RESUMO
BACKGROUND: The excessive production and accumulation of melanin in the epidermal skin layer can result in skin hyperpigmentation and darkening. Current technologies for regulating melanin are based on inhibiting melanin biosynthesis. They have low effectiveness and safety issues. AIMS: This study aimed to evaluate the potential role of Pediococcus acidilactici PMC48 as a probiotic strain in medicines and cosmetics for skin treatment. MATERIALS AND METHODS: Meanwhile, our research team has reported that P. acidilactici PMC48 strain isolated from sesame leaf kimchi can directly decompose the already synthesized melanin. It can also inhibit melanin biosynthesis. In the present study, we investigated the skin-whitening effect of this strain by arranging an 8-week clinical trial with 22 participants. PMC48 was applied to each participant's artificially UV-induced tanned skin in the clinical trial. Its whitening effect was investigated based on visual evaluation, skin brightness, and melanin index. RESULTS: PMC48 showed a significant effect on the artificially induced pigmented skin. The color intensity of the tanned skin was decreased by 47.647%, and skin brightness was increased by 8.098% after the treatment period. PMC48 also significantly decreased the melanin index by 11.818%, indicating its tyrosinase inhibition capacity. Also, PMC48 improved skin moisture content level by 20.943%. Additionally, 16S rRNA-based amplicon sequencing analysis showed a distinct increase in Lactobacillaceae in the skin by up to 11.2% at the family level without affecting other skin microbiota. Furthermore, it showed no toxicity in in vitro or in vivo analyses. DISCUSSION: These results indicate that P. acidilactici PMC48 is a promising probiotic strain that can be used to develop medicines and cosmetic products to solve skin-related problems. CONCLUSIONS: These results demonstrate that P. acidilactici PMC48 can be a potential probiotic for the cosmetic industry against different skin disorders.
Assuntos
Cosméticos , Hiperpigmentação , Pediococcus acidilactici , Humanos , Pediococcus acidilactici/genética , Melaninas , RNA Ribossômico 16S , Pele , Hiperpigmentação/tratamento farmacológico , Cosméticos/farmacologiaRESUMO
Gamma-aminobutyric acid (GABA) is a crucial inhibitory neurotransmitter in the sympathetic nervous system that exerts regulatory effects on the blood, immune, and nervous systems. GABA production in som-fak, a traditional fermented fish of Thailand, has been attributed to the activity of lactic acid bacteria (LAB). The present study aims to characterize the LAB isolates and compare the genomes and GABA synthesis genes of selected isolates capable of GABA production. Thirteen isolates demonstrating GABA synthesis capability were identified based on their phenotypic and genotypic characteristics. Seven isolates (group I: LSF3-3, LSF8-3, LSF9-1, LSF9-3, LSF9-6, LSF9-7, and LSF10-14) were identified as Levilactobacillus brevis with 99.78-100% similarity. LSF2-1, LSF3-2, LSF5-4, and LSF6-5 (group II) were identified as Lactiplantibacillus pentosus with 99.86-100% similarity. Strain LSF1-1 (group III) was identified as Pediococcus acidilactici (99.47%), and LSF10-4 (group IV) was identified as Pediococcus pentosaceus with 99.93% similarity. The GABA production of isolates ranged from 0.087 to 16.935 g/L. The maximum production of 16.935 g/L from 3% monosodium glutamate was obtained from strain LSF9-1. Gene and genome analysis revealed that L. brevis LSF9-1 has multiple gad genes in the genome, such as gadB1, gadB2, gadC1, and gadC2, making it the potential strain for GABA production. Additionally, the genome analysis of P. acidilactici LSF1-1 consists of gadA, gadB, and gadC, which respond to controlling GABA production and export. Furthermore, strain LSF1-1 was considered safe, containing no virulence factors. Thus, Levilactobacillus brevis LSF9-1 and Pediococcus acidilactici LSF1-1 have the potential for GABA production and probiotic use in future studies.
