Producing Strain Research Articles

Discovery of nostatin A, an azole-containing proteusin with prominent cytostatic and pro-apoptotic activity.

Ribosomally synthesized and post-translationally modified peptides (RiPPs) are intriguing compounds with potential pharmacological applications. While many RiPPs are known as antimicrobial agents, a limited number of RiPPs with anti-proliferative effects in cancer cells are available. Here we report the discovery of nostatin A (NosA), a highly modified RiPP belonging among nitrile hydratase-like leader peptide RiPPs (proteusins), isolated from a terrestrial cyanobacterium Nostoc sp. Its structure was established based on the core peptide sequence encoded in the biosynthetic gene cluster recovered from the producing strain and subsequent detailed nuclear magnetic resonance and high-resolution mass spectrometry analyses. NosA, composed of a 30 amino-acid peptide core, features a unique combination of moieties previously not reported in RiPPs: the simultaneous presence of oxazole/thiazole heterocycles, dehydrobutyrine/dehydroalanine residues, and a sactionine bond. NosA includes an isobutyl-modified proline residue, highly unusual in natural products. NosA inhibits proliferation of multiple cancer cell lines at low nanomolar concentration while showing no hemolysis. It induces cell cycle arrest in S-phase followed by mitochondrial apoptosis employing a mechanism different from known tubulin binding and DNA damaging compounds. NosA also inhibits Staphylococcus strains while it exhibits no effect in other tested bacteria or yeasts. Due to its novel structure and selective bioactivity, NosA represents an excellent candidate for combinatorial chemistry approaches leading to development of novel NosA-based lead compounds.

In vivo and in vitro reconstitution of biosynthesis of N-prenylated phenazines revealing diverse phenazine-modifying enzymes.

Phenazine natural products play various roles such as signal molecules, antibiotics, or electron carriers in their producer strains. Among these products, phenazinomycin and lavanducyanin, which are produced by Streptomyces species, are characterized by an N-alkyl modification. Herein, we established the biosynthetic pathways for these two phenazine natural products. Gene-disruption experiments and in vitro reconstitution of the phenazine-tailoring pathway revealed the late steps of the biosynthetic pathway of the phenazines. The class II terpene cyclase homolog Pzm1 catalyzes the cyclization reaction of farnesyl diphosphate to form monocyclic farnesyl diphosphate. Additionally, the prenyltransferase homolog PzmP functions as the N-prenyltransferase of 5,10-dihydrophenazine-1-carboxylic acid. The flavin monooxygenase homolog PzmS catalyzes the oxidative decarboxylation of prenylated 5,10-dihydrophenazine-1-carboxylic acid to yield phenazinomycin. This study highlights unprecedented modification enzymes for phenazine natural products.

Interferon gamma kinetics associated with immunodiagnostic techniques in experimental goats infected with biofilm producer and non-biofilm producer strains of Corynebacterium pseudotuberculosis

Caseous lymphadenitis can be diagnosed using serological tests, mainly the enzyme-linked immunosorbent assay (ELISA). In this work, we evaluated interferon-gamma production in goats infected and not infected with Corynebacterium pseudotuberculosis, correlating it with the ELISA diagnostic test and analyzing the cellular and humoral responses, respectively. Eighteen Canindé goats experimentally infected with biofilm-producing and non-biofilm-producing strains were used. The sensitivity of the antigens was above 80%, with seroconversion observed from 14 days post-infection. A significant band between 64 and 70 kDa was shown in Western blotting. The production of IFN-y shows a tendency towards greater production in goats infected with the biofilm-producing strain stimulated by the TPP antigen of the same strain. The results help better understand the cellular and humoral response and antigenicity of the strains under study, correlated with the production of IFN-y, which plays a vital role in infection.

Harnessing Gram-negative bacteria for novel anti-Gram-negative antibiotics.

Natural products have proven themselves as a valuable resource for antibiotics. However, in view of increasing antimicrobial resistance, there is an urgent need for new, structurally diverse agents that have the potential to overcome resistance and treat Gram-negative pathogens in particular. Historically, the search for new antibiotics was strongly focussed on the very successful Actinobacteria. On the other hand, other producer strains have been under-sampled and their potential for the production of bioactive natural products has been underestimated. In this mini-review, we highlight prominent examples of novel anti-Gram negative natural products produced by Gram-negative bacteria that are currently in lead optimisation or preclinical development. Furthermore, we will provide insights into the considerations and strategies behind the discovery of these agents and their putative applications.

