scholarly journals Purification and characterization of novel exopolysaccharides produced from Lactobacillus paraplantarum KM1 isolated from human milk and its cytotoxicity

Author(s):  
Kanika Sharma ◽  
Nivedita Sharma ◽  
Shweta Handa ◽  
Shruti Pathania

Abstract Background Microbial origin polysaccharides have gained popularity due to lesser toxicity, better degradability and selectivity as compared to their synthetic counterparts and can be used as emulsifier, stabilizer, thickener, texturizer, flocculating and gelling agent. Here main emphasis on exopolysaccharide production from potential lactic acid bacteria that has GRAS status. Results This work was aimed at isolating, purifying and characterizing an extracellular polysaccharide (EPS) produced by a foodgrade lactic acid bacteria Lactobacillus paraplantarum KM1. L. paraplantarum KM1 was isolated from human milk and identified by conventional and molecular techniques. The 16S rRNA sequence of the isolate was registered in National Centre for Biotechnology Information (NCBI) under accession number KX671558. L. paraplantarum KM1 was found to produce EPSs in lactose containing MRS medium, and the maximum yield (47.4 mg/ml) was achieved after 32-h incubation. As evident from TLC and HPLC analyses, the polysaccharide was found to be a heteropolymer-containing glucose, galactose and mannose as main sugars. Different oligosaccharides namely hexoses were obtained after partial hydrolysis of the polymer using MALDI-ToF-MS. The total molecular weight of all polysaccharides present was 348.7 kDa with 100 °C thermal stability as well as water soluble in nature. Cell cytotoxicity revealed that the purified EPS was safe for consumption; thus, it can be used in various food industries as emulsifying and texture agent. Conclusions The present study highlighted that exopolysaccharides could be harnessed to improve food products in terms of texture, emulsifying agents, pharmaceutical industry (antioxidants, antitumour, anti-inflammatory and antiviral agents) and as safety purposes.

2020 ◽  
Vol 84 (1) ◽  
pp. 63-72
Author(s):  
QING ZHANG ◽  
XIAOJUAN SONG ◽  
WENLIN SUN ◽  
CHAN WANG ◽  
CUIQIN LI ◽  
...  

ABSTRACT A total of 115 isolates of lactic acid bacteria were screened from traditional fermented foods in Guizhou Province, People's Republic of China. The cholesterol removal rates of 86 isolates ranged from 7.29 to 25.66%, and 18 isolates showed a cholesterol removal rate of more than 15%. According to the results of physiological and biological tests, 13 isolates were selected to determine the fermentation performance; 9 isolates—MT-4, MT-2, PJ-15, SR2-2, SQ-4, SQ-7, ST2-2, ST2-6, and NR1-7—had high tolerance of bile salt and acid and had a survival rate of more than 96% under pH 3.0 and 0.3% bile salt. ST2-2, SR2-2, NR1-7, SQ-4, and MT-4 had high survival rate in different concentrations of NaCl and NaNO2 under different temperatures. According to BLAST comparison results of the 16S rRNA sequence in the GenBank database and the genetic distance of the 16S rRNA sequence with an ortho-connected algorithm, SR2-2, NR1-7, and ST2-2 were identified as Lactobacillus plantarum, MT-4 was identified as Lactobacillus pentosus, and SQ-4 was identified as Lactobacillus paraplantarum. Moreover, strains SQ-4 and MT-4 were added to fermented beef. Results showed that the fermented beef had delicious taste and was popular to consumers because of its proper pH, pleasant colors, high viable cell count, and suitable content of bound and immobilized water. These results provide a basis for the development of new starter formulation for the production of high-quality fermented meat products. HIGHLIGHTS


2016 ◽  
Vol 62 (6) ◽  
pp. 514-524 ◽  
Author(s):  
Sandra Rayén Quilodrán-Vega ◽  
Julio Villena ◽  
José Valdebenito ◽  
María José Salas ◽  
Cristian Parra ◽  
...  

Probiotics are usually isolated from the gastrointestinal tract of humans and animals. The search of probiotics in human milk is a recent field of research, as the existence of the human milk microbiome was discovered only about a decade ago. To our knowledge, no reports regarding the potential probiotic effect of bacteria from swine milk have been published. In this work, we isolated several lactic acid bacteria from swine milk and evaluated them for them potential as probiotics. Among the isolated strains, Lactobacillus curvatus TUCO-5E showed antagonistic effects against swine-associated gastrointestinal pathogens. TUCO-5E was able to reduce the growth of enterotoxigenic and enterohemorrhagic Escherichia coli strains as well as pathogenic salmonella. In vitro exclusion and displacement assays in intestinal epithelial cells showed a remarkable antagonistic effect for L. curvatus TUCO-5E against Salmonella sp. strain TUCO-I7 and Salmonella enterica ATCC 13096. Moreover, by using a mouse model of Salmonella infection, we were able to demonstrate that preventative administration of L. curvatus TUCO-5E for 5 consecutive days was capable of decreasing the number of Salmonella enterica serovar Typhimurium in the liver and spleen of treated mice, compared with the controls, and prevented dissemination of the pathogen to the blood stream. Therefore, we have demonstrated here that swine milk is an interesting source of beneficial bacteria. In addition, the results of this work suggest that L. curvatus TUCO-5E is a good candidate to study in vivo the protective effect of probiotics against intestinal infection and damage induced by Salmonella infection in the porcine host.


