Search results for: Lactococcus lactis

Authors: Mehdi Ariana, Javad Hamedi

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Nisin is a commercial bacteriocin that is used as a food preservative and produced by Lactococcus lactis subsp. lactis. Nisin production through co-culture fermentation can be performed for increasing nisin quantities. Since lactate accumulation in the fermentation medium can prevent L. lactis growth and therefore reduce nisin production, the simultaneous culture of microorganisms can enhance L. lactis growth by a reduction in the amount of lactic acid. In this study, conducted coculture of L.lactis subsp. lactic and the yeast Yarrowia lipolytica. Both strains are cultured in a molasses-based medium that is mainly constructed of sucrose. Y. lipolytica is not able to use sucrose as a carbon source but is able to consume lactate and decrease lactic acid in the medium. So, Lactic acid consumption can increase pH value and stimulate L. lactis growth. The results showed the mixed culture increased L. lactis growth 6 times higher than that of pure culture and could enhance nisin activity by up to 40%.

Keywords: co-culture fermentation, lactococcus lactis subsp lactis, yarrowia lipolytica, nisin

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Authors: Maha Alaoui Ismaili, Bouchta Saidi, Mohamed Zahar, Abed Hamama

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112 lactic isolates were obtained from 15 samples of camel raw milk produced in Laayoune Boujdour Sakia-El Hamra region (South of Morocco). The main objective was the identification of species of lactic flora belonging to Lactococcus, Lactobacillus and Leuconostoc. Data obtained showed predominance of cocci among lactic isolates (86.6%) while lactic rods represented only 13.4%. With regard to genera identified, Enterococcus was the mostly found out (53.57%), followed by Lactococcus (28.57%), Lactobacillus (13.4%) and Leuconostoc (4.4 %). Identification of the lactic isolates according to their morphological, physiological, and biochemical characteristics led to differentiating 11 species with Lactococcus lactis ssp lactis biovar diacetylactis being the mostly encountered (24.1%) followed by Lactobacillus brevis (3.57%), Lactobacillus plantarum (3.57%), Lactobacillus delbrueckii subsp lactis (3.57%) and Lactococcus lactis subsp cremoris (2.67%).

Keywords: raw camel milk, south of morocco, lactic acid bacteria, identification

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Authors: Souid Wafa, Boudjenah-Haroun Saliha, Khacef Linda

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In this work, we focused on the valuation of discharges from the dairy industry. Whey is by-product of dairy industry, which is a formidable pollution factor and contains components (lactose, minerals and proteins) with high nutritional value. Whey is an excellent culture medium for microorganisms. The objective of our work is to investigate the ability of a lactic strain (of the genus Lactococcus) to grow in culture media based on whey of cattle and camels and comparing it with that recorded on M17 as indicator medium. In this study we isolated from a local sample of camel milk a lactic strain (S1).the strain had positive Gram shaped, cocci form and catalase (-). The strain has been purified by the method of streaks on M17 medium. Phenotypic identification allows us to classify this strain in the species: Lactococcus lactis subsp. Cremoris. We subsequently tested the ability of this strain to grow in cattle whey medium and camel whey, both media were deproteinized and unsupplemented. The obtained results revealed that: The cattle and camel whey are appropriate media for the growth of the strain Lactococcus lactis subsp cremoris but is more adapted to grow on a medium rich in lactose as the camel whey. In fact, after 48h and at initial pH 6.8 this strain acidified more camel whey (pH 3.99) than cattle whey (pH 4.8). And biomass produced in the camel whey is 1.50g /1 by contributing to the cattle whey which is 1g / l.

Keywords: cremoris, dairy industry, Lactococcus lactis subsp, medium, whey

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Authors: M. H. Nahaisi, N. M. Almaroum

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Laban is a Libyan traditional fermented milk product. This lactic fermentation has been known in many cities of Libya long time ago as stable, nutritious, refreshing drink especially during the summer. 16 naturally fermented milk samples were collected from different cities located in North West of Libya. The average pH, titratable acidity, fat and total solids were 4.16, 0.73%, 1.54% and 8.12 % respectively. Coliform, yeast and mold counts were 21×10⁴, 39×10⁴ and 41 ×10³ cfu/ ml. respectively. The average Lactococcus, Streptococcus, Mesophilic Lactobacillus / Leuconostoc and Thermophilic Lactobacillus counts were 99 ×10⁷, 96 ×10⁷, 93 ×10⁷ and 15 ×10⁷ cfu / ml. respectively. A total of one hundred forty two lactic acid bacteria (LAB) isolates were identified to the genus level as Lactobacillus (48.59%), Lactococcus (43.66%), Streptococcus (4.93%) and Leuconostoc (2.82%). Sugar fermentation tests have revealed that the most frequently Lactobacillus species was found to be Lactobacillus delbrueckii ssp. lactis (62.32%) followed by Lactobacillus plantarum (31.88%). Furthermore, other selected LAB isolates were identified by API 50 CH test as Lactococcus lactis ssp. lactics, Lactobacillus pentosus, Lactobacillus brevis and Leuconostoc mesenteroides ssp. cremoris.

