Sign in to use this feature.

Years

Between: -

Subjects

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Journals

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Article Types

Countries / Regions

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Search Results (471)

Search Parameters:
Keywords = natural genetic engineering

Order results
Result details
Results per page
Select all
Export citation of selected articles as:
17 pages, 2547 KiB  
Article
A Host Cell Vector Model for Analyzing Viral Protective Antigens and Host Immunity
by Sun-Min Ahn, Jin-Ha Song, Seung-Eun Son, Ho-Won Kim, Gun Kim, Seung-Min Hong, Kang-Seuk Choi and Hyuk-Joon Kwon
Int. J. Mol. Sci. 2025, 26(15), 7492; https://doi.org/10.3390/ijms26157492 - 2 Aug 2025
Viewed by 247
Abstract
Avian influenza A viruses (IAVs) pose a persistent threat to the poultry industry, causing substantial economic losses. Although traditional vaccines have helped reduce the disease burden, they typically rely on multivalent antigens, emphasize humoral immunity, and require intensive production. This study aimed to [...] Read more.
Avian influenza A viruses (IAVs) pose a persistent threat to the poultry industry, causing substantial economic losses. Although traditional vaccines have helped reduce the disease burden, they typically rely on multivalent antigens, emphasize humoral immunity, and require intensive production. This study aimed to establish a genetically matched host–cell system to evaluate antigen-specific immune responses and identify conserved CD8+ T cell epitopes in avian influenza viruses. To this end, we developed an MHC class I genotype (B21)-matched host (Lohmann VALO SPF chicken) and cell vector (DF-1 cell line) model. DF-1 cells were engineered to express the hemagglutinin (HA) gene of clade 2.3.4.4b H5N1 either transiently or stably, and to stably express the matrix 1 (M1) and nucleoprotein (NP) genes of A/chicken/South Korea/SL20/2020 (H9N2, Y280-lineage). Following prime-boost immunization with HA-expressing DF-1 cells, only live cells induced strong hemagglutination inhibition (HI) and virus-neutralizing (VN) antibody titers in haplotype-matched chickens. Importantly, immunization with DF-1 cells transiently expressing NP induced stronger IFN-γ production than those expressing M1, demonstrating the platform’s potential for differentiating antigen-specific cellular responses. CD8+ T cell epitope mapping by mass spectrometry identified one distinct MHC class I-bound peptide from each of the HA-, M1-, and NP-expressing DF-1 cell lines. Notably, the identified HA epitope was conserved in 97.6% of H5-subtype IAVs, and the NP epitope in 98.5% of pan-subtype IAVs. These findings highlight the platform’s utility for antigen dissection and rational vaccine design. While limited by MHC compatibility, this approach enables identification of naturally presented epitopes and provides insight into conserved, functionally constrained viral targets. Full article
(This article belongs to the Special Issue Molecular Research on Immune Response to Virus Infection and Vaccines)
Show Figures