2021 ◽  
Vol 12 ◽  
Author(s):  
Sihan You ◽  
Shuai Du ◽  
Gentu Ge ◽  
Tao Wan ◽  
Yushan Jia

This study aimed to isolate and identify lactic acid bacteria (LAB) from the native grass and naturally fermented silage from the Mongolian Plateau. The effect of selected strains on bacterial community and quality of native grass silage was also studied. Strains XM2, 265, and 842 could grow normally at 15°C–30°C, pH 4.0–8.0, and NaCl 3 and 6.5%; they were identified as Lactiplantibacillus plantarum subsp. plantarum, Pediococcus acidilactici, and Latilactobacillus graminis, by sequencing 16S rRNA, respectively. The three strains (XM2, 265, and 842) and one commercial additive (L) were used as inoculants and singularly added to the native grass. Compared to the control, the dry matter content was significantly (p < 0.05) lower in L and XM2 groups. The water-soluble carbohydrate content was significantly (p < 0.05) higher in control than in other groups. Compared with the control, the crude protein and ammonia nitrogen contents were significantly (p < 0.05) higher and lower in the LAB-treated groups, and the acid and detergent fiber contents were significantly (p < 0.05) reduced in the L and XM2 groups than those in other groups. There was a significant (p < 0.05) difference in the pH value, lactic acid content, and lactic acid-to-acetic acid ratio in L and XM2 groups than in other groups. Compared with the control, the number of LAB was significantly (p < 0.05) higher in LAB-treated silages, whereas no significant (p > 0.05) differences were observed in yeast and aerobic bacteria in all groups. Compared to the control, the Shannon index was significantly (p < 0.05) reduced. Simpson and Chao1 were significantly (p < 0.05) increased. Principal coordinate analysis based on the unweighted UniFrac distance showed clear separation of the bacterial community in fresh materials and LAB-treated silages. Besides, compared to the control, the principal coordinate analysis of LAB-treated silages was also separate. After 30 days of fermentation, the relative abundance of Firmicutes increased and was the primary phylum in all silages. Compared with the control, the abundance of Firmicutes and Proteobacteriawas significantly (p < 0.05) higher and lower in L and XM2 groups. In contrast, no significant differences were observed among control, 265, and 842 groups. At the genus level, the relative abundance of Lactobacillus, Enterobacter, Pediococcus, and Weissella was increased and dominated the native grass fermentation. Compared with the control, the abundance of Lactobacillus was significantly (p < 0.05) higher in L, XM2, and 842 groups, while no significant (p > 0.05) differences were observed between the control and 265 groups. The abundance of Pediococcus was higher than that in other groups. Consequently, the results demonstrated that LAB significantly influenced silage fermentation by reconstructing microbiota, and Lactobacillus was the dominant genus in the native grass silages. Furthermore, the results showed that strain XM2 could effectively improve the silage quality, and it is considered a potential starter for the native grass silage.


Molecules ◽  
2020 ◽  
Vol 25 (7) ◽  
pp. 1560 ◽  
Author(s):  
Huaidan Zhang ◽  
Jeremy N. Marchant-Forde ◽  
Xinyi Zhang ◽  
Yan Wang

NH3 emission has become one of the key factors for aerobic composting of animal manure. It has been reported that adding microbial agents during aerobic composting can reduce NH3 emissions. However, environmental factors have a considerable influence on the activity and stability of the microbial agent. Therefore, this study used cornstalk biochar as carriers to find out the better biological immobilization method to examine the mitigation ability and mechanism of NH3 production from laying hen manure composting. The results from different immobilized methods showed that NH3 was reduced by 12.43%, 5.53%, 14.57%, and 22.61% in the cornstalk biochar group, free load bacteria group, mixed load bacteria group, and separate load bacteria group, respectively. Under the simulated composting condition, NH3 production was 46.52, 38.14, 39.08, and 30.81 g in the treatment of the control, mixed bacteria, cornstalk biochar, and cornstalk biochar separate load immobilized mixed bacteria, respectively. The cornstalk biochar separate load immobilized mixed bacteria treatment significantly reduced NH3 emission compared with the other treatments (p < 0.05). Compared with the control, adding cornstalk biochar immobilized mixed bacteria significantly decreased the electrical conductivity, water-soluble carbon, total nitrogen loss, and concentration of ammonium nitrogen (p < 0.05), and significantly increased the seed germination rate, total number of microorganisms, and relative abundance of lactic acid bacteria throughout the composting process (p < 0.05). Therefore, the reason for the low NH3 emission might be due not only to the adsorption of the cornstalk biochar but also because of the role of complex bacteria, which increases the relative abundance of lactic acid bacteria and promotes the acid production of lactic acid bacteria to reduce NH3 emissions. This result revealed the potential of using biological immobilization technology to reduce NH3 emissions during laying hen manure composting.


