Biotechnological Applications Research Articles

Genome-based analysis of biosynthetic potential from antimycotic Streptomyces rochei strain A144.

Streptomyces rochei is a species of Streptomyces with a diverse range of biological activities. S. rochei strain A144 was isolated from desert soils and exhibits antagonistic activity against several plant pathogenic fungi. The genome of S. rochei A144 was sequenced and revealed the presence of one linear chromosome and one plasmid. The chromosome length was found to be 8,085,429bp, with a GC content of 72.62%, while the Plas1 length was 177,399bp, with a GC content of 69.08%. Comparative genomics was employed to analyse the S. rochei group. There is a high degree of collinearity between the genomes of S. rochei strains. Based on pan-genome analysis, S. rochei has 10,315 gene families, including 4051 core and 2322 unique genes. AntiSMASH was used to identify the gene clusters for secondary metabolites, identifying 33 secondary metabolite genes on the A144 genome. Among them, 18 clusters were found to be >70% identical to known biosynthetic gene clusters (BGCs), indicating that A144 has the potential to synthesize secondary metabolites. The majority of the BGCs were found to be conserved within the S. rochei group, including those encoding polyketide synthases (PKS), terpenes, non-ribosomal peptide synthetases (NRPS), other ribosomally synthesised and post-translationally modified peptides (RiPP), nicotianamine-iron transporters, lanthipeptides, and a few other types. The S. rochei group can be a potential genetic source of useful secondary metabolites with applications in medicine and biotechnology.

Droplet-Based Microfluidic Photobioreactor as a Growth Optimization Tool for Cyanobacteria and Microalgae

Microalgae and cyanobacteria are photosynthetic microorganisms with significant biotechnological potential for the production of bioactive compounds, making them a promising resource for diverse industrial applications. This study presents the development and validation of a modular, droplet-based microfluidic photobioreactor (µPBR) designed for high-throughput screening and cultivation under controlled light conditions. The µPBR, based on polytetrafluoroethylene (PTFE) tubing and a 4-channel LED illumination system, enables precise modulation of light intensity, wavelength, and photoperiod, facilitating dose–response experiments. Synechococcus elongatus UTEX 2973 and Chlorella vulgaris were used to demonstrate the system’s capacity to support photosynthetic growth under various conditions. The results indicate that continuous illumination, particularly under blue and mixed blue-red light, promotes higher autofluorescence and chlorophyll a content in cyanobacteria Synechococcus elongatus UTEX2973, while Chlorella vulgaris achieved optimal growth under a 16:8 light-dark cycle with moderate light intensity. This µPBR offers not only a flexible, scalable platform for optimizing growth parameters but also allows for the investigation of highly resolved dose response screenings of environmental stressors such as salinity. The presented findings highlight its potential for advancing microalgal biotechnology research and applications.

Interfacing Complex Coacervates with Natural Cells

Coacervates have been investigated as protocells or synthetic cells, as well as subcellular compartments for the creation of new materials, thus bridging the gap between living and non‐living systems in materials science, synthetic biology, and bioengineering. Given the design flexibility and simplicity of coacervates, along with the functionality and complexity of natural cells, the interfacing of complex coacervates with natural cells is considered significant for various biotechnological and biomedical applications. In this review, the fundamental mechanisms and underlying theories of coacervate systems are introduced. Recent efforts to interface coacervates with natural cells are summarized in three key scenarios: (i) the integration of coacervates with natural cell components for the living material assembly into protocells; (ii) communication between therapeutic synthetic cells and natural cells for drug delivery and cell repair; and (iii) the formation of intracellular condensates for metabolic regulation, followed by the regulation of their phase transitions for pathological elucidation. Finally, the potential of coacervate‐natural cell interfaces is discussed in the context of developing living/synthetic cell constructs, creating precise disease therapy strategies, and advancing programmable metabolic engineering networks.

