. In this study, the high-efficiency transformation of A. oryzae was achieved by optimizing the preparation conditions of protoplasts, and the random editing efficiency of the CRISPR/Cas9 system in A. oryzae for single and double genes reached 37.6% and 19.8%, respectively. With the aid of the selection marker, such as color or resistance, the editing efficiency of single and double genes can reach 100%. Based on the developed CRISPR/Cas9 genome editing method, the heterologous lipase gene (TLL) achieves precise integration at different genetic loci in one step. The efficient and accurate acquisition of positive transformants indicated that the morphological gene yA could be used as a helpful selection marker for genome editing in A. oryzae. In conclusion, the developed system improves the efficiency of transformation and multiplexed genome editing for A. oryzae. It provides a practical method for developing the A. oryzae high-efficiency expression system for heterologous proteins.

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“…Generally, Aspergilli are highly capable of decomposing solid polymers and have been widely used in the fermentation industry for a long time [ 35 , 36 ]. Currently, Aspergilli are used as the host microorganisms for production of these compounds owing to their high productivity of proteins and primary and secondary metabolites [ 37 , 38 , 39 , 40 ]. Aspergilli are used for industrial production of a variety of enzymes, such as amylase, cellulase, glucosidase, hemicellulase, lipase, and phytase from Aspergillus oryzae and Aspergillus niger [ 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 ], and low-molecular-weight compounds such as itaconic acid from Aspergillus terreus [ 53 ], citric acid from A. niger [ 54 ], and kojic acid from A. oryzae [ 55 ].…”

Section: Introductionmentioning

confidence: 99%

Aspergillus Hydrophobins: Physicochemical Properties, Biochemical Properties, and Functions in Solid Polymer Degradation

et al. 2022

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Hydrophobins are small amphipathic proteins conserved in filamentous fungi. In this review, the properties and functions of Aspergillus hydrophobins are comprehensively discussed on the basis of recent findings. Multiple Aspergillus hydrophobins have been identified and categorized in conventional class I and two non-conventional classes. Some Aspergillus hydrophobins can be purified in a water phase without organic solvents. Class I hydrophobins of Aspergilli self-assemble to form amphipathic membranes. At the air–liquid interface, RolA of Aspergillus oryzae self-assembles via four stages, and its self-assembled films consist of two layers, a rodlet membrane facing air and rod-like structures facing liquid. The self-assembly depends mainly on hydrophobin conformation and solution pH. Cys4–Cys5 and Cys7–Cys8 loops, disulfide bonds, and conserved Cys residues of RodA-like hydrophobins are necessary for self-assembly at the interface and for adsorption to solid surfaces. AfRodA helps Aspergillus fumigatus to evade recognition by the host immune system. RodA-like hydrophobins recruit cutinases to promote the hydrolysis of aliphatic polyesters. This mechanism appears to be conserved in Aspergillus and other filamentous fungi, and may be beneficial for their growth. Aspergilli produce various small secreted proteins (SSPs) including hydrophobins, hydrophobic surface–binding proteins, and effector proteins. Aspergilli may use a wide variety of SSPs to decompose solid polymers.

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Recent advances in the development of Aspergillus for protein production

Cited by 28 publications

References 112 publications

A Special Phenotype of Aconidial Aspergillus niger SH2 and Its Mechanism of Formation via CRISPRi

A Special Phenotype of Aconidial Aspergillus niger SH2 and Its Mechanism of Formation via CRISPRi

CRISPR/Cas9-Mediated Multiplexed Genome Editing in Aspergillus oryzae

Aspergillus Hydrophobins: Physicochemical Properties, Biochemical Properties, and Functions in Solid Polymer Degradation

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