Graphical abstract

48 pages, 3314 KiB  
Review
Applied Microbiology for Sustainable Agricultural Development
by Barbara Sawicka, Piotr Barbaś, Viola Vambol, Dominika Skiba, Piotr Pszczółkowski, Parwiz Niazi and Bernadetta Bienia
Appl. Microbiol. 2025, 5(3), 78; https://doi.org/10.3390/applmicrobiol5030078 - 1 Aug 2025
Viewed by 86
Abstract
Background: Developments in biology, genetics, soil science, plant breeding, engineering, and agricultural microbiology are driving advances in soil microbiology and microbial biotechnology. Material and methods: The literature for this review was collected by searching leading scientific databases such as Embase, Medline/PubMed, Scopus, and [...] Read more.
Background: Developments in biology, genetics, soil science, plant breeding, engineering, and agricultural microbiology are driving advances in soil microbiology and microbial biotechnology. Material and methods: The literature for this review was collected by searching leading scientific databases such as Embase, Medline/PubMed, Scopus, and Web of Science. Results: Recent advances in soil microbiology and biotechnology are discussed, emphasizing the role of microorganisms in sustainable agriculture. It has been shown that soil and plant microbiomes significantly contribute to improving soil fertility and plant and soil health. Microbes promote plant growth through various mechanisms, including potassium, phosphorus, and zinc solubilization, biological nitrogen fixation, production of ammonia, HCN, siderophores, and other secondary metabolites with antagonistic effects. The diversity of microbiomes related to crops, plant protection, and the environment is analyzed, as well as their role in improving food quality, especially under stress conditions. Particular attention was paid to the diversity of microbiomes and their mechanisms supporting plant growth and soil fertility. Conclusions: The key role of soil microorganisms in sustainable agriculture was highlighted. They can support the production of natural substances used as plant protection products, as well as biopesticides, bioregulators, or biofertilizers. Microbial biotechnology also offers potential in the production of sustainable chemicals, such as biofuels or biodegradable plastics (PHA) from plant sugars, and in the production of pharmaceuticals, including antibiotics, hormones, or enzymes. Full article
Show Figures

Figure 1

31 pages, 3024 KiB  
Review
Synthetic and Functional Engineering of Bacteriophages: Approaches for Tailored Bactericidal, Diagnostic, and Delivery Platforms
by Ola Alessa, Yoshifumi Aiba, Mahmoud Arbaah, Yuya Hidaka, Shinya Watanabe, Kazuhiko Miyanaga, Dhammika Leshan Wannigama and Longzhu Cui
Molecules 2025, 30(15), 3132; https://doi.org/10.3390/molecules30153132 - 25 Jul 2025
Viewed by 393
Abstract
Bacteriophages (phages), the most abundant biological entities on Earth, have long served as both model systems and therapeutic tools. Recent advances in synthetic biology and genetic engineering have revolutionized the capacity to tailor phages with enhanced functionality beyond their natural capabilities. This review [...] Read more.
Bacteriophages (phages), the most abundant biological entities on Earth, have long served as both model systems and therapeutic tools. Recent advances in synthetic biology and genetic engineering have revolutionized the capacity to tailor phages with enhanced functionality beyond their natural capabilities. This review outlines the current landscape of synthetic and functional engineering of phages, encompassing both in-vivo and in-vitro strategies. We describe in-vivo approaches such as phage recombineering systems, CRISPR-Cas-assisted editing, and bacterial retron-based methods, as well as synthetic assembly platforms including yeast-based artificial chromosomes, Gibson, Golden Gate, and iPac assemblies. In addition, we explore in-vitro rebooting using TXTL (transcription–translation) systems, which offer a flexible alternative to cell-based rebooting but are less effective for large genomes or structurally complex phages. Special focus is given to the design of customized phages for targeted applications, including host range expansion via receptor-binding protein modifications, delivery of antimicrobial proteins or CRISPR payloads, and the construction of biocontained, non-replicative capsid systems for safe clinical use. Through illustrative examples, we highlight how these technologies enable the transformation of phages into programmable bactericidal agents, precision diagnostic tools, and drug delivery vehicles. Together, these advances establish a powerful foundation for next-generation antimicrobial platforms and synthetic microbiology. Full article
Show Figures

