Microbial Protein Research Articles

Evaluating bioproducts production in a purple phototrophic biofilm photobioreactor: Fuel-synthesis wastewater vs. simple substrates

This study evaluated the influence of different carbon sources on purple non-sulphur bacteria (PNSB) biofilm formation, and the production of polyhydroxybutyrate (PHB) and other value-added bio-products. Higher PNSB growth and biofilm formation were observed in fuel-synthesis wastewater (FSW), followed by acetate. Both FSW and acetate cultures contained a large proportion of Rhizobiales, while sugar substrates led to yeast enrichment. Protein contents reached 40–41 % for acetate, which is suitable for single cell protein (SCP) use. The maximum PHB content from the suspended and biofilm growth was obtained from glucose (18.1 ± 0.10 %, 12.9 ± 1.11 %), followed by FSW (14.9 ± 0.25 %, 11.2 ± 0.50 %). FSW showed the highest overall productivity for both protein and PHB. Pigment concentrations were low for all substrates. The study underscores substrate influence on microbial community and bioproducts potential. It highlights FSW as a promising substrate for PNSB, suggesting its treatment integration for resource recovery.

Microbial production of food compounds with carbon dioxide and derived low-carbon molecules as substrates

The construction and optimization of microbial cell factories are crucial steps and key technologies in achieving green biomanufacturing. As concern has been aroused regarding the excessive carbon dioxide (CO2) emissions and food security, a new and promising research field, microbial conversion of CO2 into food compounds, has emerged. The research in this field not only holds significant implications for achieving the carbon peaking and carbon neutrality goals but also plays a role in maintaining food security. This paper provides a comprehensive review and outlook of the research on utilizing CO2 and its derived low-carbon chemicals for the production of food compounds, focusing on the production of glucose, sugar derivatives, and single-cell proteins and the development of artificial CO2 fixation pathways.

The Influence of High-Concentrate Diet Supplemented with Tannin on Growth Performance, Rumen Fermentation, and Antioxidant Ability of Fattening Lambs.

This experiment aimed to study the effects of tannin supplementation on growth performance, rumen fermentation characteristics, apparent digestibility and serum biochemistry, and antioxidant and immune indexes in fattening lambs. A total of 36 male Hu sheep lambs (body weight = 15.83 ± 0.48 kg and days of age = 55 ± 2 d) were fed a high-concentrate diet and randomly divided into one of three groups of 12 animals each: control with no tannin (CON) and tannin treatments (TA1, 3 g/d per lamb; TA2, 6 g/d per lamb). The feeding experiment lasted for 60 d. The results showed that the average daily gain and ruminal propionate content of lambs in the TA1 group were higher (p < 0.05) than those in the CON group. Lambs fed tannin had significantly increased (p < 0.05) microbial protein and decreased (p < 0.05) ammonia nitrogen concentrations in the rumen. In addition, the crude protein and neutral detergent fiber digestibility of the TA2 group were significantly decreased (p < 0.05) as compared with the TA1 and CON groups, respectively. The serum concentrations of triglyceride, immunoglobulin A, and catalase and the total antioxidant capacity were higher (p < 0.05) in the TA1 group that those in the CON group, whereas an opposite trend of urea nitrogen, interleukin-1β, and malondialdehyde was found between the two groups. Also, tannin supplementation increased (p < 0.05) Lactobacillus and decreased (p < 0.05) Salmonella counts in the feces of lambs. Taken together, tannin supplementation can improve the growth performance, immunity, and antioxidant ability of fattening lambs fed a high-concentrate diet.