Bioengineering Bacillus spp. for Sustainable Crop Production: Recent Advances and Resources for Biotechnological Applications

AbstractThe goal of sustainable agriculture is to meet the rising need for food, while minimizing adverse impacts on the environment, protecting natural resources, and ensuring agricultural output over the long term. The pressing need to increase agricultural yield through sustainable agriculture is being emphasized. Several Bacillus species have been used as commercial biopesticides since they can act against plant pathogens by potentially suppressing them. At the same time, they can act as plant growth-promoting rhizobacteria and are known for their diverse characteristics and beneficial properties, making them potential candidates for use sustainable crop production programs. Knowledge of genetic information opens the door of possibility for understanding the way these microorganisms behave. By applying biotechnological tools to Bacillus, strategies can be adopted for the purpose of increasing the yield of crops and managing pests and pathogens that infect them. In this review, we identify the genes in the most significant Bacillus spp. that contribute to plant improvement. The most important biotechnological tools and advance computational approaches are described to provide an extended vision on this topic. However, increasing the crop production through application of beneficial microbial strains requires a multifaceted approach that considers ecological, economic, and social aspects. By implementing these strategies and practices, we can work towards a sustainable and resilient agricultural system that meets the growing food demand, while preserving the environment for future generations.

Chemical diversity of cyanobacterial natural products.

Covering: 2010 to 2023Cyanobacterial natural products are a diverse group of molecules with promising biotechnological applications. This review examines the chemical diversity of 995 cyanobacterial metabolites reported from 2010 to 2023. A computational analysis using similarity networking was applied to visualize the chemical space and to compare the diversity of cyanobacterial metabolites among taxonomic orders and environmental sources. Key examples are highlighted, detailing their sources, biological activities, and discovery processes.

Characteristics of deep-sea microbial cellulases: key determinants of the ultimate fate of plant biomass on Earth

AbstractLand plants, especially those with significant woody biomass, represent the largest source of biomass on Earth, making the biodegradation of lignocellulosic materials critical to understanding the global carbon cycle. Cellulose, a major component of lignocellulose, is notoriously resistant to degradation due to its highly crystalline structure. While the degradation of cellulose by terrestrial microbes has been extensively studied, the mechanisms of cellulose degradation in deep-sea environments remain largely unexplored. The deep-sea ecosystem depends on organic matter, such as cellulose, that is synthesized in terrestrial environments and surface waters and descends to the deep sea. Recent studies suggest that a significant amount of cellulose is likely to reach the deep sea. Here, we present an in-depth study of cellulases from a novel deep-sea γ-proteobacterial strain TOYAMA8, isolated from Toyama Bay, Japan, using Surface Pitting Observation Technology (SPOT), a highly sensitive assay for enzymatic cellulose hydrolysis. The cellulases of strain TOYAMA8 show similarities to those of a previously reported deep-sea cellulolytic microbe, Marinagarivorans cellulosilyticus strain GE09. Genomic and transcriptomic analyses of these strains reveal novel cellulase genes and mechanisms that differ from terrestrial counterparts, shedding light on the unique adaptations of deep-sea microbes to recalcitrant biomass. In particular, these strains produce high-molecular-weight cellulases with unique domain architectures, likely optimized for membrane anchoring, which prevents enzyme diffusion and ensures efficient localized activity. Our findings provide critical insights into the microbial cellulose degradation in the deep sea, highlighting its role in the fate of organic carbon and the potential for biotechnological applications in biorefineries.

Direct conversion of dissolved air flotation sludge to mannosylerythritol lipids by Moesziomyces aphidis

AbstractDissolved air flotation (DAF) sludge from wastewater treatment plants is often a lipid‐rich waste stream, which is now mostly being converted to biogas. Due to its high carbon content, DAF sludge has great potential in biotechnological applications. MEL production with floated grease from a sauce producer as substrate resulted in the accumulation of 1.1 ± 0.1 g/L MEL. With enhanced aeration through the use of baffled shaking flasks, MEL titres increased up to 9.7 ± 0.5 g/L. An adaptation period of 9 days was observed, after which the specific productivity reached 0.20 g/L.h. This study shows for the first time biosurfactant production from DAF sludge, with promising titres. Further research should focus on reducing the lag phase to increase productivity.

Global Profiling of Protein Phosphorylation, Acetylation, and β-Hydroxybutyrylation in Nannochloropsis oceanica.

