RESUMO
This study investigated the effect of in situ produced water-soluble α-glucan (LcWSG) and water-insoluble α-glucan (LcWIG) from Leuconostoc citreum SH12 on the physicochemical properties of fermented soymilk. α-Glucans produced by Leuc. citreum SH12 improved water-holding capacity, viscosity, viscoelasticity and texture of fermented soymilk. Gtf1365 and Gtf836 of the five putative glucansucrases were responsible for synthesizing LcWSG and LcWIG during soymilk fermentation, respectively. Co-fermentation of soymilk with Gtf1365 and Gtf836 and non-exopolysaccharide-producing Lactiplantibacillus plantarum D1031 indicated that LcWSG effectively hindered the whey separation of fermented soymilk by increasing viscosity, while LcWIG improved hardness, springiness and accelerated protein coagulation. Fermented soymilk gel formation was mainly based on hydrogen bonding and hydrophobic interactions, which were promoted by both LcWSG and LcWIG. LcWIG has a greater effect on α-helix to ß-sheet translation in fermented soymilk, causing more rapid protein aggregation and thicker cross-linked gel network. Structure-based exploration of LcWSG and LcWIG from Leuc. citreum SH12 revealed their distinct roles in the physicochemical properties of fermented soymilk due to their different ratio of α-1,6 and α-1,3 glucosidic linkages and various side chain length. This study may guide the application of the water-soluble and water-insoluble α-glucans in fermented plant protein foods for their quality improvement.
Assuntos
Fermentação , Glucanos , Leuconostoc , Solubilidade , Leite de Soja , Água , Leuconostoc/metabolismo , Leite de Soja/química , Água/química , Viscosidade , Glucanos/química , Fenômenos QuímicosRESUMO
Daidzein is a major isoflavone compound with an immense pharmaceutical value. This study applied a novel P450 CYP82D26 which can biosynthesize daidzein from (2S)-naringenin. However, the recombinant P450 systems often suffer from low coupling efficiency, leading to an electron transfer efficiency decrease and harmful reactive oxygen species release, thereby compromising their stability and catalytic efficiency. To address these challenges, the SH3-GBD-PDZ (SGP) protein scaffold was applied to assemble a multienzyme system comprising CYP82D26, P450 reductase, and NADP+-dependent aldehyde reductase in desired stoichiometric ratios. Results showed that the coupling efficiency of the P450 system was significantly increased, primarily attributed to the channeling effect of NADPH resulting from the proximity of tethered enzymes and the electrostatic interactions between NADPH and SGP. Assembling this SGP-scaffolded assembly system in Escherichia coli yielded a titer of 240.5 mg/L daidzein with an 86% (2S)-naringenin conversion rate, which showed a 9-fold increase over the free enzymes of the P450 system. These results underscore the potential application of the SGP-scaffolded multienzyme system in enhancing the coupling and catalytic efficiency of the P450 system.
Assuntos
Flavanonas , Isoflavonas , NADPH-Ferri-Hemoproteína Redutase , NADP/metabolismo , NADPH-Ferri-Hemoproteína Redutase/metabolismo , Proteínas , Isoflavonas/metabolismoRESUMO
The active metabolite (S)-equol, derived from daidzein by gut microbiota, exhibits superior antioxidative activity compared with its precursor and plays a vital role in human health. As only 25% to 50% of individuals can naturally produce equol when supplied with isoflavone, we engineered probiotic E. coli Nissle 1917 (EcN) to convert dietary isoflavones into (S)-equol, thus offering a strategy to mimic the gut phenotype of natural (S)-equol producers. However, co-fermentation of EcN-eq with fecal bacteria revealed that gut microbial metabolites decreased NADPH levels, hindering (S)-equol production. Transcriptome analysis showed that the quorum-sensing (QS) transcription factor SdiA negatively regulates NADPH levels and (S)-equol biosynthesis in EcN-eq. Screening AHLs showed that SdiA binding to C10-HSL negatively regulates the pentose phosphate pathway, reducing intracellular NADPH levels in EcN-eq. Molecular docking and dynamics simulations investigated the structural disparities in complexes formed by C10-HSL with SdiA from EcN or E. coli K12. Substituting sdiA_EcN in EcN-eq with sdiA_K12 increased the intracellular NADPH/NADP+ ratio, enhancing (S)-equol production by 47%. These findings elucidate the impact of AHL-QS in the gut microbiota on EcN NADPH metabolism, offering insights for developing (S)-equol-producing EcN probiotics tailored to the gut environment.