Potential Valorization of Banana Production Waste in Developing Countries: Bio-Engineering Aspects

Plant food production generates a lot of by-products (BPs). These BPs are majorly discarded into the environment, polluting it, or into landfills where they just decompose, providing no benefit and taking up storage space, causing financial costs. These plant BPs are biodegradable, but reusing them may provide a better outcome and profit. The vast majority of plant-based food BPs are polysaccharide polymers like gums, lignin, cellulose, and their derivatives. It is possible to utilize plant food production waste, like banana peels, leaves, pseudostems, and inflorescences, to produce bioethanol, single-cell protein, cellulase, citric acid, lactic acid, amylase, cosmetics, fodder additives, fertilizers, biodegradable fibers, sanitary pads, bio-films, pulp and paper, natural fiber-based composites, bio-sorbents, bio-plastic, and bio-electricity in the agro-industry, pharmaceutical, bio-medical, and bio-engineering fields. Moreover, the use of banana BPs seems to be a way of dealing with many issues in underdeveloped countries, providing a clean and ecological solution. The suggested idea might not only reduce the use of plastic but also mitigate waste pollution.

Production of Single Cell Protein (SCP) from the Peel Waste of Pea, Potato, and Banana by Aspergillus Flavus NRRL 21882 as an Efficient Organic Poultry Supplement.

Food protein deficit has become a major issue worldwide, particularly in underdeveloped countries. Scientists are searching for a variety of less expensive solutions to this issue. One of these less expensive methods is to create single cell protein as a substrate from leftover fruit and vegetable waste, which is typically thrown away. In this regard, the fungal strain Aspergillus flavus (NRRL 21882) was used for the synthesis of SCP (single cell protein) from the waste of banana, potato, and pea. In this manner, 30 samples were collected from the whole substrate with a share of 10 samples each from banana, potato, and pea peels, which were in turn dried and powdered finely. The fermentation process was done by the process of solid state fermentation. Aspergillus flavus (NRRL 21882) generated the highest percentage, i.e. 60.67%, of crude protein from the pea peels. The composition of amino acids in crude proteins was also investigated. The findings demonstrated that the highest percentage of aspartic acid (13.34 ± 0.80%) and glutamic acid (14.92 ± 0.69%) was found in A. flavus single cell protein produced from pea peels. Soybean was supplemented with single cell protein in the boilers' diet. Compared to all treated groups, there was a substantial (p ≤ 0.05) increase in the level of antibody titer against the Newcastle disease vaccine. The supplementation of single cell protein with soybean meal had no effect on the levels of liver enzymes. The liver enzymes found in all four groups (A, B, C, and D) were within normal limits. None of the examined groups experienced any change in the feed conversion ratio, with all groups exhibiting an average FCR of 1.6. The current study concludes that broiler health and immunity is increased by supplementing poultry feed with single cell protein.

Polyploid cancer cells reveal signatures of chemotherapy resistance.

Therapeutic resistance in cancer significantly contributes to mortality, with many patients eventually experiencing recurrence after initial treatment responses. Recent studies have identified therapy-resistant large polyploid cancer cells in patient tissues, particularly in late-stage prostate cancer, linking them to advanced disease and relapse. Here, we analyzed bone marrow aspirates from 44 advanced prostate cancer patients and found the presence of circulating tumor cells with increased genomic content (CTC-IGC) was significantly associated with poorer progression-free survival. Single cell copy number profiling of CTC-IGC displayed clonal origins with typical CTCs, suggesting complete polyploidization. Induced polyploid cancer cells from PC3 and MDA-MB-231 cell lines treated with docetaxel or cisplatin were examined through single cell DNA sequencing, RNA sequencing, and protein immunofluorescence. Novel RNA and protein markers, including HOMER1, TNFRSF9, and LRP1, were identified as linked to chemotherapy resistance. These markers were also present in a subset of patient CTCs and associated with recurrence in public gene expression data. This study highlights the prognostic significance of large polyploid tumor cells, their role in chemotherapy resistance, and their expression of markers tied to cancer relapse, offering new potential avenues for therapeutic development.