Figure 1

26 pages, 2227 KiB  
Article
Beyond the Hype: Stakeholder Perceptions of Nanotechnology and Genetic Engineering for Sustainable Food Production
by Madison D. Horgan, Christopher L. Cummings, Jennifer Kuzma, Michael Dahlstrom, Ilaria Cimadori, Maude Cuchiara, Colin Larter, Nick Loschin and Khara D. Grieger
Sustainability 2025, 17(15), 6795; https://doi.org/10.3390/su17156795 - 25 Jul 2025
Viewed by 471
Abstract
Ensuring sustainable food systems is an urgent global priority as populations grow and environmental pressures mount. Technological innovations such as genetic engineering (GE) and nanotechnology (nano) have been promoted as promising pathways for achieving greater sustainability in agriculture and food production. Yet, the [...] Read more.
Ensuring sustainable food systems is an urgent global priority as populations grow and environmental pressures mount. Technological innovations such as genetic engineering (GE) and nanotechnology (nano) have been promoted as promising pathways for achieving greater sustainability in agriculture and food production. Yet, the sustainability of these technologies is not defined by technical performance alone; it hinges on how they are perceived by key stakeholders and how well they align with broader societal values. This study addresses the critical question of how expert stakeholders evaluate the sustainability of GE and nano-based food and agriculture (agrifood) products. Using a multi-method online platform, we engaged 42 experts across academia, government, industry, and NGOs in the United States to assess six real-world case studies—three using GE and three using nano—across ten different dimensions of sustainability. We show that nano-based products were consistently rated more favorably than their GE counterparts in terms of environmental, economic, and social sustainability, as well as across ethical and societal dimensions. Like prior studies, our results reveal that stakeholders see meaningful distinctions between nanotechnology and biotechnology, likely due to underlying value-based concerns about animal welfare, perceived naturalness, or corporate control of agrifood systems. The fruit coating and flu vaccine—both nano-enabled—received the most positive ratings, while GE mustard greens and salmon were the most polarizing. These results underscore the importance of incorporating stakeholder perspectives in technology assessment and innovation governance. These results also suggest that responsible innovation efforts in agrifood systems should prioritize communication, addressing meaningful societal needs, and the contextual understanding of societal values to build trust and legitimacy. Full article
(This article belongs to the Special Issue Food Science and Engineering for Sustainability)
Show Figures

Figure 1

15 pages, 1351 KiB  
Review
Unraveling the Complexity of Plant Trichomes: Models, Mechanisms, and Bioengineering Strategies
by Tiantian Chen, Yanfei Ma and Jiyan Qi
Int. J. Mol. Sci. 2025, 26(14), 7008; https://doi.org/10.3390/ijms26147008 - 21 Jul 2025
Viewed by 431
Abstract
Trichomes—microscopic appendages on the plant epidermis—play vital roles as both protective barriers and specialized biosynthetic factories. Acting as the first line of defense against environmental stressors, they also produce a wide range of pharmaceutically valuable secondary metabolites. This mini-review highlights recent advances in [...] Read more.
Trichomes—microscopic appendages on the plant epidermis—play vital roles as both protective barriers and specialized biosynthetic factories. Acting as the first line of defense against environmental stressors, they also produce a wide range of pharmaceutically valuable secondary metabolites. This mini-review highlights recent advances in understanding the development, structure, and function of trichomes, with a focus on glandular secretory trichomes (GSTs) in key species such as Artemisia annua and Solanum lycopersicum. We explore how insights from these systems are driving innovation in plant synthetic biology, including modular genetic engineering and metabolic channeling strategies. These breakthroughs are paving the way for scalable, plant-based platforms to produce high-value compounds. By integrating molecular mechanisms with emerging technologies, this review outlines a forward-looking framework for leveraging trichomes in sustainable agriculture, natural product discovery, and next-generation biomanufacturing. Full article
Show Figures