Assuntos
Levilactobacillus brevis , Pediococcus acidilactici , Pediococcus acidilactici/genética , Ácido gama-AminobutíricoRESUMO
The GRAS (generally recognized as safe) strain Pediococcus acidilactici is well known for its antibacterial and probiotic functions. Furthermore, as P. acidilactici has excellent high temperature and salt resistance, it is an ideal host for the production of food enzymes, food additives, and pharmaceuticals. In this regard, it is desirable and feasible to enhance the production of these products through the metabolic engineering of P. acidilactici. However, the rare gene expression elements greatly obstruct the development of engineering P. acidilactici. In this study, we screened and constructed a library of promoter-5'-UTR (PUTR) complexes in P. acidilactici DY15 for regulating gene expression at the transcription and translation levels. In the post-log phase, the mRNA and protein expression level ranges of the 90 screened native PUTRs were 0.059-2010% and 0.77-245%, respectively, of the P32 promoter. Besides, several PUTRs exhibited great expression stability under high temperature, salt, and ethanol stress. We analyzed the structure of PUTRs and obtained the conserved regions of the promoter and 5'-UTR. Based on the identified core regions of PUTRs, we constructed a panel of combinatorial PUTRs with higher and stable protein expression levels. The strongest combinatorial PUTR was 853% of the P32 promoter in the protein expression level. Finally, the obtained PUTRs were applied to optimize the expression level of aminotransferase and improve the phenyllactic acid (PLA) production in P. acidilactici DY15. The achieved yield was 950.6 mg/L, which was 79.2% higher than the wild-type strain. These results indicated that the obtained PUTRs with gradient strength had great potential for precisely regulating gene expression to achieve various goals in P. acidilactici.
Assuntos
Pediococcus acidilactici , Probióticos , Pediococcus acidilactici/genética , Pediococcus acidilactici/metabolismo , Regiões Promotoras Genéticas/genética , Engenharia Metabólica , Antibacterianos/metabolismoRESUMO
Lactic acid bacteria are the well acknowledged probiotics that can cure a variety of diseases. In this study, we observed the in vivo potentials of Pediococcus to treat hyperglycemia, hypercholesterolemia and gastrointestinal infections. A total of 77 Lactobacillus were isolated from the milk of 10 cows and 10 goats, four of those strains inhibited both carbohydrates-hydrolyzing enzymes, α-glucosidase, and α-amylase. They all showed antagonistic effects on pathogenic E. coli and S. Typhimurium which were confirmed by performing pathogen challenge test and visualizing on Electron microscopy. 16S rRNA gene sequence identified that all four strains belong to Pediococcus genus which were further distinguished as Pediococcus acidilactici by pheS gene sequence. Whole genome sequence analysis revealed their non-pathogenic properties for human and the presence of probiotic genes responsible for stress resistance, immunomodulation, adhesion, metal and drug resistance. In vivo trial with diabetes-induced mice ascertained that all Pediococcus acidilactici had significant potentials to reduce elevated glucose and low-density lipoprotein level in blood. Interestingly, two out of four strains were significantly more effective (p < 0.0001 each) than metformin in reducing the blood glucose level. This in vivo study demonstrated that Pediococcus acidilactici might be a promising probiotic to prevent hyperglycemia, hypercholesterolemia and gastrointestinal infections.
Assuntos
Doenças Transmissíveis , Gastroenteropatias , Hipercolesterolemia , Hiperglicemia , Pediococcus acidilactici , Probióticos , Feminino , Humanos , Bovinos , Camundongos , Animais , Pediococcus acidilactici/genética , RNA Ribossômico 16S/genética , Escherichia coli , Genômica , Hiperglicemia/prevenção & controle , Probióticos/farmacologia , Probióticos/uso terapêutico , Pediococcus/genética , CabrasRESUMO
Lactic acid bacteria are indispensable functional microorganisms for cereal vinegar brewing, but cell activities are inhibited by the dominant acetic acid stress. Herein, an acetic-acid-tolerant strain isolated previously was identified as Pediococcus acidilactici, which also exhibited good resistance to other stresses during vinegar brewing. Proteomics analysis evidenced that differentially expressed proteins involved in the glycolysis and gluconeogenesis pathway, pyruvate metabolism, and sugar phosphotransferase system were all downregulated. Meanwhile, saturation of fatty acids and antioxidant enzymes was strengthened. The effects of several proteins on the resistance of P. acidilactici and Lactobacillus lactis relied on the types of strain and stress. AccA and AcpP participating in fatty acid metabolism and biosynthesis and Mnc related to stress response were found to protect cells by modifying fatty acid compositions and reinforcing the antioxidant defense system. Our works deepen the mechanisms of P. acidilactici under acetic acid and offer targets for engineering cell tolerance.