Efficient utilization of xylose requires CO2 fixation in Synechococcus elongatus PCC 7942

Cyanobacteria show great promise as autotrophic hosts for the renewable biosynthesis of useful chemicals from CO2 and light. While they can efficiently fix CO2, cyanobacteria are generally outperformed by heterotrophic production hosts in terms of productivity and titer. Photomixotrophy, or co-utilization of sugars and CO2 as carbon feedstocks, has been implemented in cyanobacteria to greatly improve productivity and titers of several chemical products. We introduced xylose photomixotrophy to a 2,3-butanediol producing strain of Synechococcus elongatus PCC 7942 and characterized the effect of gene knockouts, changing pathway expression levels, and changing growth conditions on chemical production. Interestingly, 2,3-butanediol production was almost completely inhibited in the absence of added CO2. Untargeted metabolomics implied that RuBisCO was a significant bottleneck, especially at ambient CO2 levels, restricting the supply of lower glycolysis metabolites needed for 2,3-butanediol production. The dependence of the strain on elevated CO2 levels suggests some practical limitations on how xylose photomixotrophy can be efficiently carried out in S. elongatus.

Poly-γ-glutamic production by solid-state fermentation of Bacillus natto in ammonia nitrogen movement and soil water retention processes

A high polyglutamic acid (γ-PGA) producing strain of Bacillus natto UV-40–50 was screened by ultraviolet mutagenesis treatment and identified as still belonging to the Bacillus specie. The optimal fermentation medium composition for solid state fermentation (SSF) of B. natto strain UV-40–50 strain was determined by one-way analysis of variance, under which the yield of γ-PGA was 55.19 g/kg, and the presence and molecular weight of γ-PGA in the γ-PGA-purified samples were determined by a series of characterizations. The purification ability of the unseparated solid fermentation product (SFP) on ammonia nitrogen and nitrite in the water column, as well as its effect on soil water retention, germination rate and seedling length of lettuce and cabbage were further investigated. The results showed that the addition of 1 g/m3 SFP could effectively remove more than 60 % of ammonia nitrogen and more than 40 % of nitrite in the water body; the addition of 0.01 % SFP could increase the water retention capacity of cabbage soil by 2.13 times, and increase the water retention capacity of lettuce soil by 12 %; at the same time, the SFP could also significantly increase the germination rate and seedling length of both cabbage and lettuce.

Halogenated Adenine and Adenosine Natural Products in Streptomyces sp. JCM9888

AbstractAscamycin 1 and dealanylascamycin 2 are adenosine containing nucleoside antibiotics carrying the rare 5’‐O‐sulfamoyl ribose motif and a chlorine atom attached to C2 of the adenine ring. Gene knockouts (ΔacmK and ΔacmJ) in the producing strain, Streptomyces sp. JCM9888, generated mutants that were disabled in the assembly of the sulfamoyl moiety. This resulted in the production of 2‐chloroadenosine 3 and 2‐chloroadenine 4, indicating that purine halogenation can occur independently of sulfamoylation. Incubations in media enriched in bromide content resulted also in the identification of 2‐bromoadenine 5, 2‐bromoadenosine 6 and 5′‐O‐sulfamoyl‐2‐bromoadenosine 7.

Production of bacteriocin-like inhibitory substance (BLIS) by Staphylococcus spp. isolates from dogs.

In the present study, 39 canine isolates of Staphylococcus spp. were tested for antimicrobial substance (AMS) production. Seven AMS producers were identified, whose products exhibited a non-acidic character and a proteinaceous nature, therefore being considered bacteriocin-like inhibitory substances (BLIS). The producer strains of BLIS P1, P16 and I3 showed a broad spectrum of antimicrobial activity. Human, veterinary and plant pathogens, such as Listeria monocytogenes, Klebsiella pneumoniae, Staphylococcus spp. and Clavibacter michiganensis, were among the inhibited micro-organisms, suggesting the potential biotechnological application of these peptides. MALDI-TOF mass spectrometry and 16S rDNA sequencing identified the producer strains of BLIS P1, P16 and I3 as Staphylococcus pseudintermedius P1, Staphylococcus schleiferi P16 and Staphylococcus pseudintermedius I3. The plasmid profile of these strains suggests that the BLIS production is linked to biosynthetic genes located on plasmids. PCR analyses revealed that BLIS P1, P16 and I3 are different from 11 staphylococcins already described in the literature and that their genomic DNAs do not carry the most prevalent staphylococcal enterotoxin genes. The highest levels of BLIS production were achieved after 18-24h of growth of the producer strains in TSB medium. Moreover, BLIS P1 and I3 exhibited high resistance to temperature and pH variations, and BLIS P16 maintained 100% of its activity in almost all conditions tested. The characteristics associated with BLIS P1, P16 and I3 described in this work encourage further investigation of these substances, in addition to this study being the first report of BLIS production by a strain of S. schleiferi.