Figure 1

67 pages, 4242 KiB  
Review
Bioengineering Outer-Membrane Vesicles for Vaccine Development: Strategies, Advances, and Perspectives
by Ayesha Zahid, Hazrat Ismail, Jennifer C. Wilson and I. Darren Grice
Vaccines 2025, 13(7), 767; https://doi.org/10.3390/vaccines13070767 - 20 Jul 2025
Viewed by 927
Abstract
Outer-membrane vesicles (OMVs), naturally secreted by Gram-negative bacteria, have gained recognition as a versatile platform for the development of next-generation vaccines. OMVs are essential contributors to bacterial pathogenesis, horizontal gene transfer, cellular communication, the maintenance of bacterial fitness, and quorum sensing. Their intrinsic [...] Read more.
Outer-membrane vesicles (OMVs), naturally secreted by Gram-negative bacteria, have gained recognition as a versatile platform for the development of next-generation vaccines. OMVs are essential contributors to bacterial pathogenesis, horizontal gene transfer, cellular communication, the maintenance of bacterial fitness, and quorum sensing. Their intrinsic immunogenicity, adjuvant properties, and scalability establish OMVs as potent tools for combating infectious diseases and cancer. Recent advancements in genetic engineering and biotechnology have further expanded the utility of OMVs, enabling the incorporation of multiple epitopes and antigens from diverse pathogens. These developments address critical challenges such as antigenic variability and co-infections, offering broader immune coverage and cost-effective solutions. This review explores the unique structural and immunological properties of OMVs, emphasizing their capacity to elicit robust immune responses. It critically examines established and emerging engineering strategies, including the genetic engineering of surface-displayed antigens, surface conjugation, glycoengineering, nanoparticle-based OMV engineering, hybrid OMVs, and in situ OMV production, among others. Furthermore, recent advancements in preclinical research on OMV-based vaccines, including synthetic OMVs, OMV-based nanorobots, and nanodiscs, as well as emerging isolation and purification methods, are discussed. Lastly, future directions are proposed, highlighting the potential integration of synthetic biology techniques to accelerate research on OMV engineering. Full article
(This article belongs to the Special Issue Bioengineering Strategies for Developing Vaccines)
Show Figures

Graphical abstract

24 pages, 2860 KiB  
Review
Recent Evidence for Orthobiologics Combined with Hydrogels for Joint Tissue Regeneration: Focus on Osteoarthritis
by Carola Cavallo, Giovanna Desando, Martina D’Alessandro, Brunella Grigolo and Livia Roseti
Gels 2025, 11(7), 551; https://doi.org/10.3390/gels11070551 - 17 Jul 2025
Viewed by 363
Abstract
Osteoarthritis is a significant global problem, causing pain and limitations, and contributing to socioeconomic expenses. The etiopathogenesis of this disease encloses genetic, biological, and mechanical aspects. Regenerative medicine, utilizing tissue engineering, has opened the way to new therapeutic approaches employing various orthobiologics. Combined [...] Read more.
Osteoarthritis is a significant global problem, causing pain and limitations, and contributing to socioeconomic expenses. The etiopathogenesis of this disease encloses genetic, biological, and mechanical aspects. Regenerative medicine, utilizing tissue engineering, has opened the way to new therapeutic approaches employing various orthobiologics. Combined with hydrogels, these compounds may represent a notable option for treating degenerative and inflammatory lesions in OA. The review reports on the main orthobiologics used in preclinical and clinical studies, as well as their association with various types of natural and synthetic hydrogels. Research may increasingly focus on tailored therapies adjusted to suit the joint involved and the severity of the pathology encountered in each patient. Full article
Show Figures