Assuntos
Pediococcus acidilactici , Ácido Acético/metabolismo , Antioxidantes/metabolismo , Fermentação , Pediococcus/genética , Pediococcus/metabolismo , Pediococcus acidilactici/genética , Pediococcus acidilactici/metabolismo , Fosfotransferases/metabolismo , Proteômica , Piruvatos/metabolismo , Açúcares/metabolismoRESUMO
Pediococcus acidilactici is commonly used for pediocin production and lactic acid fermentation. However, a high-efficiency genome editing tool is unavailable for this species. In this study, we constructed endogenous subtype II-A CRISPR-Cas system-based genome interference plasmids which carried a "repeat-spacer-repeat" cassette in the pMG36e shuttle vector. These plasmids exhibited self-interference activities in P. acidilactici LA412. Then, the genome-editing plasmids were constructed by cloning the upstream/downstream donor DNA into the corresponding interference plasmids to exert high-efficiency markerless gene deletion, gene integration, and point mutation in P. acidilactici LA412. We found that endogenous CRISPR-mediated depletion of the native plasmids enhanced the cell growth and that integration of an l-lactate dehydrogenase gene into the chromosome enhanced both cell growth and lactic acid production. IMPORTANCE A rapid and precise genome editing tool will promote the practical application of Pediococcus acidilactici, one type of lactic acid bacterium with excellent stress tolerance and probiotic characteristics. This study established a high-efficiency endogenous CRISPR-Cas system-based genome editing tool for P. acidilactici and achieved different genetic manipulations, including gene deletion, gene insertion, mononucleotide mutation, and endogenous plasmid depletion. The engineered strain edited by this tool showed significant advantages in cell growth and lactic acid fermentation. Therefore, our tool can satisfy the requirements for genetic manipulations of P. acidilactici, thus making it a sophisticated chassis species for synthetic biology and bioindustry.
Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Ácido Láctico/metabolismo , Pediococcus acidilactici , Proteínas de Bactérias/genética , Fermentação , L-Lactato Desidrogenase/genética , Engenharia Metabólica , Pediococcus acidilactici/genética , Pediococcus acidilactici/crescimento & desenvolvimento , Pediococcus acidilactici/metabolismo , Mutação PuntualRESUMO
Our aim was to isolate, identify and characterize probiotic bacteria as vitamin producers in particular B2 and B9. 150 human fecal samples were collected and used for isolation of vitamin producers-probiotics. 49 isolates were chosen for screening their genome by PCR for the presence of riboflavin and folic acid genes. As a result, three isolates were selected and their production of the B2 and B9 were confirmed by HPLC. The three isolates were identified on species level by sequencing their 16S rRNA gene which showed 100% identical to strains of Pediococcus acidilactici. Thus, they were named as P. acidilactici WNYM01, P. acidilactici WNYM02, P. acidilactici WNYM03 and submitted to the Genbank database with accession numbers. They met the probiotic criteria by expressing 90-95% survival rate at pH (2.0-9.0) and bile salt up to 2% for 3 h in addition to their antimicrobial activity against gram positive and negative microorganisms. They also showed no hemolytic activity and common pattern for antibiotic susceptibility. Our three strains were tested individually or in mixture in vivo on rat colitis model compared to ulcerative group. The strains were administrated orally to rats in daily dose containing CFU 109 for 14 days then followed by induction of colitis using acetic acid then the oral administration was continued for more four days. The histology results, the anti-inflammatory and anti-oxidative stress biomarkers showed the protective role of the strains compared to the ulcerative group. As a conclusion, we introduce novel three probiotic candidates for pharmaceutical preparations and health applications.