Keywords: traditional fermented milk, laban, lactococcus, streptococcus, mesophilic lactobacillus, thermophilic lactobacillus counts

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Authors: Ankita Singh, Chandan Gorain, Amirul I. Mallick

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Successful adherence of the mucosal epithelial cells is the key early step for Campylobacter jejuni pathogenesis (C. jejuni). A set of Surface Exposed Colonization Proteins (SECPs) are among the major factors involved in host cell adherence and invasion of C. jejuni. Among them, constitutively expressed surface-exposed lipoprotein adhesin of C. jejuni, JlpA, interacts with intestinal heat shock protein 90 (hsp90α) and contributes in disease progression by triggering pro-inflammatory response via activation of NF-κB and p38 MAP kinase pathway. Together with its ability to express in the bacterial surface, higher sequence conservation and predicted predominance of several B cells epitopes, JlpA protein reserves its potential to become an effective vaccine candidate against wide range of Campylobacter sps including C. jejuni. Given that chickens are the primary sources for C. jejuni and persistent gut colonization remain as major cause for foodborne pathogenesis to humans, present study explicitly used chickens as model to test the immune-protective efficacy of JlpA protein. Taking into account that gastrointestinal tract is the focal site for C. jejuni colonization, to extrapolate the benefit of mucosal (intragastric) delivery of JlpA protein, a food grade Nisin inducible Lactic acid producing bacteria, Lactococcus lactis (L. lactis) was engineered to express recombinant JlpA protein (rJlpA) in the surface of the bacteria. Following evaluation of optimal surface expression and functionality of recombinant JlpA protein expressed by recombinant L. lactis (rL. lactis), the immune-protective role of intragastric administration of live rL. lactis was assessed in commercial broiler chickens. In addition to the significant elevation of antigen specific mucosal immune responses in the intestine of chickens that received three doses of rL. lactis, marked upregulation of Toll-like receptor 2 (TLR2) gene expression in association with mixed pro-inflammatory responses (both Th1 and Th17 type) was observed. Furthermore, intragastric delivery of rJlpA expressed by rL. lactis, but not the injectable form, resulted in a significant reduction in C. jejuni colonization in chickens suggesting that mucosal delivery of live rL. lactis expressing JlpA serves as a promising vaccine platform to induce strong immune-protective responses against C. jejuni in chickens.

Keywords: chickens, lipoprotein adhesion of Campylobacter jejuni, immuno-protection, Lactococcus lactis, mucosal delivery

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Authors: R. Majidzadeh Heravi, M. Sankian, H. Kermanshahi, M. R. Nassiri, A. Heravi Moussavi, S. A. Lari, A. R. Varasteh

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The use of probiotics engineered to express specific enzymes has been the subject of considerable attention in poultry industry because of increased nutrient availability and reduced cost of enzyme supplementation. Phytase enzyme is commonly added to poultry feed to improve digestibility and availability of phosphorus from plant sources. To construct a probiotic with potential of phytate degradation, phytase gene (appA) from E. coli was cloned and transformed into two probiotic bacteria Lactobacillus salivarius and Lactococcus lactis. L. salivarous showed plasmid instability, unable to express the gene. The expression of appA gene in L. lactis was analyzed by detecting specific RNA and zymography assay. Phytase enzyme was isolated from cellular extracts of recombinant L. lactis, showing a 46 kDa band upon the SDS-PAGE analysis. Zymogram also confirmed the phytase activity of the 46 kDa band corresponding to the enzyme. An enzyme activity of 4.9U/ml was obtained in cell extracts of L. lactis. The growth of native and recombinant L. lactis was similar in the presence of two concentrations of ox bile.