Resource recovery and treatment of wastewaters using filamentous fungi

Industrial wastewater, often characterized by its proximity to neutral pH, presents a promising opportunity for fungal utilization despite the prevalent preference of fungi for acidic conditions. This review addresses this discrepancy, highlighting the potential of certain industrial wastewaters, particularly those with low pH levels, for fungal biorefinery. Additionally, the economic implications of biomass recovery and compound separation, factors that require explicit were emphasized. Through an in-depth analysis of various industrial sectors, including food processing, textiles, pharmaceuticals, and paper-pulp, this study explores how filamentous fungi can effectively harness the nutrient-rich content of wastewaters to produce valuable resources. The pivotal role of ligninolytic enzymes synthesized by fungi in wastewater purification is examined, as well as their ability to absorb metal contaminants. Furthermore, the diverse benefits of fungal biorefinery are underscored, including the production of protein-rich single-cell protein, biolipids, enzymes, and organic acids, which not only enhance environmental sustainability but also foster economic growth. Finally, the challenges associated with scaling up fungal biorefinery processes for wastewater treatment are critically evaluated, providing valuable insights for future research and industrial implementation. This comprehensive analysis aims to elucidate the potential of fungal biorefinery in addressing industrial wastewater challenges while promoting sustainable resource utilization.

Recent Developments in the Application of Filamentous Fungus Aspergillus oryzae in Ruminant Feed.

The resource-intensive nature of the ruminant farming sector, which has been exacerbated by population growth and increasing pressure to reduce feed antibiotics and growth promoters, has sparked interest in looking for sustainable alternative feed sources to enhance ruminant production efficiency. Edible filamentous fungi, rich in macronutrients like proteins, offer promise in reducing the reliance on conventional protein sources and antimicrobials to improve feed quality and animal performance. The inclusion of single-cell proteins, particularly filamentous fungi, in ruminant feed has long been of scientific and industrial interest. This review focuses on the potential application of the extensively studied Aspergillus oryzae and its fermentation extracts in ruminant nutrition. It provides an overview of conventional ruminant feed ingredients, supplements, and efficiency. Additionally, this review analyzes the re-utilization of organic residues for A. oryzae cultivation and examines the effects of adding fungal extracts to ruminant feed on ruminal digestibility and animal performance, all within a circular bioeconomy framework.

Assessing a single-cell multi-omic analytic platform to characterize ex vivo-engineered T-cell therapy products.

Genetically engineered CD8+ T cells are being explored for the treatment of various cancers. Analytical characterization represents a major challenge in the development of genetically engineered cell therapies, especially assessing the potential off-target editing and product heterogeneity. As conventional sequencing techniques only provide information at the bulk level, they are unable to detect off-target CRISPR translocation or editing events occurring in minor cell subpopulations. In this study, we report the analytical development of a single-cell multi-omics DNA and protein assay to characterize genetically engineered cell products for safety and genotoxicity assessment. We were able to quantify on-target edits, off-target events, and potential translocations at the targeting loci with per-cell granularity, providing important characterization data of the final cell product. Conclusion: A single-cell multi-omics approach provides the resolution required to understand the composition of cellular products and identify critical quality attributes (CQAs).

Studies on chlorophenyl, thiophenyl, and biphenyl clubbed tetrahydro bipyrazole carbaldehydes: Synthesis, antimicrobial and antioxidant activity, SAR study, in-silico pharmacokinetics, toxicity, and molecular modeling

A library of chlorophenyl, thiophenyl, and biphenyl clubbed 3′,4,4′,5-tetrahydro-2′H-[1,3′-bipyrazole]-2′-carbaldehydes was produced via aldolic condensation followed by hetero-cyclization. Synthesized moieties were characterized by IR, 1H, and 13CNMR spectral analysis. The molecular docking was conducted utilizing microbial target proteins, 1JX9, and 5C1P to explore the binding interactions of scaffolds. Analogues 4f, 4l, 4r, and 4x exhibited the best docking score of -10.3 kcal/mol with 1JX9 whereas scaffolds 4l and 4o showed maximum docking of -10.4 and -10.2 kcal/mol with target protein 5C1P. Further, scaffolds 4f, 4I, 4l, 4o, 4 u, and 4v showed excellent (pMIC: 1.92 µM/mL) antibacterial activity against S. aureus, B. subtilis, E. coli, P. aeruginosa whereas; analogues 4c, 4l, 4o, 4r, and 4v exhibited admirable (pMIC: 1.92 µM/mL) antifungal activity against R. oryzae fungal strain. Also, analogues were tested for their radical scavenging ability and 4a, 4 g, 4i, 4p, and 4x exhibited good antioxidant activity with 4i standing out as a propitious scaffold with 93.25 % radical scavenging activity. SAR study revealed that biphenyl, 4‑hydroxy-3‑methoxy phenyl, and thiophenyl moieties on the bipyrazole-carbaldehyde scaffold are accountable for virtuous antibacterial, antifungal, and antioxidant activity, respectively. Further in-silico ADME, pharmacokinetic, and toxicity properties were studied, forecasting less-toxic analogues (300 < LD50 ≤ 5000). These findings collectively underscore the potential of the synthesized compounds, with 4l, 4o, 4v and 4x standing out as particularly promising candidates for further exploration.