Figure 1

43 pages, 7260 KiB  
Article
A Solution Method for Non-Linear Underdetermined Equation Systems in Grounding Grid Corrosion Diagnosis Based on an Enhanced Hippopotamus Optimization Algorithm
by Jinhe Chen, Jianyu Qi, Yiyang Ao, Keying Wang and Xin Song
Biomimetics 2025, 10(7), 467; https://doi.org/10.3390/biomimetics10070467 - 16 Jul 2025
Viewed by 466
Abstract
As power grids scale and aging assets edge toward obsolescence, grounding grid corrosion has become a critical vulnerability. Conventional diagnosis must fit high-dimensional electrical data to a physical model, typically yielding a nonlinear under-determined system fraught with computational burden and uncertainty. We propose [...] Read more.
As power grids scale and aging assets edge toward obsolescence, grounding grid corrosion has become a critical vulnerability. Conventional diagnosis must fit high-dimensional electrical data to a physical model, typically yielding a nonlinear under-determined system fraught with computational burden and uncertainty. We propose the Enhanced Biomimetic Hippopotamus Optimization (EBOHO) algorithm, which distills the river-dwelling hippo’s ecological wisdom into three synergistic strategies: a beta-function herd seeding that replicates the genetic diversity of juvenile hippos diffusing through wetlands, an elite–mean cooperative foraging rule that echoes the way dominant bulls steer the herd toward nutrient-rich pastures, and a lens imaging opposition maneuver inspired by moonlit water reflections that spawn mirror candidates to avert premature convergence. Benchmarks on the CEC 2017 suite and four classical design problems show EBOHO’s superior global search, robustness, and convergence speed over numerous state-of-the-art meta-heuristics, including prior hippo variants. An industrial case study on grounding grid corrosion further confirms that EBOHO swiftly resolves the under-determined equations and pinpoints corrosion sites with high precision, underscoring its promise as a nature-inspired diagnostic engine for aging power system infrastructure. Full article
Show Figures

Figure 1

13 pages, 1764 KiB  
Article
Surface Display of Avian H5 and H9 Hemagglutinin Antigens on Non-Genetically Modified Lactobacillus Cells for Bivalent Oral AIV Vaccine Development
by Fuyi Liu, Jingbo Chang, Jingqi Huang, Yuping Liao, Xiaonan Deng, Tingting Guo, Jian Kong and Wentao Kong
Microorganisms 2025, 13(7), 1649; https://doi.org/10.3390/microorganisms13071649 - 11 Jul 2025
Viewed by 354
Abstract
A novel bivalent oral vaccine candidate against H5N1 and H9N2 avian influenza virus (AIV) was developed using Lactobacillus surface display technology without genetic modification. The hemagglutinin subunit 1 (HA1) antigens from both subtypes were fused to the surface layer-binding domain of Lactobacillus crispatus [...] Read more.
A novel bivalent oral vaccine candidate against H5N1 and H9N2 avian influenza virus (AIV) was developed using Lactobacillus surface display technology without genetic modification. The hemagglutinin subunit 1 (HA1) antigens from both subtypes were fused to the surface layer-binding domain of Lactobacillus crispatus K313, expressed in Escherichia coli, and purified. Wild-type Lactobacillus johnsonii H31, isolated from chicken intestine, served as a delivery vehicle by adsorbing and stably displaying the HA1 proteins on its surface. This approach eliminates the need for bacterial engineering while utilizing lactobacilli’s natural capacity to protect surface-displayed antigens, as evidenced by HA1’s protease resistance. Mouse immunization studies demonstrated induction of strong systemic IgG and mucosal IgA responses against both H5N1 and H9N2 HA1. The system offers several advantages, including safety through non-GMO probiotics, potential for multivalent vaccine expansion, and intrinsic antigen protection by lactobacilli. These findings suggest this platform could enable development of cost-effective, multivalent AIV vaccines. Full article
(This article belongs to the Section Food Microbiology)
Show Figures

Figure 1

37 pages, 3498 KiB  
Review
Pigments from Microorganisms: A Sustainable Alternative for Synthetic Food Coloring
by Akshay Chavan, Jaya Pawar, Umesh Kakde, Mekala Venkatachalam, Mireille Fouillaud, Laurent Dufossé and Sunil Kumar Deshmukh
Fermentation 2025, 11(7), 395; https://doi.org/10.3390/fermentation11070395 - 10 Jul 2025
Viewed by 1057
Abstract
Microbial pigments are gaining acceptance as a green, sustainable substitute for synthetic food pigments due to growing health issues and their adverse health impacts. This review provides an overview of the potential of microbial pigments as natural food colorants and the advantages of [...] Read more.
Microbial pigments are gaining acceptance as a green, sustainable substitute for synthetic food pigments due to growing health issues and their adverse health impacts. This review provides an overview of the potential of microbial pigments as natural food colorants and the advantages of microbial pigments over synthetic pigments. Microbial pigments are a natural source of color with medicinal properties like anticancer, antimicrobial, and antioxidant activity. Important pigments covered are astaxanthin, phycocyanin, prodigiosin, riboflavin, β-carotene, violacein, melanin, and lycopene, and their microbial origins and characteristics. The review also covers commercial production of microbial pigments, i.e., strain development and fermentation processes. Microbial pigments also find extensive applications in food industries, including preservatives for food. Also covered are their pharmacological activity and other applications, such as in the formation of nanoparticles. Finally, the challenges and future directions of microbial pigment production are covered, including the need for cost-effective production, regulatory acceptability, and the potential of genetic engineering and fermentation-based technologies to enhance pigment yield and quality. Full article
(This article belongs to the Special Issue Metabolic Engineering in Microbial Synthesis)
Show Figures