Isolation, Screening and Identification of Biosurfactant Producing Strain Nocardiopsis dassonvillei var B2 From Oil Contaminated Soil.

Petroleum and other oil manufacturing industries contribute to environmental pollution by releasing hazardous hydrocarbons. Biosurfactants offer a sustainable solution for mitigating oil pollution through emulsification processes, safeguarding agricultural soils, aquatic ecosystems, and human health. This study focuses on isolating, screening, and identifying actinomycetes producing biosurfactant from oil-polluted soil in the naval dockyard of Visakhapatnam. Biosurfactant production was successfully achieved utilizing Kim's medium, which was supplemented with olive oil serving as the carbon source. The evaluation involved preliminary identification tests, including oil displacement, Parafilm-M, and lipase activity assays, using sodium lauryl sulfate as the standard reference. Surface tension and emulsification index measurements were conducted, and the chemical composition of glycolipids and phospholipids was elucidated using phenol-sulfuric acid and phosphate assays. Glycolipids, specifically identified as rhamnolipids, were confirmed via cetyltrimethylammonium bromide (CTAB) testing and quantitatively analyzed using the orcinol method. The cell-free broth exhibited antagonistic activity against Gram-positive and negative bacilli.16S rRNA sequencing-based phylogenetic analysis was carried out by the NCIM, Pune, with the gene sequence being deposited in GenBank. Further characterization of isolate B2 included scanning electron microscopy (SEM) analysis, as well as physiological and biochemical assays. This study highlights the ability of Nocardiopsis dassonvillei var. B2, isolated from oil-polluted soil, to produce biosurfactants, specifically glycolipids identified as rhamnolipids. Our findings represent the first reported instance of biosurfactant production from isolate B2 originating from the naval dockyard in Visakhapatnam, Andhra Pradesh, India.

The Molecules Gateway: A Homogeneous, Searchable Database of 150k Annotated Molecules from Actinomycetes.

Natural products are a sustainable resource for drug discovery, but their identification in complex mixtures remains a daunting task. We present an automated pipeline that compares, harmonizes and ranks the annotations of LC-HRMS data by different tools. When applied to 7,400 extracts derived from 6,566 strains belonging to 86 actinomycete genera, it yielded 150,000 molecules after processing over 50 million MS features. The web-based Molecules Gateway provides a highly interactive access to experimental and calculated data for these molecules, along with the metadata related to extracts and producer strains. We show how the Molecules Gateway can be used to rapidly identify known hard to find microbial products, unreported analogs of known families and not yet described metabolites. The Molecules Gateway, which complements available repositories, contains annotated MS data, both acquired and computationally processed under an identical workflow, making it suitable for global analyses which reveal a large and untapped chemical diversity afforded by actinomycetes.

Composition of biopesticides for crop protection – current state and prospects (review)

Aim. To summarise and analyse available information on research into biopesticide compositions aimed at enhancing their effectiveness and to assess prospects for the development of this field so as to determine the influence of components on the effectiveness of biopesticides for agricultural crop protection.This review article presents the results of studies of components included in the compositions of microorganism‐based biopesticides based on a system analysis. It is noted that the selection of biopesticide components must be carried out in accordance with the requirements for products recommended for use in ecological and organic farming systems. It is indicated that the choice of optimal formulation is associated with the characteristics of the producer strains, their trophic needs and resistance to biotic and abiotic factors and with the characteristics of the biopesticide application technology (seed treatment, application to the soil or treatment of plants during vegetation), as well as the type of equipment used.Knowledge of the basic requirements for biopesticide formulations will help maintain the viability and biological activity of microbial agents during storage of biopesticides, taking into account the sensitivity of producer strains to environmental conditions. Selection of optimal carriers, adhesives, stabilisers, thickeners and synergists, taking into account the technology of application of the biopesticide and the target pathogen, will facilitate the development and appearance on the market of new biopesticides for protecting agricultural crops from diseases.