Assuntos
Colite/terapia , Ácido Fólico/metabolismo , Microbioma Gastrointestinal , Pediococcus acidilactici/fisiologia , Probióticos/administração & dosagem , Riboflavina/metabolismo , Ácido Acético/toxicidade , Administração Oral , Animais , Antibacterianos/farmacologia , Ácidos e Sais Biliares/farmacologia , Colite/induzido quimicamente , Colo/patologia , Modelos Animais de Doenças , Fezes/microbiologia , Ácido Fólico/análise , Humanos , Concentração de Íons de Hidrogênio , Masculino , Pediococcus acidilactici/efeitos dos fármacos , Pediococcus acidilactici/genética , Pediococcus acidilactici/isolamento & purificação , RNA Ribossômico 16S/análise , RNA Ribossômico 16S/metabolismo , Ratos , Ratos Wistar , Riboflavina/análiseRESUMO
Pediococcus acidilactici is a reliable bacteriocin producer and a promising probiotic species with wide application in the food and health industry. However, the underlying genetic features of this species have not been analyzed. In this study, we performed a comprehensive comparative genomic analysis of 41 P. acidilactici strains from various ecological niches. The bacteriocin production of 41 strains were predicted and three kinds of bacteriocin encoding genes were identified in 11 P. acidilactici strains, namely pediocin PA-1, enterolysin A, and colicin-B. Moreover, whole-genome analysis showed a high genetic diversity within the population, mainly related to a large proportion of variable genomes, mobile elements, and hypothetical genes obtained through horizontal gene transfer. In addition, comparative genomics also facilitated the genetic explanation of the adaptation for host environment, which specify the protection mechanism against the invasion of foreign DNA (i.e. CRISPR/Cas locus), as well as carbohydrate fermentation. The 41 strains of P. acidilactici can metabolize a variety of carbon sources, which enhances the adaptability of this species and survival in different environments. This study evaluated the antibacterial ability, genome evolution, and ecological flexibility of P. acidilactici from the perspective of genetics and provides strong supporting evidence for its industrial development and application.
Assuntos
Genoma Bacteriano , Pediococcus acidilactici/genética , Antibacterianos/biossíntese , Bacteriocinas/biossíntese , Genômica , Pediococcus acidilactici/classificação , Pediococcus acidilactici/metabolismoRESUMO
Fermentative production of sodium lactate generally is a low efficient process because of the high Na+ osmatic stress on lactic acid bacterium cells. In this study, the homogeneous genes encoding Na+/H+ antiporters were screened and overexpressed in Pediococcus acidilactici for the enhancement of Na+ transmembrane transportation. The function of the gene RS02775 was identified and its overexpressing in P. acidilactici resulted in the significantly improved sodium lactate production. The recombinant not only accelerated the sugar consumption, but also achieved the record high titer of sodium lactate by 121.1 g/L using pure sugars and 132.4 g/L using wheat straw. The transcription analysis shows that the overexpression of Na+/H+ antiporter significantly upregulated the transcription of the sugar phosphorylation genes of P. acidilactici under high Na+ stress. This study provides an effective method for high titer production of sodium lactate using both pure sugars and lignocellulose feedstocks.
Assuntos
Pediococcus acidilactici , Metabolismo dos Carboidratos , Fermentação , Ácido Láctico , Pediococcus/genética , Pediococcus acidilactici/genética , Lactato de SódioRESUMO
p-Benzoquinone (BQ) is a lignin-derived inhibitor to microbial strains. Unlike the furan inhibitors, p-benzoquinone is recalcitrant to traditional detoxification methods. This study shows a biological degradation of p-benzoquinone and a simultaneous D-lactic acid fermentation by an engineered Pediococcus acidilactici strain. The overexpression of an oxidoreductase gene ZMO1116 from Zymomonas mobilis encoding oxidoreductase was identified to improve the D-lactic acid fermentability of P. acidilactici against p-benzoquinone. The gene ZMO1116 was integrated into the genome of P. acidilactici and enabled the engineered P. acidilactici to convert p-benzoquinone into less toxic hydroquinone (HQ), resulting in the improved p-benzoquinone tolerance. Simultaneous saccharification and co-fermentation (SSCF) was conducted using the pretreated and biodetoxified corn stover containing p-benzoquinone, the D-lactic acid production of the engineered strain (123.8 g/L) was 21.4 % higher than the parental strain (102.0 g/L). This study provides a practical method on robust p-benzoquinone tolerance and efficient cellulosic chiral lactic acid fermentation from lignocellulose feedstock.