Keywords: Lactobacillus salivarus, Lactococcuslactis, recombinant, phytase, poultry

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Authors: A. Cheriguene, F. Chougrani

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A total of 153 wild lactic acid bacteria were isolated from goat’s milk collected from different areas in Western Algeria. The strains were identified using phenotypical, biochemical and physiological properties. API system and SDS-PAGE technique was also used in identification of the strains. Six genera were found Enterococcus (41.83%), Lactobacillus (29.40%), Lactococcus (19.60%), Leuconostoc (4.57%), Streptococcus thermophilus (3.26%) and Pediococcus (1.30%). The most abundant species were Enterococcus faecium (24 isolates), Enterococcus durans (22 isolates), Lactococcus lactis subsp. lactis (25 isolates), Lactobacillus rhamnosus (09 isolates) and Lactobacillus delbrueckii subsp. bulgaricus (07 isolates). The strains were screened for production and technological properties such as acid production, aminopeptidase activity, autolytic properties, antimicrobial activity and exopolysaccharide production. In general most tested isolates showed a good biomass separation when collected by centrifugation; as for the production of the lactic acid, results revealed that our strains are weakly acidifying; nevertheless, lactococci showed a best acidifying activity compared to lactobacilli. Aminopeptidase activity was also weak in most strains; but, it was generally higher for lactobacilli compared to lactococci, where we recorded 30 units for Lactobacillus delbrueckii subsp. bulgaricus M14. Autolytic activity was generally higher for most strains, more particularly lactobacilli where we recorded values of 71.13% and 70% of autolysis rate respectively in Lactobacillus rhamnosus strains 9S10 and 9S7. Antimicrobial activity was detected in 50% of the isolates, particularly in lactobacilli where 80% of strains tested were able to inhibit the growth of other strains. Two strains could produce exopolysaccharides, E. faecium 8M6 and E. durans 7S8. Some strains were able to maintain two or three technological characteristics together.

Keywords: lactic acid bacteria, technological properties, acidification, aminopeptidase acivity (AP), autolysis

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Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

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Authors: Guity Karim, Valiollah Ayareh

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Probiotic dairy products are those that contain biologically active components that may affect beneficially one or more target functions in the body, beyond their adequate nutritional effects. As far as chocolate milk is a popular dairy product in the country especially among children and youth, production of a symbiotic (probiotic + peribiotic) new product using chocolate milk, Bifidobacterium lactis (DSM, Netherland) and inulin (Bene, Belgium) would help to promote the nutritional and functional properties of this product. Bifidobacterium Lactis is used as a probiotic in a variety of foods, particularly dairy products like yogurt and as a probiotic bacterium has benefit effects on the human health. Inulin as a peribiotic agent is considered as functional food ingredient. Experimental studies have shown its use as bifidogenic agent. Chocolate milk with different percent of fat (1 and 2 percent), 6 % of sugar and 0.9 % cacao was made, sterilized (UHT) and supplemented with Bifidobacterium lactis and inulin (0.5 %) after cooling . A sample was made without inulin as a control. Bifidobacterium lactis population was enumerated at days 0, 4, 8 and 12 together with measurement of pH, acidity and viscosity of the samples. Also sensory property of the product was evaluated by a 15 panel testers. The number of live bacterial cells was maintained at the functional level of 106-108 cfu/ml after keeping for 12 days in refrigerated temperature (4°C). Coliforms were found to be absent in the products during the storage. Chocolate milk containing 1% fat and inulin has the best effect on the survival and number of B. lactis at day 8 and after that. Moreover, the addition of inulin did not affect the sensorial quality of the product. In this work, chocolate has been evaluated as a potential protective carrier for oral delivery of B. lactis and inulin.

Keywords: chocolate milk, synbiotic, bifidobacterium lactis, inulin

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Authors: Fatih Özogul, Nurten Toy, Yesim Özogul

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The influences of cell-free solutions (CFSs) of lactic acid bacteria (LAB) on cadaverine and other biogenic amine production by Listeria monocytogenes and Staphylococcus aureus were investigated in lysine decarboxylase broth (LDB) using HPLC. Cell-free solutions were prepared from Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides subsp. cremoris, Pediococcus acidophilus and Streptococcus thermophiles. Two different concentrations that were 50% and 25% CFS and the control without CFSs were prepared. Significant variations on biogenic amine production were observed in the presence of L. monocytogenes and S. aureus (P<0.05). The role of CFS on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine formation in control by L. monocytogenes and S. aureus were 500.9 and 948.1 mg/L, respectively while the CFSs of LAB induced 4-fold lower cadaverine production by L. monocytogenes and 7-fold lower cadaverine production by S. aureus. CFSs resulted in strong decreases in cadaverine and putrescine production by L. monocytogenes and S. aureus, although remarkable increases were observed for histamine, spermidine, spermine, serotonin, dopamine, tyramine, and agmatine, in the presence of LAB in lysine decarboxylase broth.