Figure 1

17 pages, 1886 KiB  
Review
Random Insertion Reporter Gimmicks Powered by Cut-and-Paste DNA Transposons
by Yamato Kasahara, Kentaro Semba, Shinya Watanabe and Kosuke Ishikawa
Biomedicines 2025, 13(7), 1682; https://doi.org/10.3390/biomedicines13071682 - 9 Jul 2025
Viewed by 381
Abstract
Transposons are mobile genetic elements capable of moving within the genome. Leveraging this property—particularly the cut-and-paste mechanism of DNA transposons—has enabled the development of technologies for inserting exogenous DNA fragments into host genomes. While targeted integration is a key goal for therapeutic applications, [...] Read more.
Transposons are mobile genetic elements capable of moving within the genome. Leveraging this property—particularly the cut-and-paste mechanism of DNA transposons—has enabled the development of technologies for inserting exogenous DNA fragments into host genomes. While targeted integration is a key goal for therapeutic applications, this review highlights the value of their intrinsic randomness. By combining the ability to freely design the DNA cargo with the stochastic nature of transposon integration, it becomes possible to generate highly sensitive reporter cells. These can be used to efficiently identify functional markers, uncover novel signaling pathways, and establish innovative platforms for drug screening. As more subfamilies of transposons become available for research use, their complementary biases may enhance the coverage and diversity of genome-wide screening approaches. Although inherently unpredictable, this strategy embraces randomness as a strength, and we propose that it holds great promise for driving new advances in biology, cellular engineering, and medical research. Full article
(This article belongs to the Special Issue Gene Delivery and Gene Editing)
Show Figures

Figure 1

28 pages, 18319 KiB  
Review
Influence of Scaffold Structure and Biomimetic Properties on Adipose Stem Cell Homing in Personalized Reconstructive Medicine
by Doina Ramona Manu, Diana V. Portan, Monica Vuţă and Minodora Dobreanu
Biomimetics 2025, 10(7), 438; https://doi.org/10.3390/biomimetics10070438 - 3 Jul 2025
Viewed by 597
Abstract
Human adipose stem cells (ASCs) are multipotent cells expressing mesenchymal stem cell (MSC) markers that are capable of multilineage differentiation and secretion of bioactive factors. Their “homing” to injured tissues is mediated by chemokines, cytokines, adhesion molecules, and signaling pathways. Enhancing ASC homing [...] Read more.
Human adipose stem cells (ASCs) are multipotent cells expressing mesenchymal stem cell (MSC) markers that are capable of multilineage differentiation and secretion of bioactive factors. Their “homing” to injured tissues is mediated by chemokines, cytokines, adhesion molecules, and signaling pathways. Enhancing ASC homing is critical for improving regenerative therapies. Strategies include boosting chemotactic signaling, modulating immune responses to create a supportive environment, preconditioning ASCs with hypoxia or mechanical stimuli, co-culturing with supportive cells, applying surface modifications or genetic engineering, and using biomaterials to promote ASC recruitment, retention, and integration at injury sites. Scaffolds provide structural support and a biomimetic environment for ASC-based tissue regeneration. Natural scaffolds promote adhesion and differentiation but have mechanical limitations, while synthetic scaffolds offer tunable properties and controlled degradation. Functionalization with bioactive molecules improves the regenerative outcomes of different tissue types. Ceramic-based scaffolds, due to their strength and bioactivity, are ideal for bone healing. Composite scaffolds, combining polymers, ceramics, or metals, further optimize mechanical and biological properties, supporting personalized regenerative therapies. This review integrates concepts from cell biology, biomaterials science, and regenerative medicine to offer a comprehensive understanding of ASC homing and its impact on tissue engineering and clinical applications. Full article
(This article belongs to the Section Biomimetics of Materials and Structures)
Show Figures