Assuntos
Benzoquinonas/metabolismo , Fermentação , Ácido Láctico/metabolismo , Oxirredutases/genética , Pediococcus acidilactici/genética , Biodegradação Ambiental , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Lignina/metabolismo , Oxirredutases/metabolismo , Pediococcus acidilactici/metabolismo , Zea mays/metabolismo , Zea mays/microbiologia , Zymomonas/genética , Zymomonas/metabolismoRESUMO
Overproduction and accumulation of melanin in the skin will darken the skin and cause skin disorders. So far, components that can inhibit tyrosinase, a melanin synthase of melanocytes, have been developed and used as ingredients of cosmetics or pharmaceutical products. However, most of existing substances can only inhibit the biosynthesis of melanin while melanin that is already synthesized and deposited is not directly decomposed. Thus, their effects in decreasing melanin concentration in the skin are weak. To overcome the limitation of existing therapeutic agents, we started to develop a substance that could directly biodegrade melanin. We screened traditional fermented food microorganisms for their abilities to direct biodegrade melanin. As a result, we found that a kimchi-derived Pediococcus acidilactici PMC48 had a direct melanin-degrading effect. This PMC48 strain is a new strain, different from P. acidilactici strains reported so far. It not only directly degrades melanin, but also has tyrosinase-inhibiting effect. It has a direct melanindecomposition effect. It exceeds existing melanin synthesis-inhibiting technology. It is expected to be of high value as a raw material for melanin degradation drugs and cosmetics.
Assuntos
Alimentos Fermentados/microbiologia , Melaninas/biossíntese , Pediococcus acidilactici/isolamento & purificação , Pediococcus acidilactici/metabolismo , Perilla/microbiologia , Folhas de Planta/microbiologia , Sobrevivência Celular/efeitos dos fármacos , Melanócitos/efeitos dos fármacos , Monofenol Mono-Oxigenase , Pediococcus acidilactici/genética , Filogenia , RNA Ribossômico 16S , República da CoreiaRESUMO
The aim of this research was to isolate bacteria capable of biotransforming daidzein from fresh feces from pregnant horses. A Hungate anaerobic roller tube was used for anaerobic culture. Single colonies were picked at random and incubated with daidzein. High performance liquid chromatography was used to detect whether the isolated bacteria were able to biotransform the substrate. A strain capable of reducing daidzein was selected and characterized using sequence analysis of 16S rDNA, and a phylogenetic tree was constructed. The morphological physiological and biochemical characteristics of the strain were investigated. A facultative anaerobic, Gram-positive bacterium capable of converting daidzein to dihydrodaidzein was isolated and named HXBM408 (MF992210). A BLAST search of HXBM408's 16S rDNA sequence against the GenBank database suggested that the strain has 99% similarity with Pediococcus acidilactici strain DSM (NR042057). The morphological, physiological, and biochemical characteristics of HXBM408 are very similar to those of Pediococcus. Based on these characteristics, the strain was identified as Pediococcus acidilactici. The bacterial strain HXBM408 isolated from the feces of pregnant horses was able to reduce the isoflavone daidzein to dihydrodaidzein.