Keywords: cell-free solution, lactic acid bacteria, cadaverine, food borne-pathogen

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Authors: M. Raissy, M. Shahrani

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The present study was done to evaluate the presence of tetracycline resistance genes in Lactococcus garvieae isolated from cultured rainbow trout, West Iran. The isolates were examined for antimicrobial resistance using disc diffusion method. Of the 49 strains tested, 19 were resistant to tetracycline (38.7%), 32 to enrofloxacin (65.3%), 21 to erythromycin (42.8%), 20 to chloramphenicol and trimetoprim-sulfamethoxazole (40.8%). The strains were then characterized for their genotypic resistance profiles. The results revealed that all 49 isolates contained at least one of the tetracycline resistance genes. Tet (A) was found in 89.4% of tetracycline resistant isolates and the frequency of other gene were as follow: tet (E) 42.1%, tet (B) 47.3%, tet (D) 15.7%, tet (L) 26.3%, tet (K) 52.6%, tet (G) 36.8%, tet (34) 21%, tet (S) 63.1%, tet (C) 57.8%, tet (M) 73.6%, tet (O) 42.1%. The results revealed high levels of antibiotic resistance in L. garvieae strains which is a potential danger for trout culture as well as for public health.

Keywords: Lactococcus garvieae, tetracycline resistance genes, rainbow trout, antimicrobial resistance

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Authors: Prittesh Patel, Bhavika Patel, Ramar Krishnamurthy

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Present study was conducted to isolate lactic acid bacteria (LAB) from Oreochromis niloticus and Nemipterus japonicus fish gut. The LAB isolated were confirmed through 16s rRNA sequencing. It was observed that isolated Lactococcus spp. were able to tolerate NaCl and bile acid up to certain range. The isolated Lactococcus spp. were also able to survive in acidic and alkaline conditions. Further agro-industrial waste like peels of pineapple, orange, lemon, sugarcane, pomegranate; sweet lemon was analyzed for their polysaccharide contents and prebiotic properties. In the present study, orange peels, sweet lemon peels, and pineapple peels give maximum indigestible polysaccharide. To evaluate synbiotic effect combination of probiotic and prebiotic were analyzed under in vitro conditions. Isolates Lactococcus garvieae R3 and Lactococcus sp. R4 reported to have better fermentation efficiency with orange, sweet lemon and pineapple compare to lemon, sugarcane and pomegranate. The different agro-industrial waste evaluated in this research resulted in being a cheap and fermentable carbon source by LAB.

Keywords: agro-industrial waste, lactic acid bacteria, prebiotic, probiotic, synbiotic

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Authors: R. C. Barbuti, M. N. Oliveira, N. P. Perina, C. Haro, P. Bosch, C. S. Bogsan, J. N. Eisig, T. Navarro-Rodriguez

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Background: The management of Helicobacter pylori (H. pylori) eradication is still a matter of discussion, full effectiveness is rarely achieved and it has many adverse effects. Probiotics are believed to have a role in eradicating and possibly preventing H. pylori infection as an adjunctive treatment. The present clinical study was undertaken to see the efficacy of a specially designed fermented milk product containing Bifidobacterium lactis B420 on the eradication of H. pylori infection in a prospective, randomized, double-blind, controlled study in humans. Method: Four test products were specially designed fermented milks, counts of viable cells in all products were 1010 Log CFU. 100 mL-1 for Bifidobacterium lactis-Bifidobacterium species 420, and 1011 Log CFU. 100 mL-1 for Streptococcus thermophiles were administered to subjects infected with H. pylori with a previous diagnosis of functional dyspepsia according to the Rome III criteria in a prospective, randomized, double-blind, placebo-controlled study in humans. Results: After FM supplementation, not all subjects showed a reduction in H. pylori colonization. Conclusion: Bifidobacterium lactis B420, administered twice a day for 90 days did not show an increase in H. pylori eradication effectiveness in Brazilian patients with functional dyspepsia.

Keywords: antibacterial therapy, Bifidobacteria fermented milk, Helicobacter pylori, probiotics

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Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

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By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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Authors: Carlos A. C. G. Neto, Natan C. G. e Silva , Thaís de O. Costa, Luciana R. B. Gonçalves, Maria V. P. Rocha