Agronomy ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1432
Author(s):  
Horst Auerbach ◽  
Peter Theobald

Whole-crop rye harvested before maturity represents a valuable forage for silage production. Due to the scarcity of data on fermentation characteristics and aerobic stability (ASTA) and the lack of information on mycotoxin formation during aeration of early-cut rye (ECR) silage after silo opening, we evaluated the effects of different additive types and compositions. Wilted forage was treated with various biological and chemical additives, ensiled in 1.5-L glass jars and stored for 64 days. Fermentation pattern, yeast and mould counts and ASTA were determined at silo opening. In total 34 mycotoxins were analysed in wilted forage and in silage before and after 240 h of air exposure. Chemical additives caused the lowest dry matter (DM) losses during fermentation accompanied with the lowest ethanol production and the highest water-soluble carbohydrate concentration. Aerobic deterioration, which started within two days after silo opening in silage left untreated and inoculated with homofermentative lactic acid bacteria, was prevented by the combined use of hetero- and homofermentative lactic acid bacteria and the chemical additive containing sodium nitrite, hexamethylene tetramine and potassium sorbate. Moreover, these two additives largely restricted the formation of the mycotoxin roquefortine C to < 0.05 mg kg−1 DM after aeration, whereas untreated silage contained 85.2 mg kg−1 DM.


2000 ◽  
Vol 66 (5) ◽  
pp. 2224-2226 ◽  
Author(s):  
Frédéric Ampe

ABSTRACT Based on 16S rRNA sequence comparison, we have designed a 20-mer oligonucleotide that targets a region specific to the speciesLactobacillus manihotivorans recently isolated from sour cassava fermentation. The probe recognized the rRNA obtained from all the L. manihotivorans strains tested but did not recognize 56 strains of microorganisms from culture collections or directly isolated from sour cassava, including 29 species of lactic acid bacteria. This probe was then successfully used in quantitative RNA blots and demonstrated the importance of L. manihotivoransin the fermentation of sour cassava starch, which could represent up to 20% of total lactic acid bacteria.


2016 ◽  
Vol 142 ◽  
pp. 91-97 ◽  
Author(s):  
Marianne Øksnes Dalheim ◽  
Nina Bjørk Arnfinnsdottir ◽  
Göran Widmalm ◽  
Bjørn E. Christensen

2003 ◽  
Vol 66 (3) ◽  
pp. 355-363 ◽  
Author(s):  
M. M. BRASHEARS ◽  
D. JARONI ◽  
J. TRIMBLE

Lactic acid bacteria (LAB) were selected on the basis of characteristics indicating that they would be good candidates for a competitive exclusion product (CEP) that would inhibit Escherichia coli O157:H7 in the intestinal tract of live cattle. Fecal samples from cattle that were culture negative for E. coli O157:H7 were collected. LAB were isolated from cattle feces by repeated plating on deMan Rogosa Sharpe agar and lactobacillus selection agar. Six hundred eighty-six pure colonies were isolated, and an agar spot test was used to test each isolate for its inhibition of a four-strain mixture of E. coli O157:H7. Three hundred fifty-five isolates (52%) showed significant inhibition. Seventy-five isolates showing maximum inhibition were screened for acid and bile tolerance. Most isolates were tolerant of acid at pH levels of 2, 4, 5, and 7 and at bile levels of 0.05, 0.15, and 0.3% (oxgall) and were subsequently identified with the API system. Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus delbreukii, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus cellobiosus, Leuconostoc spp., and Pediococcus acidilactici were the most commonly identified LAB. Nineteen strains were further tested for antibiotic resistance and inhibition of E. coli O157:H7 in manure and rumen fluid. Four of these 19 strains showed susceptibility to all of the antibiotics, 13 significantly reduced E. coli counts in manure, and 15 significantly reduced E. coli counts in rumen fluid (P &lt; 0.05) during at least one of the sampling periods. One of the strains, M35, was selected as the best candidate for a CEP. A 16S rRNA sequence analysis of M35 revealed its close homology to Lactobacillus crispatus. The CEP developed will be used in cattle-feeding trials.


2020 ◽  
Author(s):  
Blanca Rosa Aguilar Uscanga ◽  
Ariana Rodríguez Arreola ◽  
Josué R. Solís Pacheco ◽  
Monique Lacroix ◽  
Edgar Balcazar López ◽  
...  

1938 ◽  
Vol 16b (2) ◽  
pp. 46-53 ◽  
Author(s):  
Blythe Alfred Eagles ◽  
Olga Okulitch ◽  
Arthur Stephen Kadzielawa

The influence of three distinct activators prepared from tomatoes, yeast, or liver, on the metabolism of two species of lactic acid bacteria has been studied. One of these activators is Bios II A, and the other two have been shown to be components constituting Bios II B. On the basis of their physical and chemical properties, it is suggested that the growth stimulants required by the lactic acid bacteria are identical with certain of the heat-stable accessory food factors of the Vitamin-B complex essential for the growth of animals.


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