Figure 1

32 pages, 1834 KiB  
Review
Regulation of Riboflavin Biosynthesis in Microorganisms and Construction of the Advanced Overproducers of This Vitamin
by Justyna Ruchala, Alicja Najdecka, Dominik Wojdyla, Wen Liu and Andriy Sibirny
Int. J. Mol. Sci. 2025, 26(13), 6243; https://doi.org/10.3390/ijms26136243 - 28 Jun 2025
Viewed by 674
Abstract
Riboflavin (vitamin B2) is an essential micronutrient required for all living organisms. It is naturally synthesized by plants and most microorganisms, including the bacterium Bacillus subtilis, the filamentous fungus Ashbya gossypii, and the yeast Candida famata—all of which [...] Read more.
Riboflavin (vitamin B2) is an essential micronutrient required for all living organisms. It is naturally synthesized by plants and most microorganisms, including the bacterium Bacillus subtilis, the filamentous fungus Ashbya gossypii, and the yeast Candida famata—all of which are known to be riboflavin overproducers. The choice of production organism in industrial applications depends on factors such as yield, ease of cultivation, and the availability of genetic tools. As a result, several microorganisms are commonly used, and their relative prominence can shift over time with advances in metabolic engineering and process optimization. This review presents a comparative analysis of riboflavin biosynthesis across prokaryotic and eukaryotic systems, with a particular focus on regulatory mechanisms governing flavinogenesis. Special attention is given to recent advances in metabolic engineering strategies, including the application of CRISPR/Cas9 genome editing in Bacillus subtilis and Ashbya gossypii. In yeast systems, significant improvements in riboflavin production have been achieved primarily through the manipulation of transcriptional regulators (e.g., SEF1, SFU1, TUP1) and metabolic genes. The role of other important genes (PRS3, ADE4, ZWF1, GND1, RFE1, VMA1, etc.) in riboflavin overproduction in C. famata is described. The review also explores the use of alternative, low-cost feedstocks—including lignocellulosic hydrolysates and dairy by-products—to support more sustainable and economically viable riboflavin production. Although considerable progress has been achieved in genetic optimization and bioprocess development, further work is required to fine-tune metabolic flux and maximize riboflavin synthesis, particularly under industrial conditions. This review highlights key opportunities for future research aimed at refining metabolic interventions and expanding the use of renewable substrates for environmentally sustainable riboflavin production. Full article
(This article belongs to the Special Issue New Advances in Metabolic Engineering and Synthetic Biology of Yeasts)
Show Figures