Protein post-translational modifications (PTMs) regulate protein functions but remain poorly characterized in Nannochloropsis. This study examined three PTMs: lysine acetylation (Kac), lysine β-hydroxybutyrylation (Kbhb), and phosphorylation. Using LC-MS/MS, we identified 4571 Kac sites, 7812 Kbhb sites, and 6237 phosphorylation sites across 2455, 3109, and 2786 proteins, respectively. Subcellular localization analysis revealed significant overlaps between Kac and Kbhb proteins, primarily in the chloroplast, cytosol, and nucleus, while phosphorylated proteins were predominantly located in the nucleus and chloroplast. Motif analysis highlighted specific amino acid enrichments around modification sites, with several motifs conserved. Additionally, 529 proteins harbored all three PTMs, underscoring the potential regulatory interplay. Kac, Kbhb, and phosphorylated proteins were particularly abundant in glycolysis, the TCA cycle, carbon fixation, and lipid metabolism pathways, influencing energy production and lipid accumulation. Based on previous transcriptome data under nutrient-limited conditions, these frequently modified key enzymes appear to be vital components in the response to abiotic stress. The presence of histone modifications related to Kac and Kbhb might also point to the epigenetic regulation in gene expression and stress adaptation. This comprehensive PTM landscape in N. oceanica provides a foundation of valuable insights into future metabolic engineering and biotechnological applications.

The Role of Biotechnology in Shaping the Future of Modern Agriculture

Biotechnology stands as a cornerstone in the evolution of modern agriculture, offering an array of innovative solutions poised to revolutionize crop productivity, enhance nutritional quality, and foster environmental sustainability. This comprehensive review delves into the multifaceted role of biotechnology in shaping the future landscape of agriculture, encompassing diverse realms such as genetic engineering, molecular breeding, and microbial biotechnology. Spanning from crop improvement to pest and disease management, soil health, and sustainable agriculture practices, the applications of biotechnology in agriculture are far-reaching and profound. Genetic engineering emerges as a powerful tool in the arsenal of agricultural biotechnology, facilitating the precise manipulation of plant genomes to confer desirable traits such as increased yield, enhanced nutritional content, and resistance to biotic and abiotic stresses. Molecular breeding techniques complement this approach, leveraging advances in genomics and bioinformatics to expedite the breeding process and develop high-performing crop varieties tailored to specific agroecological contexts, microbial biotechnology emerges as a pivotal force in sustainable agriculture, harnessing the power of beneficial microorganisms to improve soil fertility, enhance nutrient uptake, and suppress plant pathogens. From biofertilizers and biopesticides to bioremediation and phytoremediation, microbial-based solutions hold immense potential in mitigating environmental degradation and promoting agroecosystem resilience, the widespread adoption of biotechnology in agriculture is not devoid of societal, economic, and ethical considerations. Regarding food safety, environmental impact, and socio-economic equity underscore the need for rigorous regulatory frameworks and robust risk assessment protocols to ensure the responsible deployment of biotechnological innovations, public perceptions and fostering dialogue between stakeholders are essential for building trust and garnering support for biotechnology in agriculture. Education, transparency, and stakeholder engagement are paramount in navigating the complex terrain of biotechnology governance and fostering an enabling environment for innovation, the current state and future prospects of biotechnology in agriculture, this review endeavors to inform policymakers, researchers, and stakeholders about the transformative potential of biotechnology to address pressing global challenges such as food security, climate change, and environmental sustainability. Embracing the opportunities afforded by biotechnology holds the key to unlocking a more resilient, equitable, and sustainable agricultural future.

Advancing Chickpea Breeding: Omics Insights for Targeted Abiotic Stress Mitigation and Genetic Enhancement.

Chickpea is a major source of proteins and is considered the most economically vital food legume. Chickpea production is threatened by several abiotic and biotic factors worldwide. The main constraints limiting worldwide chickpea production are abiotic conditions such as drought, heat, salinity, and cold. It is clear that chickpea is treasured for its nutritive value, in particular its high protein content, and hence study of problems like drought, cold and salinity stresses are very important concerning chickpeas. In this regard, several physiological, biochemical, and molecular mechanisms are reviewed to confer tolerance to abiotic stress. The most crippling economic losses in agriculture occur due to these abiotic stressors, which affect plants in many ways. All these abiotic stresses affect the water relations of the plant, both at the cellular level as well as the whole-plant level, causing both specific and non-specific reactions, damage and adaptation reactions. These stresses share common features. Breeding programs use a huge collection of over 100,000 chickpea accessions as their foundation. Significant advancements in conventional breeding, including mutagenesis, gene/allele introgression, and germplasm introduction, have been made through this method. Abiotic tolerance and yield component selection are made easier by creating unique DNA markers for the genus Cicer, which has been made possible by developments in high-throughput sequencing and molecular biology. Transcriptomics, proteomics, and metabolomics have also made it possible to identify particular genes, proteins, and metabolites linked to chickpea tolerance to abiotic stress. Chickpea abiotic stress tolerance has been directly and potentially improved by biotechnological applications, which are covered by all 'Omics' approaches. It requires information on the abiotic stress response at the different molecular levels, which comprises gene expression analysis for metabolites or proteins and its impact on phenotype. Studies on chickpea genome-wide expression profiling have been conducted to determine important candidate genes and their regulatory networks for abiotic stress response. This study aimed to offer a detailed overview of the diverse 'Omics' approaches for resilience's to abiotic stresses on chickpea plants.