RESUMO
Self-assembling peptides are valuable building blocks to fabricate supramolecular biomaterials, which have broad applications from biomedicine to biotechnology. However, limited choices to induce different globular proteins into hydrogels hinder these designs. Here, an easy-to-implement and tunable self-assembling strategy, which employs Ure2 amyloidogenic peptide, are described to induce any target proteins to assemble into supramolecular hydrogels alone or in combination with notable compositional control. Furthermore, the collective effect of nanoscale interactions among amyloid nanofibrils and partially disordered elastomeric polypeptides are investigated. This led to many useful macroscopic material properties simultaneously emerging from one pure protein material, i.e. strong adhesion to any substrates under wet conditions, rapidly self--assembling into robust and porous hydrogels, adaptation to remodeling processes, strongly promoting cell adhesion, proliferation and differentiation. Moreover, he demonstrated this supramolecular material's robust performance in vitro and vivo for tissue engineering, cosmetic and hemostasis applications and exhibited superior performance compared to corresponding commercial counterparts. To the best of his knowledge, few pure protein-based materials could meet such seemingly mutually exclusive properties simultaneously. Such versatility renders this novel supramolecular nanomaterial as next-generation functional protein-based materials, and he demonstrated the sequence level modulation of structural order and disorder as an untapped principle to design new proteins.
Assuntos
Proteínas Amiloidogênicas , Proteínas de Insetos , Nanoestruturas , Peptídeos/química , Nanoestruturas/química , Amiloide/química , Materiais Biocompatíveis/química , Hidrogéis/químicaRESUMO
Medium-chain fatty acids (MCFAs) are essential chemical feedstocks. Microbial production of MCFAs offers an attractive alternative to conventional methods, but the costly media and external inducers limit its practical application. To address this issue and make MCFA production more cost-effective, an E.coli platform was developed using soy whey as a medium and galactose as an autoinducer. We first designed an efficient, stringent, homogeneous, and robust galactose-based autoinduction system for the expression of pathway enzymes by rationally engineering the promoter of the galactose-proton symporter (GalP). Subsequently, the intracellular acetyl-CoA availability and NADH regeneration were enhanced to improve the reversal of the ß-oxidation cycle. The resulting strain yielded 8.20 g/L and 16.42 g/L MCFA in pH-controlled batch fermentation and fed-batch fermentation with glucose added using soy whey as medium, respectively. This study provided a cost-effective and promising platform for MCFA production, as well as future strain development for other value-added chemicals production.
Assuntos
Escherichia coli , Ácidos Graxos , Escherichia coli/metabolismo , Ácidos Graxos/metabolismo , Galactose/metabolismo , Soro do Leite/metabolismo , Análise Custo-Benefício , Engenharia Metabólica/métodos , Proteínas do Soro do Leite/metabolismo , FermentaçãoRESUMO
In this study, Leuconostoc citreum BH10, an endophytic strain, was isolated from aseptically collected xylem sap of birch for the first time, and its exopolysaccharide (LCEPS) production was up to 46.31 g/L in glucan producing medium. The produced LCEPS was purified to obtain two water-soluble fractions, named as LCEPS-1 and LCEPS-2, respectively. The major fraction LCEPS-1 was characterized to be comprised of glucose with average molecular weight of 6.34 × 106 Da. The structure of LCEPS-1 was investigated by spectroscopy analysis, which revealed that LCEPS-1 was identified with containing 90.45 % α-(1,6) linkages in the main chains and 9.55 % α-(1,3) branch linkages. The scanning electron microscope results demonstrated that the dried LCEPS-1 appeared porous surface overlaid with an irregular glittering. The water solubility index (WSI) and water holding capacity (WHC) of LCEPS-1 were 88.02 ± 1.69 % and 241.43 ± 6.38 %, respectively. Besides, it exhibited high thermal stability as well as fine antioxidant activities. Taken together, the results indicated that LCEPS-1 could have good potentiality to be applied in fields of foods, cosmetics, nutraceuticals and pharmaceutical industries as the natural agent.