Production of α-ketoisovalerate with whey powder by systemic metabolic engineering of Klebsiella oxytoca

BackgroundWhey, which has high biochemical oxygen demand and chemical oxygen demand, is mass-produced as a major by-product of the dairying industry. Microbial fermentation using whey as the carbon source may convert this potential pollutant into value-added products. This study investigated the potential of using whey powder to produce α-ketoisovalerate, an important platform chemical.ResultsKlebsiella oxytoca VKO-9, an efficient L-valine producing strain belonging to Risk Group 1 organism, was selected for the production of α-ketoisovalerate. The leucine dehydrogenase and branched-chain α-keto acid dehydrogenase, which catalyzed the reductive amination and oxidative decarboxylation of α-ketoisovalerate, respectively, were inactivated to enhance the accumulation of α-ketoisovalerate. The production of α-ketoisovalerate was also improved through overexpressing α-acetolactate synthase responsible for pyruvate polymerization and mutant acetohydroxyacid isomeroreductase related to α-acetolactate reduction. The obtained strain K. oxytoca KIV-7 produced 37.3 g/L of α-ketoisovalerate from lactose, the major utilizable carbohydrate in whey. In addition, K. oxytoca KIV-7 also produced α-ketoisovalerate from whey powder with a concentration of 40.7 g/L and a yield of 0.418 g/g.ConclusionThe process introduced in this study enabled efficient α-ketoisovalerate production from low-cost substrate whey powder. Since the key genes for α-ketoisovalerate generation were integrated in genome of K. oxytoca KIV-7 and constitutively expressed, this strain is promising in stable α-ketoisovalerate fermentation and can be used as a chassis strain for α-ketoisovalerate derivatives production.

Biosynthesis of antibacterial bacterial cellulose in-situ using yellow wine wastewater through co-fermentation

Bacterial cellulose (BC) is extensively used in many fields due to its excellent physical properties; however, its high production cost and low output remain a challenge. The production of conventional yellow wine typically results in the generation of a substantial quantity of rice soaking wastewater (RSW). In this study, BC producing strain was isolated and identified from Kombucha. BC was produced directly using unsterilized RSW as the medium. High-yield BC was obtained by co-fermentation of isolated Novacetimonas hansenii S-08 with natural fermentation flora from RSW. The culture conditions were optimized by combining RSW medium and seed liquid of HS medium at a ratio of 5:1, using unsterilized RSW as a medium containing 40 g/L fructose corn syrup and 2 % ethanol. The results of FTIR, XRD, TGA, SEM analysis, and mechanical properties showed that the physical properties of BC-RSW were comparable with ordinary BC. The BC-RSW had strong inhibitory effects on both Gram-positive and Gram-negative bacteria. In conclusion, the wastewater produced by yellow wine enterprises could be directly used as an alternative medium for the sustainable biosynthesis of antibacterial BC in situ.

High-efficiency breeding of Bacillus siamensis with hyper macrolactins production using physical mutagenesis and a high-throughput culture system

Macrolactins have attracted considerable attention due to their value and application in medicine and agriculture. However, poor yields severely hinder their broader application in these fields. This study aimed to improve macrolactins production in Bacillus siamensis using a combined atmospheric and room-temperature plasma mutagenesis and a microbial microdroplet culture system. After 25 days of treatment, a desirable strain with macrolactins production 3.0-fold higher than that of the parental strain was successfully selected. The addition of 30 mg/L ZnSO4 further increased macrolactins production to 503 ± 37.6 μg/mL, representing a 30.9 % improvement in production compared to controls. Based on transcriptome analysis, the synthesis pathways of amino acids, fengycin, and surfactin were found to be downregulated in IMD4036. Further fermentation experiments confirmed that inhibition of the comparative fengycin synthesis pathway was potentially driving the increased production of macrolactins. The strategies and possible mechanisms detailed in this study can provide insight into enhancing the production of other secondary metabolites toxic to the producer strains.