Assuntos
Bactérias/isolamento & purificação , Bactérias/metabolismo , Cavalos/metabolismo , Cavalos/microbiologia , Isoflavonas/metabolismo , Pediococcus acidilactici/isolamento & purificação , Pediococcus acidilactici/metabolismo , Animais , Bactérias/genética , Biotransformação , DNA Bacteriano/genética , DNA Ribossômico/genética , Fezes/microbiologia , Feminino , Pediococcus acidilactici/genética , Filogenia , Fitoestrógenos/metabolismo , Gravidez , RNA Ribossômico 16S/genética , Glycine max/químicaRESUMO
Pediococcus acidilactici NCDC 252 is a facultative anaerobe of dairy origin that possessed all studied in vitro probiotic attributes and several useful enzyme activities. Its whole genome was sequenced and analysed for its evolutionary relationship with other lactic acid bacteria (LAB). This is a novel sequence and first report of genome sequence of P. acidilactici of dairy origin. Its genome is relatively larger than other studied genomes of P. acidilactici and is comprised of 40 scaffolds that totals to 3,243,337 bases and 44.5% GC content. A total of 3054 coding sequences (CDS) were identified by RAST and DIAMOND servers. The genome also encoded different enzyme activities required for utilization of various carbohydrates. This was also confirmed by carbohydrate utilization studies. The genome also encoded genes for probiotics properties. The phylogenetic analysis of P. acidilactici NCDC 252 genome was done using Maximum Parsimony and Maximum Likelihood methods to study its evolution and relatedness to other LABs based upon their 16S rDNA sequences. The strain exhibited highest resemblance to Lactobacillus plantarum WCFS1 and is also much close to P. acidilactici based on similarity of ribosomal protein. The strain seems to have acquired some genes for its adaptation in dairy/environmental niche. This genome sequence is novel with genome more similar to L. plantarum and biochemical and phenotypic characteristics of P. acidilactici.
Assuntos
Pediococcus acidilactici/enzimologia , Pediococcus acidilactici/genética , Pediococcus acidilactici/metabolismo , Evolução Biológica , DNA Ribossômico , Evolução Molecular , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Ácido Láctico/metabolismo , Lactobacillales/genética , Lactobacillales/metabolismo , Redes e Vias Metabólicas , Pediococcus/genética , Filogenia , ProbióticosRESUMO
Engineering D-lactic acid dehydrogenases for higher activity on various 2-oxo acids is important for the synthesis of 2-hydroxy acids that can be utilized in a wide range of industrial fields including the production of biopolymers, pharmaceuticals, and cosmetic compounds. Although there are many D-lactate dehydrogenases (D-LDH) available from a diverse range of sources, there is a lack of biocatalysts with high activities for 2-oxo acids with large functional group at C3. In this study, the D-LDH from Pediococcus acidilactici was rationally designed and further engineered by controlling the intermolecular interactions between substrates and the surrounding residues via analysis of the active site structure of D-LDH. As a result, Y51L mutant with the catalytic efficiency on phenylpyruvate of 2200 s-1 mM-1 and Y51F mutant on 2-oxobutryate and 3-methyl-2-oxobutyrate of 37.2 and 23.2 s-1 mM-1 were found, which were 138-, 8.5-, and 26-fold increases than the wild type on the substrates, respectively. Structural analysis revealed that the distance and the nature of the interactions between the side chain of residue 51 and the substrate C3 substituent group significantly affected the kinetic parameters. Bioconversion of phenyllactate as a practical example of production of the 2-hydroxy acids was investigated, and the Y51F mutant presented the highest productivity in in vitro conversion of D-PLA.
Assuntos
Substituição de Aminoácidos , Proteínas de Bactérias/química , Biocatálise , Butiratos/química , Hemiterpenos/química , Cetoácidos/química , L-Lactato Desidrogenase/química , Pediococcus acidilactici/enzimologia , Proteínas de Bactérias/genética , L-Lactato Desidrogenase/genética , Mutação de Sentido Incorreto , Pediococcus acidilactici/genéticaRESUMO
Xylan oligosaccharides (XOS) are the hydrolysates of xylan. To compare the proliferation effect of XOS, glucose, fructo oligosaccharides (FOS), xylose, XOS, and a media without carbohydrate source (control) on Pediococcus acidilactici strain BCC-1, the de novo sequencing of Pediococcus acidilactici strain BCC-1 was conducted, and the underlying mechanism of prebiotic xylo oligosaccharide between xylose and XOS was revealed through transcriptomic analysis. Compared to FOS, glucose, and xylose, XOS exhibits a good performance in promoting the fermentation of Pediococcus acidilactici BCC-1. The genome of Pediococcus acidilactici BCC-1 revealed genes encoding XOS transportation and utilized related enzymes, including ATP-binding cassette (ABC) transporters, arabinofuranosidase, xylanase, xylosidase, xylose isomerase, and xylulose kinase. Transcriptome analysis showed that XOS treatment enhanced genes involving carbohydrate metabolism, an ABC transporter sugar system, pentose and glucuronate interconversions, pyruvate metabolism, and the TCA process when compared to xylose treatment. It suggested XOS treatment enhanced sugar absorption and utilization. These results are useful in the understanding of the metabolic pathway of XOS in Pediococcus acidilactici BBC-1 and may contribute to the optimization of the probiotic effect of Pediococcus acidilactici BCC-1 as novel complex feed additives.