Assuntos
Betula , Polissacarídeos Bacterianos , Polissacarídeos Bacterianos/química , Glucose , Leuconostoc/química , Água/químicaRESUMO
In this study, the noncovalent interaction mechanisms between soybean 7S globulin and three polyphenols (gallic acid (GA), chlorogenic acid (CA) and (-)-epigallocatechin gallate (EGCG)) were explored and compared using various techniques. Fluorescence experiments showed that GA and EGCG had strong static quenching effects on 7S fluorescence, and that of CA was the result of multiple mechanisms. The interactions caused changes to the secondary and tertiary structure of 7S, and the surface hydrophobicity was decreased. Thermodynamic experiments showed that the combinations of polyphenols with 7S were exothermic processes. Hydrogen bonds and van der Waals forces were the primary driving forces promoting the binding of EGCG and CA to 7S. The combination of GA was mainly affected by electrostatic interaction. The results showed that the structure and molecular weight of polyphenols play an important role in their interactions. This work is helpful for developing products containing polyphenols and soybean protein.
RESUMO
Probiotics have gained significant attention owing to their roles in regulating human health. Recently, spray drying has been considered as a promising technique to produce probiotic powders due to its advantages of high efficiency, cost-saving, and good powder properties. However, the severe environmental conditions from drying and digestion can significantly reduce cell viability, resulting in poor bioaccessibility and bioavailability of live cells. Therefore, there is a need to develop effective targeted delivery systems using spray drying to protect bacteria and to maintain their physiological functions in the targeted sites. This review highlights recent studies about spray-dried targeted delivery vehicles for probiotics, focusing on key strategies to protect bacteria when encountering external stresses, the formation mechanism of particles, the targeted release and colonization mechanisms of live cells in particles with different structures. Advances in the targeted delivery of live probiotics via spray-dried vehicles are still in their early stages. To increase the possibilities for industrialization and commercialization, functional improvement of microcapsules in terms of protection, targeted release, and colonization of bacteria, as well as the effect of spray drying on bacterial physiological functions in the host, need to be further investigated.
RESUMO
The aim of this study is to report the preparation of pectin microspheres by varying degrees of methyl esterification (DM) cross-linked with divalent cationic calcium to encapsulate Lactiplantibacillus plantarum STB1 and L. plantarum LJ1, respectively. Scanning electron microscopy revealed the compact and smooth surface of pectin of DM 28 %, and the stochastic distribution of L. plantarum throughout the gel reticulation. And the pectin of DM 28 % considerably increased probiotics tolerance after continuous exposure to stimulated gastrointestinal tract conditions, with viable counts exceeding 109 CFU/mL. This data indicated that low methoxy-esterification pectin was more efficient to improve the targeted delivery of probiotics in GIT. Additionally, the controlled release of microspheres was dependent on various pH levels. At pH 7.4, the release rates of L. plantarum STB1 and L. plantarum LJ1 reached up to 97.63 % and 95.33 %, respectively. Finally, the Caco-2 cell adhesion model was used to evaluate the cell adhesion rate after encapsulation, which exhibited better adhesion at DM of 60 %.
Assuntos
Lactobacillus plantarum , Probióticos , Humanos , Pectinas/farmacologia , Pectinas/metabolismo , Esterificação , Preparações de Ação Retardada/metabolismo , Microesferas , Células CACO-2 , Colo/metabolismo , Lactobacillus plantarum/metabolismoRESUMO
The prebiotic properties of two purified fractions (GL1-E1 and GL1-E2) of exopolysaccharides (EPSs) from Lacticaseibacillus paracasei GL1 were investigated through in vitro fermentation of pure and human fecal cultures. The results indicated that the simulated digestion under saliva, gastric, and small intestinal conditions had no effect on GL1-E1 and GL1-E2. Additionally, GL1-E1 and GL1-E2 can be used as substrates for Lactobacillus and Lactococcus growth. It was also found that both were gradually degraded and utilized by the gut microbiota. As fermentation proceeded, the pH continued to decrease. Additionally, the total short-chain fatty acid (SCFA) production significantly increased, especially the major SCFA of formic, lactic, and acetic acid. Furthermore, GL1-E1 and GL1-E2 could significantly regulate the composition of the gut microbiota, by increasing the relative abundances of Bacteroides and Phascolarctobacterium, and decreasing the pathogenic bacteria Escherichia-Shigella, Klebsiella, and Fusobacterium. These results suggest that GL1-E1 and GL1-E2 have the potential to be developed as a prebiotic.