Screening of lactic acid bacteria for highly producing extra and intracellular folate and their potential use in production of bio-enriched yogurt

In this study, firstly, Microbial Assay (MA) was performed to evaluate the levels of intra and extracellular folate (Quatro folate) produced by 43 lactic acid bacteria strains isolated from traditional Iranian dairy products. The results obtained from strain screening showed that two strains of Lactobacillus delbrueckii subsp. bulgaricus, two strains of Streptococcus thermophilus, two strains of Lactobacillus delbrueckii subsp. lactis and one strain of Lactobacillus helveticus are highly folate producing. However, two strains of Lactobacillus delbrueckii subsp. bulgaricus produced higher amounts of folate (133.23 μg/mL and 102 μg/mL, respectively). In the second phase, eight yogurt samples were produced by using the two highest folate producing strains and compared with the control sample. The folate production in yogurt samples showed that Y8 sample has the highest amount of folate (117.93, 38.81 μg/mL determined by microbial and HPLC assay methods, respectively). It was found that the microbial assay method has the ability to measure different forms of folate derivatives compared to the HPLC method. The results exhibited that the isolates examined in this study showed the ability to produce extracellular folate at higher levels and can be used as bio-enrichment cultures to produce functional foods.

Synthesis of di-rhamnolipids by the avirulent, mono-rhamnolipid producing strain Pseudomonas aeruginosa ATCC 9027

To construct a derivative of the avirulent Pseudomonas aeruginosa ATCC 9027 that produces high levels of di-rhamnolipid, that has better physico-chemical characteristics for biotechnological applications than mono-rhamnolipid, which is the sole type produced by ATCC 9027. We used plasmids expressing the rhlC gene, which encodes for rhamnosyl transferase II that transforms mono- to di-rhamnolipids under different promoters and in combination with the gene coding for the RhlR quorum sensing regulator, or the mono-rhamnolipid biosynthetic rhlAB operon. The plasmids tested carrying the rhlC gene under the lac promoter were plasmid prhlC and prhlRC, while prhlAB-R–C expressed this gene from the rhlA promoter, forming part of the artificially constructed rhlAB-R–C operon. We measured rhamnolipds concentrations using the orcinol method and determined the proportion of mono-rhamnolipids and di-rhamnolipids by UPLC/MS/MS. We found that the expression of rhlC in P. aeruginosa ATCC 9027 caused the production of di-rhamnolipids and that the derivative carrying plasmid prhlAB-R–C gives the best results considering total rhamnolipids and a higher proportion of di-rhamnolipids. A P. aeruginosa ATCC 9027 derivative with increased di-rhamnolipids production was developed by expressing plasmid prhlAB-R–C, that produces similar rhamnolipids levels as PAO1 type-strain and presented a higher proportion of di-rhamnolipids than this type-strain.

Metabolic engineering of the substrate utilization pathway in Escherichia coli increases L-lysine production

L-lysine is an essential amino acid with broad applications in the animal feed, human food, and pharmaceutical industries. The fermentation production of L-lysine by Escherichia coli has limitations such as poor substrate utilization efficiency and low saccharide conversion rates. We deleted the global regulatory factor gene mlc and introduced heterologous genes, including the maltose phosphotransferase genes (malAP) from Bacillus subtilis, to enhance the use efficiency of disaccharides and trisaccharides. The engineered strain E. coli XC3 demonstrated improved L-lysine production, yield, and productivity, which reached 160.00 g/L, 63.78%, and 4.44 g/(L‧h), respectively. Furthermore, we overexpressed the glutamate dehydrogenase gene (gdhA) and assimilated nitrate reductase genes (BsnasBC) from B. subtilis, along with nitrite reductase genes (EcnirBD) from E. coli, in strain E. coli XC3. This allowed the construction of E. coli XC4 with a nitrate assimilation pathway. The L-lysine production, yield, and productivity of E. coli XC4 were elevated to 188.00 g/L, 69.44%, and 5.22 g/(L‧h), respectively. After optimization of the residual sugar concentration and carbon to nitrogen ratio, the L-lysine production, yield, and productivity were increased to 204.00 g/L, 72.32%, and 5.67 g/(L‧h), respectively, in a 5 L fermenter. These values represented the increases of 40.69%, 20.03%, and 40.69%, respectively, compared with those of the starting strain XC1. By engineering the substrate utilization pathway, we successfully constructed a high-yield L-lysine producing strain, laying a solid foundation for the industrial production of L-lysine.

Resistance against aminoglycoside antibiotics via drug or target modification enables community-wide antiphage defense.