Conditional Optimization of Single Cell Protein Production from Crude Oil by &amp;lt;i&amp;gt;Pseudomonas aeruginosa&amp;lt;/i&amp;gt;

The study was carried out to investigate the possible biodegradation of crude oil as a carbon by the bacterium &amp;lt;i&amp;gt;Pseudomonas aeruginosa&amp;lt;/i&amp;gt; isolated from marine environment (Ras El-Menkar- Benghazi- Libya) using basal yeast extract protease peptone-3 (BYP) enriched medium. The isolated bacterium was identified and characterized according to its cultural condition and microbial biochemical properties. Different experiments were developed throughout this study to stimulate bacterial growth and production of single cell protein (SCP). The results show that the optimal concentration of crude oil as a carbon source for the highest bacterial growth (1.14g/l), and production of SCP (0.65g/l; 57.02% of the biomass dry weight) was 1%. This was required to utilize up to 50.6% of oil as a carbon source. As to the nitrogen source, the optimal concentration of ammonium chloride was 0.1%, in which the bacterial growth and SCP production increased to 1.23 g/l and 0.67 g/l respectively. The stimulating effects of organic and inorganic factors on the bacterial growth and SCP production was also tested. Addition of inorganic nutrients such as potassium phosphate (0.05%), magnesium sulphate (0.01%), and organic nutrient in the form of yeast extract (0.1%) to the fermentation medium slightly promoted the bacterial growth which reflected positively on SCP production and the percentage of the consumed crude oil, (&amp;gt;57%) at final pH value of 8.0. The obtained results indicated that the isolated&amp;lt;i&amp;gt; Pseudomonas aeruginosa&amp;lt;/i&amp;gt; posses the ability to utilize the crude oil and use it as a carbon for bacterial growth and production of SCP.

SANS investigation of fungal loosenins reveal substrate dependent impacts of protein 1 action on inter-fibril distance and packing order of cellulosic substrates.

Microbial expansin-related proteins include fungal loosenins, which have been previously shown to disrupt cellulose networks and enhance the enzymatic conversion of cellulosic substrates. Despite showing beneficial impacts to cellulose processing, detailed characterization of cellulosic materials after loosenin treatment is lacking. In this study, small-angle neutron scattering (SANS) was used to investigate the effects of three recombinantly produced loosenins that originate from Phanerochaete carnosa, PcaLOOL7, PcaLOOL9, and PcaLOOL12, on the organization of holocellulose preparations from Eucalyptus and Spruce wood samples. Whereas the SANS analysis of Spruce holocellulose revealed an increase in interfibril spacing of neighboring cellulose microfibrils following treatment with PcaLOOL12 and to a lesser extent PcaLOOL7, the analysis of Eucalyptus holocellulose revealed a reduction in packing number following treatment with PcaLOOL12 and to a lesser extent PcaLOOL9. Parallel SEC-SAXS characterization of PcaLOOL7, PcaLOOL9, and PcaLOOL12 indicated the proteins likely function as monomers; moreover, all appear to retain a flexible disordered N-terminus and folded C-terminal region. The comparatively high impact of PcaLOOL12 motivated its NMR structural characterization, revealing a double-psi b-barrel (DPBB) domain surrounded by three alpha-helices - the largest nestled against the DPBB core and the other two part of loops extending from the core. The SANS analysis of PcaLOOL action on holocellulose samples confirms their ability to disrupt cellulose fiber networks and suggests a progression from reducing microfibril packing to increasing interfibril distance. The most impactful PcaLOOL, PcaLOOL12, was previously observed to be the most highly expressed loosenin in P. carnosa. Its structural characterization herein reveals its stabilization through two disulfide linkages, and an extended N-terminal region distal to a negatively charged and surface accessible polysaccharide binding groove.