Figure 1

20 pages, 5356 KiB  
Article
Structure–Function Analysis of the Steroid-Hydroxylating Cytochrome P450 109 (CYP109) Enzyme Family
by Siphesihle M. Msweli, Tiara Padayachee, Thembeka Khumalo, David R. Nelson, David C. Lamb and Khajamohiddin Syed
Int. J. Mol. Sci. 2025, 26(13), 6219; https://doi.org/10.3390/ijms26136219 - 27 Jun 2025
Viewed by 433
Abstract
Steroids are found in bacteria and eukaryotes, and genes potentially encoding steroid metabolic enzymes have also been identified in giant viruses. For decades, hydroxylated steroids have been utilized in medicine to treat various human diseases. The hydroxylation of steroids can be achieved using [...] Read more.
Steroids are found in bacteria and eukaryotes, and genes potentially encoding steroid metabolic enzymes have also been identified in giant viruses. For decades, hydroxylated steroids have been utilized in medicine to treat various human diseases. The hydroxylation of steroids can be achieved using microbial enzymes, especially cytochrome P450 monooxygenases (CYPs/P450s) and is well documented. Understanding the structural determinants that govern the regio- and stereoselectivity of steroid hydroxylation by P450s is essential in order to fully exploit their potential. Herein, we present a comprehensive analysis of the steroid-hydroxylating CYP109 family across the domains of life and delineate the structural determinants that govern steroid hydroxylation. Data mining, annotation, and phylogenetic analysis revealed that CYP109 family members are highly populated in bacteria, and indeed, these members passed from bacteria to archaea by horizontal gene transfer, leading to the evolution of P450s in archaea. Analysis of twelve CYP109 crystal structures revealed large, flexible, and dynamic active site cavities that can accommodate multiple ligands. The correct positioning and orientation of the steroid in the active site cavity and the nature of the C17 substituent on the steroid molecule influence catalysis. In an analogous fashion to the CYP107 family, the amino acid residues within the CYP109 binding pocket involve hydrophilic and hydrophobic interactions, influencing substrate orientations and anchoring and determining the site of hydroxylation and catalytic activity. A handful of amino acids, such as Val84, Val292, and Ser387 in CYP109B4, have been found to play a role in determining the catalytic regiospecificity, and a single amino acid, such as Arg74 in CYP109A2, has been found to be essential for the enzymatic activity. This work serves as a reference for the precise understanding of CYP109 structure–function relationships and for P450 enzymes in general. The findings will guide the genetic engineering of CYP109 enzymes to produce valuable steroid molecules of medicinal and biotechnological importance. Full article
(This article belongs to the Section Biochemistry)
Show Figures

Figure 1

21 pages, 1610 KiB  
Review
Plant Transformation and Genome Editing for Precise Synthetic Biology Applications
by Sharathchandra Kambampati, Pankaj K. Verma and Madhusudhana R. Janga
SynBio 2025, 3(3), 9; https://doi.org/10.3390/synbio3030009 - 27 Jun 2025
Viewed by 794
Abstract
Synthetic biology (SynBio) is an emerging interdisciplinary field that applies engineering principles to the design and construction of novel biological systems or the redesign of existing natural systems for new functions. As autotrophs with complex cellular architectures, plants possess inherent capabilities to serve [...] Read more.
Synthetic biology (SynBio) is an emerging interdisciplinary field that applies engineering principles to the design and construction of novel biological systems or the redesign of existing natural systems for new functions. As autotrophs with complex cellular architectures, plants possess inherent capabilities to serve as “living factories” for SynBio applications. Recent advancements in genetic engineering, genome editing, and transformation techniques are improving the precision and programmability of plant systems. Innovations, such as CRISPR systems, prime editing strategies, and in planta and nanoparticle-mediated delivery, are expanding the SynBio toolkit for plants. However, the efficient delivery of genetic constructs remains a barrier due to plant systems’ complexity. To address these limitations, SynBio is increasingly integrating iterative Design–Build–Test–Learn (DBTL) cycles, standardization, modular DNA assembly systems, and plant-optimized toolkits to enable predictable trait engineering. This review explores the technological foundations of plant SynBio, including genome editing and transformation methods, and examines their integration into engineered systems. Applications, such as biofuel production, pharmaceutical biosynthesis, and agricultural innovation, are highlighted, along with their ethical, technical, and regulatory challenges. Ultimately, SynBio could offer a transformative path toward sustainable solutions, provided it continues to align technological advances with public interest and global sustainability goals. Full article
Show Figures

Figure 1

Back to TopTop