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β-galactosidases (E.C. 3.2.1.23) are enzymes that have attracted by catalyzing the hydrolysis of lactose and in producing galacto-oligosaccharides by favoring transgalactosylation reactions. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers is a promising alternative to enhance the stability of the biocatalysts, among which polyethylenimine (PEI) stands out. PEI has certain properties, such as being a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases. Besides that, protects them from environmental variations. The use of chitosan support coated with PEI could improve the catalytic efficiency of β-galactosidase from Kluyveromyces lactis in the transgalactosylation reaction for the production of prebiotics, such as lactulose since this strain is more effective in the hydrolysis reaction. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from K. lactis immobilized on chitosan-coated with PEI, determining the immobilization parameters, its operational and thermal stability, and then to apply it in hydrolysis and transgalactolisation reactions to produce lactulose using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% (v/v) glutaraldehyde and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg protein per gram support. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey supplemented with fructose at a ratio of 1:2 lactose/fructose, totaling 200 g/L. Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6 with 0.1 mM MnCl2. The biocatalyst whose support was coated was named CHI_GLU_PEI_GAL, and the one that was not coated was named CHI_GLU_GAL. The coating of the support with PEI considerably improved the parameters of immobilization. The immobilization yield increased from 56.53% to 97.45%, biocatalyst activity from 38.93 U/g to 95.26 U/g and the efficiency from 3.51% to 6.0% for uncoated and coated support, respectively. The biocatalyst CHI_GLU_PEI_GAL was better than CHI_GLU_GAL in the hydrolysis of lactose and production of lactulose, converting 97.05% of lactose at 5 min of reaction and producing 7.60 g/L lactulose in the same time interval. QUI_GLU_PEI_GAL biocatalyst was stable in the hydrolysis reactions of lactose during the 10 cycles evaluated, converting 73.45% lactose even after the tenth cycle, and in the lactulose production was stable until the fifth cycle evaluated, producing 10.95 g/L lactulose. However, the thermal stability of CHI_GLU_GAL biocatalyst was superior, with a half-life time 6 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (CHI_GLU_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as, the catalytic action of the enzyme. Besides that, this process can be economically viable due to the use of an industrial residue as a substrate.

Keywords: β-galactosidase, immobilization, kluyveromyces lactis, lactulose, polyethylenimine, transgalactosylation reaction, whey

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Authors: Stephen Olusanmi Akintayo, Ilesanmi Fadahunsi

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The ability of Lactic acid bacteria (LAB) to grow and survive in milk is being exploited in industrial and biotechnological applications. Although considerable studies have been reported on the fermentation of milk, however, not so much work has been documented on the selection of LAB strains from milk of the Nigerian local cattle breeds for their starter culture potentials. A total of 110 LAB were isolated from raw milk of Sokoto gudali cattle breed. The isolates were screened for their proteolytic activities on skimmed milk media with isolates A07, F06 and A01 showing the highest zone of clearance of 18.5mm, 18.5mm, and 18.0mm respectively and were selected for the studies of their growth in different constituents of milk. A01, F06, and A07 were identified as Pediococcus acidilactici, Lactococcus raffinolactis, and Leuconostoc mesenteriodes respectively using cultural, biochemical, physiological and molecular characterization techniques. Leuconostoc mesenteriodes showed the highest growth in all the milk components that were used in this study. The three LAB species selected showed a growth range of 6.46 log cfu/ml to 10.91 log cfu/ml in lactose with Leuconostoc mesenteriodes showing the highest growth of 10.91 log cfu/ml while Pediococcus acidilactici recorded the lowest growth of 9.78 log cfu/ml. In medium containing leucine as the only amino acid, the viable counts of Pediococcus acidilactici, Lactococcus raffinolactis and Leuconostoc mesenteriodes in log cfu/ml at zero hour were 6.39, 6.36 and 6.38 respectively which increased to 9.31 log cfu/ml, 9.21 log cfu/ml, 9.92 log cfu/ml respectively after 24 hours. Similarly, in all other substrates (casein, lysine, glutamic acid, aspartic acid, stearic acid and oleic acid ) tested in this study, Leuconostoc mesenteriodes showed the highest growth. It was observed that the highest quantity of lactic acid (15.31mg/ml) was produced by Leuconostoc mesenteriodes. The same trend was also observed in the production of diacetyl and hydrogen peroxide by the three tested microorganisms. Due to its ability to grow maximally in milk components, Leuconostoc mesenteriodes shows potential as starter culture for milk fermentation.