Assuntos
Proteínas de Bactérias/genética , Oligossacarídeos/metabolismo , Pediococcus acidilactici/genética , Xilanos/metabolismo , Proteínas de Bactérias/metabolismo , Perfilação da Expressão Gênica , Pediococcus acidilactici/metabolismo , Prebióticos/análiseRESUMO
The aim of the present study was isolation and molecular identification of lactic acid bacteria from King grass and their application to improve the fermentation quality of sweet Sorghum. Seventy-six strains of LAB were isolated; five strains were selected for Physiological and morphological tests and 16S rRNA sequencing. All five strains grew at different pH 3.5-8.0, different temperature 35, 40, 45, 50 °C and different NaCl concentrations 3, 6.5, 9.5%. Strains HDASK were identified Lactobacillus plantarum and SK3907, SK2A32, SK3A42 and ASKDD Pediococcus acidilactici. Three isolated strains and one commercial strain were added to sweet sorghum. Silage was prepared of four treatments and one control with three replicates as control (SKC, adding 2 ml/kg sterilizing water), L. plantarum commercial bacteria (SKP), L. plantarum (HDASK) isolated from King grass (SKA), P. acidilactici (SK3907) isolated from King grass (SKB) and P. acidilactici (ASKDD) isolated from King grass (SKD). All silage were prepared using polyethylene terephthalate bottles, and incubated at room temperature for different ensiling days. The level of pH, acetic acid, NH3-N, water soluble carbohydrate and butyric acid was significantly (P < 0.05) decreased. Lactic acid, ethanol and propionic acid (PA) was significantly (P < 0.05) increased in treatments compared to control. The dry matter, propionic acid neutral detergent fiber, acid detergent fiber did not significantly (P < 0.05) differ among the treatments but the values were increased and decreased. The number of yeast, mold and LAB were significantly (P < 0.05). It is suggested that the supplementation of LAB could enhanced the fermentation quality of sweet Sorghum silage.
Assuntos
Fermentação , Lactobacillales/genética , Lactobacillales/isolamento & purificação , Poaceae/microbiologia , Sorghum , Ácido Acético/análise , Ácido Butírico/análise , Metabolismo dos Carboidratos , DNA Bacteriano , Etanol/análise , Fungos/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio , Ácido Láctico/análise , Ácido Láctico/metabolismo , Lactobacillales/classificação , Lactobacillales/fisiologia , Lactobacillus plantarum/genética , Lactobacillus plantarum/crescimento & desenvolvimento , Lactobacillus plantarum/isolamento & purificação , Lactobacillus plantarum/fisiologia , Pediococcus acidilactici/genética , Pediococcus acidilactici/crescimento & desenvolvimento , Pediococcus acidilactici/isolamento & purificação , Pediococcus acidilactici/fisiologia , Filogenia , Propionatos/análise , RNA Ribossômico 16S/genética , Silagem/microbiologia , Cloreto de Sódio/metabolismo , TemperaturaRESUMO
Abstract Pediococcus acidilactici strain S1, a lactic acid-fermenting bacterium, was isolated from makgeolli-a Korean traditional fermented alcoholic beverage. Here we report the 1,980,172 bp (G + C content, 42%) genome sequence of Pediococcus acidilactici strain S1 with 1,525 protein-coding sequences (CDS), of which 47% could be assigned to recognized functional genes. The genome sequence of the strain S1 might provide insights into the genetic basis of the lactic acid bacterium with alcohol-tolerant.
