Lactic Acid Bacterium Enterococcus Faecium Research Articles

Saccharina latissima, candy-factory waste, and digestate from full-scale biogas plant as alternative carbohydrate and nutrient sources for lactic acid production

To substitute petroleum-based materials with bio-based alternatives, microbial fermentation combined with inexpensive biomass is suggested. In this study Saccharina latissima hydrolysate, candy-factory waste, and digestate from full-scale biogas plant were explored as substrates for lactic acid production. The lactic acid bacteria Enterococcus faecium, Lactobacillus plantarum, and Pediococcus pentosaceus were tested as starter cultures. Sugars released from seaweed hydrolysate and candy-waste were successfully utilized by the studied bacterial strains. Additionally, seaweed hydrolysate and digestate served as nutrient supplements supporting microbial fermentation. According to the highest achieved relative lactic acid production, a scaled-up co-fermentation of candy-waste and digestate was performed. Lactic acid reached a concentration of 65.65 g/L, with 61.69% relative lactic acid production, and 1.37 g/L/hour productivity. The findings indicate that lactic acid can be successfully produced from low-cost industrial residues.

Efecto de la fecha de corte y del uso de aditivos en la composición química y calidad fermentativa de ensilado de girasol

El objetivo fue evaluar fechas de corte y del uso de aditivos sobre la calidad del ensilado de la planta entera de girasol. La variedad forrajera (Rumbosol-91) se cosechó en las semanas 1, 3 y 5 post-floración (F1, F2 y F3, respectivamente) y tratada con los siguientes aditivos: 1) 1.5 × 105 ufc de inoculante g-1 de forraje, a base de bacterias lácticas homofermentativas Enterococcus faecium, Pediococcus pentosaceus y Lactobacillus plantarum (INOC), 2) 3 ml kg-1 de forraje de una solución al 85% de ácido fórmico (FORM) y 3) sin aditivo (Testigo); siguiendo un diseño factorial 3x3 con cinco repeticiones. La producción de efluente y las pérdidas totales de materia seca (MS) se redujeron, desde 282 y 134 g kg-1 en F+1 hasta 96 y 87 g kg-1 en F+5 como resultado del alto contenido de humedad del forraje próxima a la floración. El análisis NIRS de las muestras de ensilaje mostró que los contenidos de proteína, fibra y digestibilidad descendían significativamente con la madurez de la planta; la rápida acumulación de aceite en la MS hizo que la concentración energética fuese superior en el estado fenológico más avanzado. La calidad fermentativa de los ensilajes fue satisfactoria, independientemente del momento de corte y del uso de aditivo. Se concluye que es preferible el corte de la planta a las cinco semanas post-floración, donde se espera una fermentación aceptable sin necesidad de conservantes.

Utilizing Gelatinized Starchy Waste from Rice Noodle Factory as Substrate for L(+)-Lactic Acid Production by Amylolytic Lactic Acid Bacterium Enterococcus faecium K-1.

To valorize starchy waste from rice noodle factory, bioconversion of gelatinized starchy waste (GSW) to value-added product as L(+)-lactic acid, the monomer for polylactate synthesis, was investigated using amylolytic lactic acid bacterium, Enterococcus faecium K-1. Screening for appropriate nitrogen source to replace expensive organic nitrogen sources revealed that corn steep liquor (CSL) was the most suitable regarding high efficacy for L(+)-LA achievement and low-cost property. The successful applying statistic experimental design, Plackett-Burman design incorporated with central composite design (CCD), predicted the maximum L(+)-LA of 93.07g/L from the optimized medium (OM) containing 125.7g/L GSW and 207.3g/L CSL supplemented with CH3COONa, MgSO4, MnSO4, K2HPO4, CaCl2, (NH4)2HC6H5O7, and Tween80. Minimizing the medium cost by removal of all inorganic salts and Tween80 from OM was not an effect on L(+)-LA yield. Fermentation using the optimized medium without minerals (OM-Mi) containing only GSW (125.7g/L) and CSL (207.3g/L) in a 10-L fermenter was also successful. Thinning GSW with α-amylase from Lactobacillus plantarum S21 increased L(+)-LA productivity in the early stage of 24-h fermentation. Not only showing the feasible bioconversion process for GSW utilizing as a substrate for L(+)-LA production, this research also demonstrated the efficient model for industrial starchy waste valorization.

INVESTIGATION OF VARIOUS WAYS TO STABILIZE LACTIC ACID BACTERIA CELLS DURING LONG-TERM STORAGE

Various methods of stabilizing the cells of the lactic acid bacterium Enterococcus faecium during long-term storage were studied. It is shown that the long-term survival of E. faecium is promoted by the addition of Enterosgel, Polysorb, the use of LB substratum, as well as their immobilization in humic-silanol gel.

Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum

Gene encoding cyclomaltodextrinase (Cdx) from amylolytic lactic acid bacterium Enterococcus faecium K-1 was cloned and nucleotide sequence was analyzed. The open-reading frame consisted of 1767bp encoding 588 deduced amino acids. Consequently, four typically conserved regions of the glycoside hydrolase family 13 were revealed; however, nine exceeding amino acids (DSYQMTDVP) were found at the 282–290 position in comparison to previously reported cyclomaltodextrinases. This difference is believed to have an influence on the substrate specificity of this enzyme. The recombinant CDases expressed in Escherichia coli BL21 (CDX_E) and Lactobacillus plantarum WCFS1 (CDX_L) with high expression levels of 8041 and 5511U/L were purified by Ni-NTA affinity chromatography. The active form CDX is a dimeric protein with two identical subunits of 62kDa, approximately. Both CDX_E and CDX_L revealed nearly similar properties, but the thermostability of CDX_L was slightly higher. Mn2+ and Co2+ at a concentration of 1mM stimulated the enzyme activity, while the Ag+, Cu2+ and SDS solution completely inhibited enzyme activity. CDX exhibited the highest activity with α-cyclodextrin and β-cyclodextrin, but lower toward pullulan and starch. Importantly, this is the first report describing genes, the molecular structure and properties of cyclomaltodextrinase derived from lactic acid bacteria E. faecium.

Effect of cryopreservation and microencapsulation of lactic acid bacterium Enterococcus faecium MC13 for long-term storage

The aim of this work was to investigate the effect of cryoprotectants on the survival of probiotic bacterium Enterococcus faecium MC13 during freeze drying and storage. The maximum relative cell viabilities were observed when cells were freeze dried and stored at −20 °C, which is optimum temperature for the preservation of E. faecium. At all storage temperatures, trehalose was found to be retaining the highest relative cell viability than other cryoprotectants. In addition, alginate–chitosan capsules were produced to encapsulate E. faecium with the aim of enhancing survival of probiotic cells and keeping the probiotic during exposure to the harsh gastro-intestinal conditions. Encapsulation of probiotic into alginate–chitosan capsules found to be retaining higher survival of probiotic cells (4.342 ± 0.26 Log CFU mL −1) at −20 °C for six months. Microencapsulated cells were resistant to simulated gastric (pH 2.0) and intestinal fluids (pH 7.5), resulting in significantly enhanced survival when compared with free cells. During in vivo treatment, capsules were broken and probiotic cells were directly released into the intestinal tract of rat. This result showed that microencapsulation of E. faecium MC13 with alginate and a chitosan coating offers an effective means of delivery of viable cells to the colon and maintains their survival during the adverse gastro-intestinal conditions.

Optimization of Conditions for the Maximum Bacteriocin Production of Enterococcus faecium DB1 Using Response Surface Methodology

The bacteriocin-producing lactic acid bacteria Enterococcus faecium DB1 was isolated from Korean traditional gajami sikhae. Culture conditions were optimized by response surface methodology (RSM) to maximize bacteriocin DB1 production. E. faecium DB1 displayed the highest bacteriocin activity when grown in modified MRS medium containing sucrose, rather than glucose, as a carbon source. The effects of temperature, initial pH, and sucrose concentration were tested to determine the optimum conditions for maximum bacteriocin production by E. faecium DB1. A central composite design was used to control the three variables in the experiment. RSM revealed that the optimum values for bacteriocin production were 27.66 g/L sucrose, temperature of <TEX>$34.37^{\circ}C$</TEX>, and an initial pH of 6.54. A 2.08-fold increase in bacteriocin production was obtained with sucrose-containing MRS medium compared to production in standard MRS medium.

Enhancement of egg yolk color by paprika combined with a probiotic

Enhancing egg yolk color by feeding laying hens paprika extract (Color-Up, Kohkin Chemical Co. Ltd., Higashiosaka, Japan; 5 g of xanthophylls/kg) and potentially accelerating the process by concurrent feeding of a probiotic (Balantol powder, Kohkin Chemical Co. Ltd.; 107 of lactic acid bacterium Enterococcus faecium BIO-4R/g) was examined. A total of 144 twenty-five-week-old laying hens were randomly assigned to 3 groups: a basal diet group (17.5% CP, 2,750 kcal of ME/kg; control), the basal diet plus 0.1% dietary paprika group, and the basal diet plus 0.1% dietary paprika plus 0.1% probiotic group. Each group consisted of 4 replicates with 12 birds. Hen egg production was recorded daily from 25 to 34 wk of age. Twelve eggs per diet group were evaluated weekly for egg quality. No differences were found in any parameter measured except for egg yolk color. Egg yolk color scores were greater in the paprika group (11.69) than in the control group (9.71; P < 0.05). The egg yolk color scores of the paprika plus probiotic group (12.00) were not significantly different from those of the paprika group. We concluded that paprika extract improved egg yolk color, whereas the probiotic combined with paprika extract did not affect egg yolk color at a level of 107 cfu/g.