The ongoing arms race between bacteria and phages has forced bacteria to evolve a sophisticated set of antiphage defense mechanisms that constitute the bacterial immune system. In our previous study, we highlighted the antiphage properties of aminoglycoside antibiotics, which are naturally secreted by Streptomyces. Successful inhibition of phage infection was achieved by addition of pure compounds and supernatants from a natural producer strain emphasizing the potential for community-wide antiphage defense. However, given the dual functionality of these compounds, neighboring bacterial cells require resistance to the antibacterial activity of aminoglycosides to benefit from the protection they confer against phages. In this study, we tested a variety of different aminoglycoside resistance mechanisms acting via drug or target (16S rRNA) modification and demonstrated that they do not interfere with the antiphage properties of the molecules. Furthermore, we confirmed the antiphage impact of aminoglycosides in a community context by coculturing phage-susceptible, apramycin-resistant Streptomyces venezuelae with the apramycin-producing strain Streptoalloteichus tenebrarius. Given the prevalence of aminoglycoside resistance among natural bacterial isolates, this study highlights the ecological relevance of chemical defense via aminoglycosides at the community level.

Genome characterization of Leuconostoc pseudomesenteroides UTNElla29 isolated from Morus nigra (L.) fruits: A promising exopolysaccharides producing strain

Lactic acid bacteria produce various natural polysaccharides that are secreted into the cellular matrix and bonded to the cell surface or released into the surrounding environment. Due to their broad applications in the food industry, the search for novel polysaccharide producer strains remains of interest. In this study, we report the genome sequencing and characterization of Leuconostoc pseudomesenteroides strain UTNElla29 isolated from Morus nigra (L.) fruits. Using in silico different “genome mining” tools we prospect their genetic potential. The genome size was estimated at 2,272,560 bp and a GC content of 39.26%. Of the total proteins, 2170 matched in the EggNOG database. The key genes related to amino acid biosynthesis, carbohydrate metabolism, exopolysaccharide (EPS) production, were annotated. The genome harbors one intact prophage, one CRISPR array, and two putative gene sequences associated with CAS3 Type I, and no plasmids indicating its stability. Moreover, no acquired resistance genes, virulence and pathogenic factors, nor biogenic amines were predicted, suggesting its safety. In vitro preliminary studies showed that EPS yield from 0.4 to 10.2 (g/L) dry biomass and the production varied with the inoculum size and sugar type MRS containing medium. Furthermore, several polyketides and terpenes were predicted, indicating the possibility of generating a wide range of naturally compounds with broad biological functions. However, these promising results suggest that Ella29 is a beneficial strain to be further exploited in vitro for various technological use.

Neuroprotective Effects Exerted by a Combination of Selected Lactic Acid Bacteria in a Mouse Parkinsonism Model under Levodopa-Benserazide Treatment.

Alterations of the microbiota-gut-brain axis has been associated with intestinal and neuronal inflammation in Parkinson's disease (PD). The aim of this work was to study some mechanisms associated with the neuroprotective effect of a combination (MIX) of lactic acid bacteria (LAB) composed by Lactiplantibacillus plantarum CRL2130 (riboflavin overproducing strain), Streptococcus thermophilus CRL808 (folate producer strain), and CRL807 (immunomodulatory strain) in cell cultures and in a chronic model of parkinsonism induced with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in aged mice, and under levodopa-benserazide treatment. In vitro, N2a differentiated neurons were exposed to the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) and treated with intracellular bacterial extracts or with conditioned media from BV-2 cells exposed to the bacterial extracts. In vivo, motor skills, tyrosine hydrolase (TH) in brain and cytokine concentrations in serum and in brain were evaluated. The study of the faecal microbiota and the histology of the small intestine was also performed. The results showed that the neuroprotective effect associated with LAB MIX administration did not interfere with levodopa-benserazide treatment. This effect could be associated with the antioxidant and immunomodulatory potential of the LAB selected in the MIX, and was associated with the significant improvement in the motor tests and a higher number of TH + cells in the brain. In addition, LAB MIX administration was associated with modulation of the immune response. LAB administration decreased intestinal damage with an increase in the villus length /crypt depth ratio. Finally, the administration of the LAB MIX in combination with levodopa-benserazide treatment was able to partially revert the intestinal dysbiosis observed in the model, showing greater similarity to the profiles of healthy controls, and highlighting the increase in the Lactobacillaceae family. Different mechanisms of action would be related to the protective effect of the selected LAB combination which has the potential to be evaluated as an adjuvant for conventional PD therapies.