Assuntos
Lacticaseibacillus paracasei , Lacticaseibacillus , Humanos , Fermentação , Digestão , Fezes/microbiologia , Ácidos Graxos Voláteis/metabolismo , PrebióticosRESUMO
Oxidative stress is a key process in the development of neurodegenerative diseases. More attention is needed to screen natural antioxidants and explore pharmacological mechanisms. Natural product polysaccharides with no toxic side effects have powerful antioxidant activity. Two purified intracellular polysaccharide fractions (IPS1 and IPS2) from Paecilomyces cicadae TJJ1213 was isolated. Then, a model of H2 O2 -induced oxidative stress in PC12 cells was established to investigate the neuroprotective role of IPS and elucidate the potential protection mechanism. Results showed that IPS1 and IPS2 reduced reactive oxygen species (ROS) production, inhibited the leakage of lactate dehydrogenase (LDH) and Ca2+ and attenuated the expression of apoptotic proteins. In addition, western blots displayed that IPS1 and IPS2 significantly inhibited mitophagy induced by H2 O2 in PC12 cells via PINK/Parkin pathway. Therefore, IPS1 and IPS2 deserved further investigation as protective agents against neurodegenerative diseases.
Assuntos
Fármacos Neuroprotetores , Estresse Oxidativo , Ratos , Animais , Células PC12 , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Polissacarídeos/farmacologia , Apoptose , Fármacos Neuroprotetores/farmacologia , Peróxido de Hidrogênio/farmacologiaRESUMO
(S)-Equol is one of the most bioactive metabolites of the isoflavones with immense nutritional and pharmaceutical value. Soy whey is the major liquid byproduct of the soy product processing industries that is rich in nutrients and (S)-equol biosynthetic precursor daidzin. However, it is usually disposed into the sewage, causing high environmental contamination. Herein, we constructed a recombinant Escherichia coli for the biosynthesis of (S)-equol from soy whey. First, we evaluated daidzin-specific transporters and optimized the anaerobically induced Pnar in the (S)-equol biosynthesis cassette to produce (S)-equol from daidzin. Then, sucrase and α-galactosidase were co-expressed to confer sucrose, stachyose, and raffinose utilization capacity on E. coli. Meanwhile, EIIBCAglc was inactivated to eliminate the daidzin transport inhibition induced by glucose. Finally, combining these strategies and optimizing the fermentation conditions, the optimal strain produced 91.5 mg/L of (S)-equol with a yield of 0.96 mol/mol substrates in concentrated soy whey. Overall, this new strategy is an attractive route to broaden the applications of soy whey and achieve the eco-friendly production of (S)-equol.
RESUMO
3D bioprinting has become a flexible technical means used in many fields. Currently, research on 3D bioprinting is mainly focused on the use of mammalian cells to print organ and tissue models, which has greatly promoted progress in the fields of tissue engineering, regenerative medicine, and pharmaceuticals. In recent years, bacterial bioprinting has gradually become a rapidly developing research fields, with a wide range of potential applications in basic research, biomedicine, bioremediation, and other field. Here, this works reviews new research on bacterial bioprinting, and discuss its future research direction.
Assuntos
Bioimpressão , Animais , Impressão Tridimensional , Engenharia Tecidual , Medicina Regenerativa , Bactérias , Alicerces Teciduais , MamíferosRESUMO
The application of LG-8 and its dry fragments as zearalenone (ZEN) adsorbents was investigated. The study showed that Geotrichum candidum LG-8 and its fragments dried at 55°C or through lyophilization are able to adsorb around 80% of ZEN. However, besides in water and 55°C-drying conditions, SEM indicated that higher 90% of ZEN binding tended to occur when cell walls of fragments were intact with less adhesion among themselves. Notably, ZEN/LG-8 fragments complexes were quite stable, as only 1.262% and 1.969% of ZEN were released after successive pH treatments for 4 h and 5 min. The kinetic data signified that adsorption of ZEN onto LG-8 fragments followed well the pseudo-first-order kinetic model. Isotherm calculations showed Langmuir model was favourable and monolayer adsorption of ZEN occurred at functional binding sites on fragments surface. Therefore, we conclude that it can be an alternative biosorbent to treat water contained with ZEN, since LG-8 is low-cost biomass and its fragments have a considerable high biosorption capacity avoiding impacting final product quality and immunodeficient patients.