Keywords: Leuconostoc mesenteriodes, lactic acid bacteria, Sokoto gudali, starter culture

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Authors: Carlos A. C. G. Neto, Natan C. G. Silva, Thais O. Costa, Luciana R. B. Goncalves, Maria v. P. Rocha

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Galactosidases are enzymes responsible for catalyzing lactose hydrolysis reactions and also favoring transgalactosylation reactions for the production of prebiotics, among which lactulose stands out. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers in immobilization processes is a promising alternative in order to extend the useful life of the biocatalysts, for example, the coating with polyethyleneimine (PEI). PEI is a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases and also protects it from environmental variations, for example, pH and temperature. In addition, it can substantially improve the immobilization parameters and also the efficiency of enzymatic reactions. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from Kluyveromyces lactis immobilized on PEI coated agarose, determining the immobilization parameters, its operational and thermal stability, and then to apply it in the hydrolysis of lactose and synthesis of lactulose, using whey as a substrate. This immobilization strategy was chosen in order to improve the catalytic efficiency of the enzyme in the transgalactosylation reaction for the production of prebiotics, and there are few studies with β-galactosidase from this strain. The immobilization of β-galactosidase in agarose previously functionalized with 48% (w/v) glycidol and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg/g of protein. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey (66.7 g/L of lactose) supplemented with 133.3 g/L fructose at a ratio of 1:2 (lactose/fructose). Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6, with 0.1 mM MnCl2. The biocatalysts whose supports were coated were named AGA_GLY_PEI_GAL, and those that were not coated were named AGA_GLY_GAL. The coating of the support with PEI considerably improved immobilization yield (2.6-fold), the biocatalyst activity (1.4-fold), and efficiency (2.2-fold). The biocatalyst AGA_GLY_PEI_GAL was better than AGA_GLY_GAL in hydrolysis and transgalactosylation reactions, converting 88.92% of lactose at 5 min of reaction and obtaining a residual concentration of 5.24 g/L. Besides that, it was produced 13.90 g/L lactulose in the same time interval. AGA_GLY_PEI_GAL biocatalyst was stable during the 10 cycles evaluated, converting approximately 80% of lactose and producing 10.95 g/L of lactulose even after the tenth cycle. However, the thermal stability of AGA_GLY_GAL biocatalyst was superior, with a half-life time 5 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (AGA_GLY_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as the enzyme, catalyzed reactions. In addition, the use of whey as a raw material for lactulose production has proved to be an industrially advantageous alternative.

Keywords: β-galactosidase, immobilization, lactulose, polyethylenimine, whey

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Authors: M. Kazou, A. Gavriil, O. Kalagkatsi, T. Paschos, E. Tsakalidou

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Kopanisti cheese is a Greek soft Protected Designation of Origin (PDO) cheese made of raw cow, sheep or goat milk, or mixtures of them, with similar organoleptic characteristics to that of Roquefort cheese. Traditional manufacturing of Kopanisti cheese is limited in small-scale dairies, without the addition of starter cultures. Instead, an amount of over-mature Kopanisti cheese, called Mana Kopanisti, is used to initiate ripening. Therefore, the selection of proper starter cultures and the understanding of the contribution of various microbial groups to its overall quality is crucial for the production of a high-quality final product with standardized organoleptic and physicochemical characteristics. Taking the above into account, the aim of the present study was the investigation of Kopanisti cheese microbiota and its role in cheese quality. For this purpose, four different types of Kopanisti were produced in triplicates, all with pasteurized cow milk, with the addition of (A) the typical mesophilic species Lactococcus lactis and Lactobacillus paracasei used as starters in the production of soft spread cheeses, (B) strains of Lactobacillus acidipiscis and Lactobacillus rennini previously isolated from Kopanisti and Mana Kopanisti, (C) all the species from (A) and (B) as inoculum, and finally (D) the species from (A) and Mana Kopanisti. Physicochemical and microbiological analysis was performed for milk and cheese samples during ripening. Enumeration was performed for major groups of lactic acid bacteria (LAB), total mesophilic bacteria, yeasts as well as hygiene indicator microorganisms. Bacterial isolates from all the different LAB groups, apart from enterococci, alongside yeasts isolates, were initially grouped using repetitive sequence-based polymerase chain reaction (rep-PCR) and then identified at the species level using 16S rRNA gene and internal transcribed spacer (ITS) DNA region sequencing, respectively. Sensory evaluation was also performed for final cheese samples at the end of the ripening period (35 days). Based on the results of the classical microbiological analysis, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, ranged between 7 and 10 log colony forming unit (CFU) g⁻¹, phychrotrophic bacteria, and yeast extract glucose chloramphenicol (YGC) isolates between 4 and 8 log CFU g⁻¹, while coliforms and enterococci up to 2 log CFU g⁻¹ throughout ripening in cheese samples A, C and D. In contrast, in cheese sample B, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, phychrotrophic bacteria, and YGC isolates ranged between 0 and 10 log CFU g⁻¹ and coliforms and enterococci up to 2 log CFU g⁻¹. Although the microbial counts were not that different among samples, identification of the bacterial and yeasts isolates revealed the complex microbial community structure present in each cheese sample. Differences in the physicochemical characteristics among the cheese samples were also observed, with pH ranging from 4.3 to 5.3 and moisture from 49.6 to 58.0 % in the final cheese products. Interestingly, the sensory evaluation also revealed differences among samples, with cheese sample B ranking first based on the total score. Overall, the combination of these analyses highlighted the impact of different starter cultures on the Kopanisti microbiota as well as on the physicochemical and sensory characteristics of the final product.

