Potential for Producing Fourth-generation Biological Products via Genetic Engineering of Filamentous Fungi

Peeter Vivi^*
*Correspondence: Peeter Vivi, Department of General Practice, Federal University of Goias, Km-0, Goiânia P.O. Box 131, GO, Brazil, Email:

Keywords

Biotechnology • Environments • Cellulase production

Introduction

The metabolic diversity of filamentous fungi allows them to produce a wide range of secondary metabolites, including antibiotics, enzymes, and organic acids. This makes them valuable organisms for biotechnological applications. Additionally, their ability to grow on inexpensive and abundant substrates makes them economically attractive for industrial-scale production. Enzymes produced by filamentous fungi are widely used in various industries, including food, pharmaceuticals, and biofuels. Genetic engineering can enhance enzyme production by optimizing gene expression, secretion pathways, and enzyme stability. For example, the production of cellulases and hemicellulases for biofuel production can be significantly improved through genetic modifications. Trichoderma reesei is a well-known producer of cellulases, enzymes that break down cellulose into glucose. Genetic engineering efforts have focused on increasing cellulase production by overexpressing key cellulase genes and eliminating regulatory bottlenecks. CRISPR-Cas9 has been used to create targeted deletions of genes involved in negative regulation, resulting in higher enzyme yields. Filamentous fungi are a rich source of antibiotics, and genetic engineering can enhance the production of these valuable compounds. By manipulating biosynthetic gene clusters, researchers can increase antibiotic yields and create novel antibiotics with improved properties. Penicillium chrysogenum is the source of penicillin, the first antibiotic discovered. Genetic engineering has been used to improve penicillin production by optimizing the expression of biosynthetic genes and introducing genes from other organisms to enhance the biosynthetic pathway. This has led to strains with significantly higher penicillin yields. Organic acids, such as citric acid, fumaric acid, and lactic acid, are important industrial chemicals. Filamentous fungi can be engineered to produce these acids in high yields by optimizing metabolic pathways and improving tolerance to acid production [1,2].

Literature Review

Aspergillus niger is widely used for citric acid production. Genetic engineering efforts have focused on increasing the flux through the citric acid biosynthetic pathway and improving the strain's tolerance to high citric acid concentrations. Overexpression of key enzymes in the pathway, such as citrate synthase, has led to improved citric acid production. The ability to produce novel metabolites through synthetic biology and metabolic engineering is a hallmark of fourthgeneration biological products. By constructing new biosynthetic pathways, researchers can create fungi that produce valuable compounds not naturally found in these organisms. Researchers have successfully engineered filamentous fungi to produce natural products from other organisms. For example, the production of the anti-cancer compound taxol, originally derived from the yew tree, has been achieved in Aspergillus species through the introduction of the entire taxol biosynthetic pathway. This demonstrates the potential of filamentous fungi as versatile platforms for producing high-value compounds [3,4].

Discussion

While the potential for producing fourth-generation biological products via genetic engineering of filamentous fungi is immense, several challenges need to be addressed. Continuous efforts are needed to improve fungal strains for industrial applications. This includes optimizing growth conditions, metabolic pathways, and product yields. Genetically engineered organisms face regulatory scrutiny. Ensuring the safety and environmental impact of these organisms is critical for their acceptance and commercialization. Transitioning from laboratory-scale production to industrial-scale production requires overcoming technical and economic challenges. Efficient fermentation processes and downstream processing are essential for commercial viability. Ensuring the safe use and containment of genetically modified fungi is crucial to prevent unintended environmental release and cross-contamination. Developing new genetic tools and techniques to enhance the precision and efficiency of genetic modifications in filamentous fungi. Expanding the repertoire of metabolic engineering strategies to create more efficient and versatile fungal cell factories. Leveraging synthetic biology to design and construct novel biosynthetic pathways and regulatory circuits for the production of high-value compounds. Integrating systems biology approaches to understand and optimize complex metabolic networks in filamentous fungi [5,6].

Conclusion

The genetic engineering of filamentous fungi holds significant promise for the production of fourth-generation biological products. With their diverse metabolic capabilities, ease of genetic manipulation, and potential for highyield production, filamentous fungi are poised to become key players in sustainable biotechnology. By addressing current challenges and leveraging advanced genetic and synthetic biology techniques, researchers can unlock the full potential of these remarkable organisms, paving the way for innovative and sustainable biological products. Techniques such as Agrobacteriummediated transformation, electroporation, and protoplast fusion are commonly used to introduce foreign DNA into fungal cells. The CRISPR-Cas9 system has revolutionized genetic engineering by allowing precise and targeted modifications. It has been successfully applied to filamentous fungi for gene editing and regulation. Overexpression of genes can enhance the production of desired metabolites, while gene knockout can eliminate unwanted pathways and by-products. The use of synthetic biology approaches, including the design and construction of synthetic gene circuits, enables the creation of novel metabolic pathways and the optimization of existing ones.

Acknowledgement

None.

Conflict of Interest

None.

References

1. Nevalainen, Helena and Robyn Peterson. "Making recombinant proteins in filamentous fungi-are we expecting too much?." Front Microbiol 5 (2014): 76292.

   Google Scholar, Crossref, Indexed at

2. Demain, Arnold L. "Small bugs, big business: The economic power of the microbe." Biotechnology advances 18 (2000): 499-514.

   Google Scholar, Crossref, Indexed at

3. Wang, Qin, Chao Zhong and Han Xiao. "Genetic engineering of filamentous fungi for efficient protein expression and secretion." Front Bioeng Biotechnol 8 (2020): 293.

   Google Scholar, Crossref, Indexed at

4. Corrêa, Bianca, Vanessa da Silveira Duarte, Daniela Milanez Silva and Gabriel Moura Mascarin, et al. "Comparative analysis of blastospore production and virulence of Beauveria bassiana and Cordyceps fumosorosea against soybean pests." BioControl 65 (2020): 323-337.

   Google Scholar, Crossref, Indexed at

5. Woo, Sheridan L., Rosa Hermosa, Matteo Lorito and Enrique Monte. "Trichoderma: A multipurpose, plant-beneficial microorganism for eco-sustainable agriculture." Nat Rev Microbiol 21 (2023): 312-326.

   Google Scholar, Crossref, Indexed at

6. Joop, Gerrit and Andreas Vilcinskas. "Coevolution of parasitic fungi and insect hosts." Zoology 119 (2016): 350-358.

   Google Scholar, Crossref, Indexed at