Assuntos
Genoma Bacteriano , Ácido Láctico/metabolismo , Bebidas Alcoólicas/microbiologia , Pediococcus acidilactici/isolamento & purificação , Pediococcus acidilactici/genética , Sequência de Bases , República da Coreia , Fermentação , Pediococcus acidilactici/metabolismo , Sequenciamento Completo do GenomaRESUMO
Pediococcus acidilactici is a widely used probiotic, and Salmonella enterica serovar Gallinarum (SG) is a significant pathogen in the poultry industry. In this study, we improved the antimicrobial activity of P. acidilactici against SG using UV mutation and genome shuffling (GS). To improve antimicrobial activity against SG, UV mutagenesis was performed against wild-type P. acidilactici (WT), and five mutants showed improved antimicrobial activity. To further improve antimicrobial activity, GS was performed on five UV mutants. Following GS, four mutants showed improved antimicrobial activity compared with the UV mutants and WT. The antimicrobial activity of GS1 was highest among the mutants; however, the activity was reduced when the culture supernatant was treated with proteinase K, suggesting that the improved antimicrobial activity is due to a proteinous substance such as bacteriocin. To validate the activity of GS1 in vivo, we designed multi-species probiotics and performed broiler feeding experiments. Groups consisted of no treatment (NC), avilamycin-treated (PC), probiotic group 1 containing WT (T1), and probiotic group 2 containing GS1 (T2). In broiler feeding experiments, coliform bacteria were significantly reduced in T2 compared with NC, PC, and T1. The cecal microbiota was modulated and pathogenic bacteria were reduced by GS1 oral administration. In this study, GS1 showed improved antimicrobial activity against SG in vitro and reduced pathogenic bacteria in a broiler feeding experiment. These results suggest that GS1 can serve as an efficient probiotic, as an alternative to antibiotics in the poultry industry.
Assuntos
Antibiose , Embaralhamento de DNA , Mutagênese , Pediococcus acidilactici/genética , Pediococcus acidilactici/fisiologia , Probióticos , Salmonella/fisiologia , Animais , Antibacterianos/farmacologia , Anti-Infecciosos , Bacteriocinas/biossíntese , Bacteriocinas/farmacologia , Ceco/microbiologia , Galinhas/microbiologia , Meios de Cultura/química , Endopeptidase K/metabolismo , Genoma Bacteriano , Pediococcus acidilactici/efeitos dos fármacos , Pediococcus acidilactici/efeitos da radiação , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/terapia , Probióticos/química , Salmonella/efeitos dos fármacos , Salmonelose Animal/microbiologia , Salmonelose Animal/terapiaRESUMO
Biotechnological production of vanillin is gaining momentum as the natural synthesis of vanillin that is very expensive. Ferulic acid (FA), a costly compound, is used as the substrate to produce vanillin biotechnologically and the making process is still expensive. Therefore, this study investigated the practical use of an agrobiomass waste, rice bran, and provides the first evidence of a cost-effective production of vanillin within 24 h of incubation using recombinant Pediococcus acidilactici BD16 (fcs +/ech +). Introduction of two genes encoding feruloyl CoA synthetase and enoyl CoA hydratase into the native strain increased vanillin yield to 4.01 g L-1. Bioconversion was monitored through the transformation of phenolic compounds. A hypothetical metabolic pathway of rice bran during the vanillin bioconversion was proposed with the inserted pathway from ferulic acid to vanillin and compared with that of other metabolic engineered strains. These results could be a gateway of using recombinant lactic acid bacteria for industrial production of vanillin from agricultural waste.
Assuntos
Agricultura , Benzaldeídos/metabolismo , Biomassa , Biotransformação , Oryza , Pediococcus acidilactici/genética , Biotecnologia/economia , Biotecnologia/métodos , Enoil-CoA Hidratase/genética , Microbiologia Industrial/economia , Microbiologia Industrial/métodos , Engenharia Metabólica/economia , Engenharia Metabólica/métodos , Redes e Vias Metabólicas , Oryza/química , Pediococcus acidilactici/metabolismo , Fenóis/química , Fenóis/metabolismoRESUMO
Pediococcus acidilactici strain S1, a lactic acid-fermenting bacterium, was isolated from makgeolli-a Korean traditional fermented alcoholic beverage. Here we report the 1,980,172bp (G+C content, 42%) genome sequence of Pediococcus acidilactici strain S1 with 1,525 protein-coding sequences (CDS), of which 47% could be assigned to recognized functional genes. The genome sequence of the strain S1 might provide insights into the genetic basis of the lactic acid bacterium with alcohol-tolerant.