Keywords: Kopanisti cheese, microbiota, classical microbiological analysis, physicochemical analysis

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Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad

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With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.

Keywords: bacterial pathogen, herbal medicine, growth behavior, fish

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Authors: Chougrani Fadela, Cheriguene Abderrahim

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Fifty three strains of thermophilic and mesophilic lactic acid bacteria were isolated from the ewe’s milk. Identification reveals the presence of nineteen strains (36%) of Lactobacillus sp., seventeen strains (32%) of Lactococcus sp., nine strains (17%) of Streptococcus thermophilus and eight strains (15%) of Leuconostoc sp. The strains were characterized for their technological properties. A high diversity of properties among the studied strains was demonstrated. On the basis of technological characteristics, two strains (Lactobacillus bulgaricus and Streptococcus thermophilus) were screened with respect to their acid and flavour production for the preparation of a natural yogurt and compared to a commercial starter cultures. Sensorial analyses revealed that the product manufactured on the basis of the isolated strains have a cohesiveness and adhesiveness corresponding to standard products. The pH and the acidity recorded are also within accepted levels during all the period of conservation.

Keywords: Lactobacillus bulgaricus, Streptococcus thermophilus, yoghurt, cohesiveness, adhesiveness, Algerian ewe’s milk

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Authors: Rashad Al-Hindi

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The aims of the study were to isolate and identify potential probiotic lactic acid bacteria due to their therapeutic and food preservation importance. Sixty-three suspected lactic acid bacteria (LAB) strains were isolated from thirteen different raw milk and fermented milk product samples of various animal origins manufactured indigenously in the Kingdom of Saudi Arabia using de Man, Rogosa and Sharpe (MRS) agar medium and various incubation conditions. The identification of forty-six selected LAB strains was performed using molecular methods (16S rDNA gene sequencing). The LAB counts in certain samples were higher under microaerobic incubation conditions than under anaerobic conditions. The identified LAB belonged to the following genera: Enterococcus (16 strains), Lactobacillus (9 strains), Weissella (10 strains), Streptococcus (8 strains) and Lactococcus (3 strains), constituting 34.78%, 19.57%, 21.74%, 17.39% and 6.52% of the suspected isolates, respectively. This study noted that the raw milk and traditional fermented milk products of Saudi Arabia, especially stirred yogurt (Laban) made from camel milk, could be rich in LAB. The obtained LAB strains in this study will be tested for their probiotic potentials in another ongoing study.

Keywords: dairy, LAB, probiotic, Saudi Arabia

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Authors: Husmaini, Rijal Zein, Zulkarnain, Marlito Latifa, Syahrul E. Rambee

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The present study was conducted to evaluate the effects of probiotics–fermented purple sweet potato (PPSP) on performance and cholesterol meat of local ducks. One hundred two weeks old male local ducks placed in 4 treatment doses for ten weeks. The treatments were the dosage of PPSP, i.e., 0, 1, 2 and 3 grams of PPSP/bird/week. One gram PPSP contains 1.3 x 108 colony form unit. Data were analyzed statistically using SPSS and DMRT. The results showed that PPSP administration in local ducks did not affect intestinal villi height and fed consumption (P > 0.05), but highly significant (P < 0.01) increasing duodenum thickness, body weight, carcass yield and reducing both feed conversion and cholesterol meat content. The difference in PPSP dosage (1.2 and 3 grams) had the same effect on body weight gain. However, it has a different impact on feed conversion and meat cholesterol levels. The higher the PPSP dose given, the lower the feed conversion and meat cholesterol level. This study has shown that administration of PPSP can improve performance and reduce cholesterol levels of local duck meat. Giving PPSP as much as 3 grams per bird every week has provided the best results.