Influence of Feed and Feeding Strategies on Rumen Microbiome: A Review

The presence of complex microorganisms in the rumen of ruminants are able to process the lignocellulose from low quality feed and fodder into volatile fatty acids, convert non-protein nitrogen into microbial protein and synthesize B vitamins, thus, ruminants are able to produce food of animal origin without competition for feed with non-ruminants and man. On the other hand, they contribute 14 per cent of anthropogenic methane, released annually into the atmosphere. Manipulating the rumen microbiome is considered as an important approach to reduce environmental impact, while increasing production efficiency in ruminants. A systematic literature search was undertaken and relevant research and review articles were downloaded, classified and salient points extracted to prepare the narrative review on “Influence of Feed and Feeding strategies on Rumen microbiome”. The microbial community in the rumen is the most diverse composed of bacteria, archaea, protozoa, fungi and uncharacterized virome. Differences in ruminal microbial community compositions are predominantly attributable to diet. Dietary manipulation offers a viable solution to alter the rumen microbiome so that the present-day challenges faced by the ruminant livestock industry is addressed.

MRGM: An enhanced catalog of mouse gut microbial genomes substantially broadening taxonomic and functional landscapes.

Mouse gut microbiome research is pivotal for understanding the human gut microbiome, providing insights into disease modeling, host-microbe interactions, and the dietary influence on the gut microbiome. To enhance the translational value of mouse gut microbiome studies, we need detailed and high-quality catalogs of mouse gut microbial genomes. We introduce the Mouse Reference Gut Microbiome (MRGM), a comprehensive catalog with 42,245 non-redundant mouse gut bacterial genomes across 1,524 species. MRGM marks a 40% increase in the known taxonomic diversity of mouse gut microbes, capturing previously underrepresented lineages through refined genome quality assessment techniques. MRGM not only broadens the taxonomic landscape but also enriches the functional landscape of the mouse gut microbiome. Using deep learning, we have elevated the Gene Ontology annotation rate for mouse gut microbial proteins from 3.2% with orthology to 60%, marking an over 18-fold increase. MRGM supports both DNA- and marker-based taxonomic profiling by providing custom databases, surpassing previous catalogs in performance. Finally, taxonomic and functional comparisons between human and mouse gut microbiota reveal diet-driven divergences in their taxonomic composition and functional enrichment. Overall, our study highlights the value of high-quality microbial genome catalogs in advancing our understanding of the co-evolution between gut microbes and their host.

Assessing the efficacy of date-pits holocellulose as a novel additive candidate for ruminant feeding.