RESUMO
Genistin is one of the bioactive isoflavone glucosides found in legumes, which have great nutraceutical and pharmaceutical significance. The market available isoflavones are currently produced by direct plant extraction. However, its low abundance in plant and structural complexity hinders access to this phytopharmaceutical via plant extraction or chemical synthesis. Here, the E. coli cell factory for sustainable production of genistin from glycerol was constructed. First, we rebuilt the precursor genistein biosynthesis pathway in E. coli, and its titer was then increased by 668% by identifying rate-limiting steps and applying an artificial protein scaffold system. Then de novo production of genistin from glycerol was achieved by functional screening and introduction of glycosyl-transferases, UDP-glucose pathway and specific genistin efflux pumps, and 48.1 mg/L of genistin was obtained. A further engineered E. coli strain equipped with an improved malonyl-CoA pathway, alternative glycerol-utilization pathways, acetyl-CoA carboxylase (ACC), and CRISPR interference (CRISPRi) mediated regulation produced up to 137.8 mg/L of genistin in shake flask cultures. Finally, 202.7 mg/L genistin was achieved through fed-batch fermentation in a 3-L bioreactor. This study represents the de novo genistin production from glycerol for the first time and will lay the foundation for low-cost microbial production of glucoside isoflavones. In addition, the multiphase workflow may provide a reference for engineering the biosynthetic pathways in other microbial hosts as well, for green manufacturing of complex natural products.
Assuntos
Escherichia coli , Isoflavonas , Escherichia coli/genética , Escherichia coli/metabolismo , Engenharia Metabólica , Glicerol/metabolismo , Isoflavonas/metabolismo , GlucosídeosRESUMO
Two exopolysaccharide fractions (GL1-E1 and GL1-E2) of Lacticaseibacillus paracasei GL1 were isolated with the molecular weights of 3.9 × 105 Da and 8.2 × 105 Da, respectively. Both fractions possessed mannose, glucose, and galactose in molar ratios of 1.16:1.00:0.1, and 3.81:1.00:0.12, respectively. A structural arrangement of two fractions was proposed by methylation, one-dimensional and two-dimensional nuclear magnetic resonance experiments. The backbone of GL1-E1 consisted of â4)-α-D-Glcp(1â, â3,4)-α-D-Manp(1â, â3,6)-α-D-Manp(1â, â6)-α-D-Manp(1â, and â6)-α-D-Galp(1â with α-D-Glcp at branching point. The backbone of GL1-E2 consisted of â4)-α-D-Glcp(1â, â3,4)-α-D-Manp(1â, â3,6)-α-D-Manp(1â, â6)-α-D-Manp(1â, â6)-α-D-Galp(1â, and â4)-ß-D-Manp(1â, and the side chain also consisted of α-D-Manp residue. In addition, the differential scanning calorimetry (DSC) analysis indicated that both GL1-E1 and GL1-E2 had good thermal stability. Furthermore, the two fractions could promote the viability of RAW264.7 cells and exert an immunomodulatory role by enhancing phagocytosis, increasing nitric oxide (NO) release and promoting the expression of cytokines.
RESUMO
Transcriptomic analysis was used to investigate the antibacterial mechanism of phenolic compounds from kefir fermented soy whey (FSP) against Escherichia coli 0157:H7 and Listeria monocytogenes. The kefir fermentation increased the concentration of several phenolic aglycones with proven antibacterial efficacy in the FSP. The time-kill curve showed that 2× MICs of the FSP killed >99.9 % of the strains within 2 h of exposure. The checkerboard fractional inhibition concentration (FIC) assay proved that phenolics were the sole antibacterial agent in the FSP. The transmission electron microscope (TEM) photomicrograph corroborated the propidium iodide (PI) uptake, protein, and nucleic acid leakage assays. They demonstrated that the phenolics permeated the cell membrane, disrupted the cytoplasm, and caused cell lysis in the treated cells leading to protein and nucleic acid leakage. The transcriptome analysis revealed that exposure of the cells to MICs of the phenolics induced molecular responses leading to differential expression of 1850 genes in E. coli 0157:H7 and 2090 in L. monocytogenes. The phenolics suppressed the expression of genes crucial for carbohydrate utilization, transmembrane glucose transport, tricarboxylic acid (TCA), and ATP synthesis. The phenolic-induced stress also downregulated the expression of quorum sensing and virulence-related genes, peptidoglycan and phospholipid synthases, and ABC transporters. The cells initiated a resistance response by stimulating the two-component signal transduction systems to trigger the over-expression of a cascade of genes involved in stress resistance, gluconeogenesis, ATPase activity and proton transmembrane transport. Nonetheless, the data indicated that the phenolics suppressed the expression of translational proteins that would have facilitated the resistance and repair of the cell damage caused by the phenolics. The study provides discrete data evidence that FSP could be used to control the pathogenicity and the proliferation of E. coli 0157:H7 and L. monocytogenes in our foods and food systems.