Flexible Non-Enzymatic Glucose Sensors: One-Step Green Synthesis of NiO Nanoporous Films via an Electro-Exploding Wire Technique.

In this study, we successfully synthesized nickel oxide (NiO) nanoparticles (NPs), i.e., samples NiO 24V, NiO 36V, and NiO 48V, via an environmentally friendly one-step electro-exploding wire technique by employing three distinct voltage levels of 24, 36, and 48 V, respectively. Sample NiO 48V showed the most rugged surface and smallest particle size, which helped to enhance electrocatalytic properties. The highest Ni3+ content of sample NiO 48V contributed to the increasing redox current and rendering highly enhanced chemical reactions and thereby improving their electrochemical properties and electrocatalytic performance in the glucose oxidation processes in alkaline (0.1 M NaOH, pH = 13) media. The NiO 48V electrode showcased an excellent linear detection range spanning from 0.1 to 1 mM, featuring a remarkable sensitivity of 1202 μA mM-1 cm-2 and an exceptionally low limit of detection (LOD) value of 0.25 μM. Remarkably, NiO NPs exhibited exceptional long-term stability, commendable reproducibility, favorable repeatability, and outstanding selectivity. This study also highlights the excellent operational performance of the NiO 48V electrode in real-world samples, such as commercially available beverages and human urine, highlighting the practical nature of these nonenzymatic sensors in real-life scenarios for the food industries, clinical diagnostics, and biotechnology applications.

Multi‐Color Optical Quasiparticle Laser Source Formed of a Pr3+ Doped Fiber Laser with a Dual‐Output Coupling Geometry

AbstractThe generation of multicolor (523, 605, 637, and 719 nm) optical quasiparticles (bimerons and skyrmions with topologically protected polarization textures) from a diode‐pumped Pr3+‐doped fluoro‐aluminate glass (Pr3+: WPFG) fiber simply with intra‐cavity plano‐convex lens and wedge‐plate and without any wavefront control elements, such as a spatial light modulator is demonstrated. This robust and cost‐saving system efficiently produces Bloch‐, Néel‐, and anti‐quasiparticles with high mode purity. In particular, the green optical quasiparticles will have the potential to explore many applications in materials science and biotechnologies.

Biotechnological and Biomedical Applications of Enzymes Involved in the Synthesis of Nucleosides and Nucleotides—2nd Edition

The study of nucleic acid derivatives and their role in cellular processes, first addressed in our previous Special Issue (https://www [...]

Single-molecule visualization of sequence-specific RNA binding by a designer PPR protein.

Pentatricopeptide repeat proteins (PPR) are a large family of modular RNA-binding proteins, whereby each module can be modified to bind to a specific ssRNA nucleobase. As such, there is interest in developing 'designer' PPRs (dPPRs) for a range of biotechnology applications, including diagnostics or in vivo localization of ssRNA species; however, the mechanistic details regarding how PPRs search for and bind to target sequences is unclear. To address this, we determined the structure of a dPPR bound to its target sequence and used two- and three-color single-molecule fluorescence resonance energy transfer to interrogate the mechanism of ssRNA binding to individual dPPRs in real time. We demonstrate that dPPRs are slower to bind longer ssRNA sequences (or could not bind at all) and that this is, in part, due to their propensity to form stable secondary structures that sequester the target sequence from dPPR. Importantly, dPPR binds only to its target sequence (i.e. it does not associate with non-target ssRNA sequences) and does not 'scan' longer ssRNA oligonucleotides for the target sequence. The kinetic constraints imposed by random 3D diffusion may explain the long-standing conundrum of why PPR proteins are abundant in organelles, but almost unknown outside them (i.e. in the cytosol and nucleus).