Holocellulose (HC) fraction extracted from date-pits was evaluated as a novel feed additive for ruminant feeding. This study was performed to investigate the effectiveness of the HC additive on rumen fermentation, methane (CH4) production, and diet degradability over 24h of in vitro incubation. Three independent incubation trials were conducted over three consecutive weeks, employing the same in vitro methodology to assess four treatment doses in a completely randomized design. The experimental diet incorporated four increasing doses of HC, containing HC at 0 (HC0), 10 (HC10), 20 (HC20), and 30 (HC30) g/kg dry matter (DM). In vitro gas production (GP) and CH4 production, volatile fatty acids (VFAs) concentration, protozoa accounts, degraded organic matter (DOM), metabolizable and net energy (ME and NE), and hydrogen (H2) estimates were measured. No significant differences in ruminal pH were observed as the HC doses gradually increased. All incremental doses of HC additive over 24h resulted in a linear increase in GP (P < 0.001), DOM (P < 0.001), total VFAs (P = 0.011), and propionate (P < 0.001) concentrations, as well as estimated energy (ME and NE) (P < 0.05) and microbial protein (P = 0.017) values. However, the inclusion of increasing doses of HC in the diet displayed linear reductions in the net CH4 production (ml/kg DOM; P = 0.002), protozoa abundance (P = 0.027); acetate (P = 0.029), and butyrate (P < 0.001) concentrations, the acetate-to-propionate ratio (P < 0.001), and the estimated net H2 production concentration (P = 0.049). Thus, the use of date-pits HC additive generated positive ruminal fermentability, including increased total VFAs and a reduction in the acetate-to-propionate ratio, leading to decreased CH4 output over 24h of in vitro incubation. Hence, HC could be considered a potent feed additive (at up to 30g/kg DM), demonstrating promising CH4-mitigating competency and thereby enhancing energy-use efficiency in ruminants.

Increasing fish production in recirculating aquaculture system by integrating a biofloc-worm reactor for protein recovery

Aquaculture, producing half of global fish production, offers a high-quality protein source for humans. Improving nitrogen use efficiency (NUE) through microbial protein recovery is crucial for increasing fish production and reducing environmental footprint. However, the poor palatability and high moisture content of microbial protein make its utilization challenging. Here, a biofloc-worm reactor was integrated into a recirculating aquaculture system (BW_RAS) for the first time to convert microbial protein into Tubificidae (Oligochaeta) biomass, which was used as direct feed for culturing fish. Batch experiments indicated that an aeration rate of 0.132 m3L−1h−1 and a worm density of 0.3 g cm−2 on the carrier were optimal for microbial biomass growth and worm predation, respectively. Compared to the biofloc reactor-based recirculating aquaculture system (B_RAS), the BW_RAS improved water quality, NUE, and fish production by 17.1 % during a 120-day aquaculture period. The abundance of heterotrophic aerobic denitrifier Deinococcus in BW_RAS was one order of magnitude higher than in B_RAS, while heterotrophic bacteria Mycobacterium was more abundant in B_RAS. Denitrifiers cooperated with organic matter degraders and nitrogen assimilation bacteria for protein recovery and gaseous nitrogen loss while competing with predatory bacteria. Function prediction and qPCR indicated greater aerobic respiration, nitrate assimilation, nitrification (AOB-amoA), and denitrification (napA, nirK, nirS, nosZI), but lower fermentation in BWR compared to BR. This study demonstrated that BW_RAS increased microbial protein production and aerobic nitrogen cycling through ongoing worm predation, further enhancing fish production to a commercially viable level.

AlphaFold 2-based stacking model for protein solubility prediction and its transferability on seed storage proteins

Accurate protein solubility prediction is crucial in screening suitable candidates for food application. Existing models often rely only on sequences, overlooking important structural details. In this study, a regression model for protein solubility was developed using both the sequences and predicted structures of 2983 E. coli proteins. The sequence and structural level properties of the proteins were bioinformatically extracted and subjected to multilayer perceptron (MLP). Moreover, residue level features and contact maps were utilized to construct a graph convolutional network (GCN). The out-of-fold predictions of the two models were combined and fed into multiple meta-regressors to create a stacking model. The stacking model with support vector regressor (SVR) achieved R2 of 0.502 and 0.468 on test and external validation datasets, respectively, displaying higher performance compared to existing regression models. Based on the improved performance compared to its based models, the stacking model effectively captured the strength of its base models as well as the significance of the different features used. Furthermore, the model's transferability was indirectly validated on a dataset of seed storage proteins using Osborne definition as well as on a case study using molecular dynamic simulation, showing potential for application beyond microbial proteins to food and agriculture-related ones.