Assuntos
Escherichia coli O157 , Kefir , Listeria monocytogenes , Ácidos Nucleicos , Listeria monocytogenes/fisiologia , Escherichia coli O157/fisiologia , Soro do Leite , Microbiologia de Alimentos , Propídio , Peptidoglicano , Prótons , Transcriptoma , Antibacterianos/farmacologia , Trifosfato de Adenosina , Perfilação da Expressão Gênica , Adenosina Trifosfatases , Transportadores de Cassetes de Ligação de ATP , Fosfolipídeos , Glucose , Ácidos Tricarboxílicos , Contagem de Colônia MicrobianaRESUMO
Exopolysaccharide (EPS) isolated from Lactobacillus helveticus SNA12 were purified, and one fraction (SNA12-EPS) was obtained. The structure of SNA12-EPS was proposed using Fourier-transform infrared spectroscopy, gas chromatography-mass spectrometry, and nuclear magnetic resonance. Results showed that SNA12-EPS was rich in galactose and glucose with the molar ratios of 2.1:1.0, and SNA12-EPS possessed the repeat units of â3)-ß-D-Glcp-(1â3)-ß-D-Glcp-(1â4)-ß-D-Galp-(1â with an average molecular weight of 3.81 × 105 Da. The scanning electron microscope results showed that SNA12-EPS had a tight structure with a smooth and uneven surface. Furthermore, the prebiotic potential of SNA12-EPS was performed using in vitro simulated digestion with human faecal fermentation. SNA12-EPS was not digested by digestive juice, and it could markedly regulate the gut microbiota composition by increasing the relative abundances of Parabacteroides and Blautia and decreasing the abundance of pathogenic bacteria of Fusobacterium. Additionally, SNA12-EPS improved the ability of gut microbiota to produce short-chain fatty acids.
Assuntos
Lactobacillus helveticus , Carboidratos da Dieta , Fermentação , Humanos , Peso Molecular , Polissacarídeos Bacterianos/químicaRESUMO
The effects of high exopolysaccharides (EPS) - producing Lactobacillus helveticus MB2-1 on the structure and storage stability of set yoghurts, and the interactions between its EPS (molecular weight 9.34 × 104 Da) from Sayram ketteki yoghurt (SKY) and sodium caseinate (CAS) were studied. The rheology, microstructure, texture and storage stability of the three set yoghurts including control yoghurt (Control), adding-probiotic yoghurt (APY) and SKY were investigated, which showed that the SKY exhibited less shear thinning than the Control and APY, and the textural indexes and storage stability of the SKY were significantly better than that of other two yoghurts (p < 0.05). Moreover, the increased turbidity, decreased ζ potential and surface hydrophobicity of EPS/CAS complex coacervation were determined at EPS/CAS mass ratio of 3 (corresponding to 0.33 g/L of CAS and 1 g/L of EPS), mainly owing to the electrostatic attraction of the two biopolymers to form aggregates. Besides, the higher sizes and more aggregation of EPS/CAS complexes were formed at pH 3.5. Taken together, the results indicated that the high EPS-producing characteristic of L. helveticus MB2-1 could positively influence the qualities of set yoghurts, and the EPS/CAS complex coacervation in dairy products was closely related to the EPS/CAS mass ratio and pH condition.
Assuntos
Lactobacillus helveticus , Caseínas , Fermentação , Lactobacillus helveticus/química , Peso Molecular , Reologia , IogurteRESUMO
In this work, we reported an in situ exopolysaccharide (in situ-EPS1) containing rare fucose produced by Lactobacillus helveticus MB2-1 in Sayram ketteki yoghurt, which made it unique. Its fine structure was characterized by GPC, HPLC, FT-IR, GC-MS,1HNMR and 13CNMR together with two-dimensional (2D) NMR spectra. The results revealed that in situ-EPS1 was a new heteropolysaccharide with molecular weight of 1.06 × 105 Da, and was composed of mannose, rhamnose, glucose, galactose and fucose with the following repeating units. Furthermore, the in situ-EPS1 exhibited significant antibiofilm effect against Methicillin-resistant Staphylococcus aureus (MRSA). Notably, the in situ-EPS1 did not interfere with the planktonic growth of MRSA strain, whereas inhibited its cell metabolic activity and the transcription of genes related to biofilm formation. This unique antibiofilm but non-antibacterial mechanism supposedly prevented the development of bacterial drug resistance, which may open a new door to fight against these drug-resistant microorganisms.