Efficient DNA-free co-targeting of nuclear genes in Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii, a model organism for unicellular green microalgae, is widely used in basic and applied research. Nonetheless, proceeding towards synthetic biology requires a full set of manipulation techniques for inserting, removing, or editing genes. Despite recent advancements in CRISPR/Cas9, still significant limitations in producing gene knock-outs are standing, including (i) unsatisfactory genome editing (GE) efficiency and (ii) uncontrolled DNA random insertion of antibiotic resistance markers. Thus, obtaining efficient gene targeting without using marker genes is instrumental in developing a pipeline for efficient engineering of strains for biotechnological applications. We developed an efficient DNA-free gene disruption strategy, relying on phenotypical identification of mutants, to (i) precisely determine its efficiency compared to marker-relying approaches and (ii) establish a new DNA-free editing tool. This study found that classical CRISPR Cas9-based GE for gene disruption in Chlamydomonas reinhardtii is mainly limited by DNA integration. With respect to previous results achieved on synchronized cell populations, we succeeded in increasing the GE efficiency of single gene targeting by about 200 times and up to 270 times by applying phosphate starvation. Moreover, we determined the efficiency of multiplex simultaneous gene disruption by using an additional gene target whose knock-out did not lead to a visible phenotype, achieving a co-targeting efficiency of 22%. These results expand the toolset of GE techniques and, additionally, lead the way to future strategies to generate complex genotypes or to functionally investigate gene families. Furthermore, the approach provides new perspectives on how GE can be applied to (non-) model microalgae species, targeting groups of candidate genes of high interest for basic research and biotechnological applications.

Numerical Computation and Statistical Interpretations of Heat Transfer of Tween-20/ethyl Acetate Nanofluid Flow with Melting Rheological Quality and Activation Energy

Tween-20 plays a significant role in the biological, food, and pharmaceutical industries. Additionally, it plays a vital role in improving the quality of reverse mechanisms of multi-drug resistance. This paper uses artificial neural computing to examine Tween-20 nanoparticles’ behaviors in a base fluid called ethyl acetate to enhance the applications in nanomedicine, drug delivery and biotechnology. The study focuses on the nonlinear model of a viscous fluid at the stagnation point, where mixed convection processes and activation energy are present. The study incorporates slip velocity and melting boundary conditions to examine heat and mass transfer, taking into account thermal and solutal stratification. Various fields of research and technology, specially in industrial engineering that include hydrodynamics, panto-graph systems, and biomedical mathematics, extensively utilize artificial computing. The datasets are based on velocity, temperature, and concentration outlines. The fourth-order Runge-Kutta method is used to generate the datasets. To validate the LMM-ABNNs, we have compared them with a numerical solution, showing a high level of agreement. We utilize the error histogram and mean square error results to validate the performance, scrutinize the training and testing methods, and explore the validity of the approximate answer. Furthermore, the quality characteristics such as skin friction, rate of heat, and mass movement (Nusselt and Sherwood numbers) are statistically analyzed to forecast the model’s durability. This article is the best example of investigating different fluid parameters with the latest artificial neural computing.

Synthetic translational coupling element for multiplexed signal processing and cellular control.

Repurposing natural systems to develop customized functions in biological systems is one of the main thrusts of synthetic biology. Translational coupling is a common phenomenon in diverse polycistronic operons for efficient allocation of limited genetic space and cellular resources. These beneficial features of translation coupling can provide exciting opportunities for creating novel synthetic biological devices. Here, we introduce a modular synthetic translational coupling element (synTCE) and integrate this design with de novo designed riboregulators,toehold switches. A systematic exploration of sequence domain variants for synTCEs led to the identification of critical design considerations for improving the system performance. Next, this design approach was seamlessly integrated into logic computations and applied to construct multi-output transcripts with well-defined stoichiometric control. This module was further applied to signaling cascades for combined signal transduction and multi-input/multi-output synthetic devices. Further, the synTCEs can precisely manipulate the N-terminal ends of output proteins, facilitating effective protein localization and cellular population control. Therefore, the synTCEs could enhance computational capability and applicability of riboregulators for reprogramming biological systems, leading to future applications in synthetic biology, metabolic engineering and biotechnology.

The Kinetics of Carbon-Carbon-Bond Formation in Metazoan Fatty Acid Synthase and its Impact on Product Fidelity.