Severity of Rolling Reconstituted High Moisture Barley on Ensiling Characteristics and In Vitro Ruminal Fermentation

Barley grain sources with variable kernel sizes makes adequate and consistent processing of kernels challenging. This study evaluated how the severity of processing for reconstituted high moisture (65% on DM basis) barley (RHB) affects ensiling characteristics and in vitro ruminal fermentation. Three independent sources of light (<630 g/500 mL) and heavy (>670 g/500 mL) barley were blended to create 4 sources of variable kernel sized barley (646 g/500 mL). Reconstituted barley was rolled through a roller gap width of 1.40 (RHBF), 1.86 (RHBM), or 2.31 mm (RHBC) and ensiled for 1 or 5 mo with dry rolled barley (DRB; roller gap width 1.86 mm) used as a control. The 1-mo RHB and the DRB were further evaluated using the artificial rumen technique (RUSITEC) to investigate the effects of severity of processing for RHB on ruminal fermentation, and gas, methane, and microbial protein production. Using a randomized complete block design (n = 4), 16 fermenters from 2 RUSITEC apparatuses were used to assess the 4 sources and 4 processing treatments. The addition of water increased kernel width before rolling and resulted in increased kernel length, width, and thickness for RHB relative to dry rolled barley. Increasing processing severity for RHB linearly increased kernel width. The percentage of fine particles (<1.18 mm) was greater for DRB than RHBF, but did not differ by processing severity for RHB. Dry matter, organic matter, and starch disappearance were not different between DRB and RHBF, but linearly increased with increasing processing severity for RHB. Fermenter pH tended to be less for DRB relative to RHBF. In conclusion, the reduction in fine particles with the addition of water for RHB may prevent a decline in fermenter pH and when processed to achieve the same PI using a smaller roller gap width, yielded similar DM and OM disappearance suggesting a lesser risk for low ruminal pH without compromising digestibility.

Enhancing Protein Content in Wild-Type Saccharomyces cerevisiae via Random Mutagenesis and Optimized Fermentation Conditions.

Single-cell protein (SCP) derived from microorganisms is widely recognized as a viable alternative protein source for the future. Nevertheless, the commercialization of yeast-based SCP is hampered by its relatively low protein content. Therefore, this study aimed to enhance the protein content of Saccharomyces cerevisiae via random mutagenesis. To achieve this, S. cerevisiae KCCM 51811, which exhibited the highest protein concentration among 20 edible S. cerevisiae strains, was selected as a chassis strain. Subsequently, a KCCM 51811 mutant library was constructed (through UV irradiation) and screened to isolate mutants exhibiting high protein content and/or concentration. Among the 174 mutant strains studied, the #126 mutant exhibited a remarkable 43% and 36% higher protein content and concentration, respectively, compared to the parental strain. Finally, the #126 mutant was cultured in a fed-batch system using molasses and corn-steep liquor, resulting in a protein concentration of 21.6 g/l in 100 h, which was 18% higher than that produced by the parental strain. These findings underscore the potential of our approach for the cost-effective production of food-grade SCP.

Valorisation of agricultural residues into Thauera sp. Sel9 microbial proteins for aquaculture

Unconventional protein sources are necessary to tackle the increasing demand for food. Microbial proteins (MP) are an alternative source of proteins for feed or food, suitable as feed for aquaculture. Substituting fishmeal with MP obtained from agricultural wastes could reduce the environmental burden of aquaculture and help with waste management. In this study, pure culture MP from the PHA-producer Thauera sp. Sel9 were obtained from agricultural residues (agro-zootechnical digestate and pasta industry leftovers). The produced MP was used in feeding tests with the model fish zebrafish (Danio rerio) to assess potential toxic effects and evaluate overall fish health. The obtained MP was rich in protein (59.5 % w/w over TS) and PHAs (15.0 %) and comprised all fish essential amino acids. The chemical scores and essential amino acid index confirmed the excellent quality of the MP. The feeding tests with 50 % feed substitution with MP resulted in survival rates (80–88 %) comparable to the control group (78 %), with only 100 % MP showing increased mortality. Thauera MP obtained from agricultural residues has the potential to become a partial fishmeal substitute in fish-farming.