Fatty acid synthase (FAS) multienzymes are responsible for de novo fatty acid biosynthesis and crucial in primary metabolism. Despite extensive research, the molecular details of the FAS catalytic mechanisms are still poorly understood. For example, the b-ketoacyl synthase (KS) catalyzes the fatty acid elongating carbon-carbon-bond formation, which is the key catalytic step in biosynthesis, but factors that determine the speed and accuracy of his reaction are still unclear. Here, we report enzyme kinetics of the KS-mediated carbon-carbon bond formation, enabled by a continuous fluorometric activity assay. We observe that the KS is likely rate-limiting to the fatty acid biosynthesis, its kinetics are adapted to the length of the bound fatty acyl chain, and that the KS is also responsible for the fidelity of biosynthesis by preventing intermediates from undergoing KS-mediated elongation. To provide mechanistic insight into KS selectivity, we performed computational molecular dynamics (MD) simulations. We identify positive cooperativity of the KS dimer, which we suggest to affect the conformational variability of the multienzyme. Advancing our knowledge about the KS molecular mechanism will pave the ground for engineering FAS for biotechnology applications and the design of new therapeutics targeting the fatty acid metabolism.

Argonaute-Based Nucleic Acid Detection Technology: Advantages, Current Status, Challenges, and Perspectives.

Rapid and accurate detection is a prerequisite for precise clinical diagnostics, ensuring food safety, and facilitating biotechnological applications. The Argonaute system, as a cutting-edge technique, has been successfully repurposed in biosensing beyond the CRISPR/Cas system (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins), which has been extensively researched, but recognition of PAM sequences remains restricted. Argonaute, as a programmable and target-activated nuclease, is repurposed for fabricating novel detection methods due to its unparalleled biological features. In this comprehensive review, we initially elaborate on the current methods for nucleic acid testing and programmable nucleases, followed by delving into the structure and nuclease activity of the Argonaute system. The advantages of Argonaute compared with the CRISPR/Cas system in nucleic acid detection are highlighted and discussed. Furthermore, we summarize the applications of Argonaute-based nucleic acid detection and provide an in-depth analysis of future perspectives and challenges. Recent research has demonstrated that Argonaute-based biosensing is an innovative and rapidly advancing technology that can overcome the limitations of existing methods and potentially replace them. In summary, the implementation of Argonaute and its integration with other technologies hold promise in developing customized and intelligent detection methods for nucleic acid testing across various aspects.

From coffee waste to nutritional gold: bioreactor cultivation of single‐cell protein from Candida sorboxylosa

AbstractBACKGROUNDIndustrial effluents are continuously discharged into the environment. These wastewaters contain valuable compounds that can be reused for biotechnological applications. Coffee wastewater (CWW) is a powerful effluent that can be used for single‐cell protein (SCP) production reaching important content (up to 80%). Several yeast species can be used for SCP production, but Candida species are commonly applied for this purpose (17 species reported including the novel C. sorboxylosa). In addition, SCP can be produced in bioreactors under controlled conditions under three operation modes. Thus, batch mode is frequently used but continuous mode presents interesting advantages in economic terms, although it has been poorly applied in SCP production.RESULTSThe initial evaluation under batch operation mode showed that volumetric oxygen transfer coefficient (kLa of 101 h−1) improved biomass production (1.39 g L−1) and SCP yield (59.9%) in C. sorboxylosa. Thus, continuous mode was established at selected kLa and feeding with optimized medium composed of 87.5% (v/v) CWW, 1.38 g L−1 yeast extract, and 7.24 g L−1 (NH4)2SO4, in order to provided necessary nutrients. In this sense, the process presented higher values in dry cell weight and SCP productivity (0.57 and 0.29 g L−1·h, respectively), achieving a 3.35‐ and 2.90‐fold increase in biomass and protein productivity, respectively, compared to batch mode. The SCP from C. sorboxylosa exhibited an interesting essential amino acid profile under continuous mode (33.704%).CONCLUSIONThe bioprocess highlights several advantages during bioreactor cultivation, including: (i) reduced energy consumption for temperature control; (ii) successful establishment of an initial continuous operation mode with promising performance; and (iii) SCP from C. sorboxylosa exhibited a notable composition of essential amino acids, which could be beneficial for potential use in animal feed. © 2024 Society of Chemical Industry (SCI).