Keywords: cholesterol, local duck, performance, probiotics, purple sweet potato

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Authors: Fatih Özogul, Sezen Özçeli̇k, Yesim Özogul

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Lactic, acetic, succinic, propionic, formic and butyric acid production by lactic acid bacteria (LAB) were monitored in M17 broth (the control) and some fish (squid, shrimp, octopus, and eel) infusion broth by using HPLC method. There were significant differences in terms of lactic, acetic, succinic, propionic, formic and butyric acid production (p < 0.005) among bacterial strains. Acetic acid production was the lowest by LAB while succinic acid followed by propionic acid was synthesized at the highest levels. Lactic acid production ranged from 0 to 938 mg/L by all LAB strains in different infusion broth. The highest acetic acid production was found by Lb. acidophilus and Lb. delbrueckii subsp. lactic in octopus and shrimp infusion broth, with values of 872 and 674 mg/L, respectively while formic acid formation ranged from 1747 mg/L by Lb. acidophilus in octopus infusion broth to 69 mg/L by Lb. delbrueckii subsp. lactis in shrimp infusion broth. Propionic acid and butyric acid productions by St. thermophilus were 9852 and 3999 mg/L in shrimp infusion broth while Leu. mes. subsp. cremoris synthesized 312 and 9 mg/L of those organic acid in European squid infusion broth, respectively. Apparently, LAB strains had a great capability to generate succinic acid followed by propionic and butyric acid. In addition, other organic acid production differed significantly depending on bacterial strains and growth medium.

Keywords: Lactic acid bacteria , organic acid, HPLC analysis, growth medium

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Authors: Rashad R. Al-Hindi

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The aim of this study was to isolate and identify lactic acid bacteria and evaluate their therapeutic and food preservation importance. Ninety-three suspected lactic acid bacteria (LAB) were isolated from thirteen different raw and fermented milk of indigenous sources in the Kingdom of Saudi Arabia. The identification of forty-six selected LAB strains and genetic relatedness were performed based on 16S rDNA gene sequence comparison. The LAB counts in certain samples were higher under microaerobic than anaerobic conditions. The identified LAB belonged to genera Enterococcus (16 strains), Lactobacillus (9 strains), Weissella (10 strains), Streptococcus (8 strains) and Lactococcus (3 strains). Phylogenetic tree generated from the full-length (~1.6 kb) sequences confirmed previous findings. Utilization of shorter 16S rDNA sequences (~1.0 kb) also discriminated among strains of which V2 region was the most effective. None of the strains exhibited resistance to clinically relevant antibiotics or undesirable hemolytic activity, while they differed in other probiotic characteristics, e.g., tolerance to acidic pH, resistance to bile salt, and antibacterial activity. In conclusion, the isolates Lactobacillus casei MSJ1, Lactobacillus casei Dwan5, Lactobacillus plantarum EyLan2 and Enterococcus faecium Gail-BawZir8 are likely the best probiotic LAB and we speculate that studying the synergistic effects of bacterial combinations might result in the occurrence of more effective probiotic potential. We argue that the raw and fermented milk of animals hosted in Saudi Arabia, especially stirred yogurt (Laban) made from camel milk, are rich in LAB with promising probiotics potential.

Keywords: fermented foods, lactic acid bacteria, probiotics, Saudi Arabia

Procedia PDF Downloads 170

Authors: Thalía Moreno-García Malo, Santiago Torres-Ríos, María G. González-Cruz, María M. Hernández-Arroyo, Sergio R. Trejo-Estrada

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Agave atrovirens is the main source of agave sap, the raw material for the production of pulque, an artisanal fermented beverage, traditional since prehispanic times in the highlands of central Mexico. Agave sap is rich in glucose, sucrose and fructooligosaccharides, and strongly differs from agave syrup from A. tequilana, which is mostly a high molecular weight fructan. Agave sap is converted into pulque by a highly diverse microbial community which includes bacteria, yeast and even filamentous fungi. The bacterial diversity has been recently studied. But the composition of consortia derived from directed enrichments differs sharply from the whole fermentative consortium. Using classical microbiology methods, and selective liquid and solid media formulations, either bacterial or fungal consortia were developed and analyzed. Bacterial consortia able to catabolize specific prebiotic saccharides were selected and preserved for future developments. Different media formulations, selective for bacterial genera such as Bifidobacterium, Lactobacillus, Pediococcus, Lactococcus and Enterococcus were also used. For yeast, specific media, osmotic pressure and unique carbon sources were used as selective agents. Results show that most groups are represented in the enrichment cultures; although very few are recoverable from the whole consortium in artisanal pulque. Diversity and abundance vary among consortia. Potential bacterial probiotics obtained from agave sap and agave juices show tolerance to hydrochloric acid, as well as strong antimicrobial activity.

Keywords: Agave, pulque, microbial consortia, prebiotic activity

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Authors: Bruna Goncalves, Jennifer Henck, Romulo Uliana, Eliana Kamimura, Carlos Oliveira, Carlos Corassin

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Studies have shown evidence of human exposure to aflatoxin M1 due to the consumption of contaminated milk and dairy products (mainly cheeses). This poses a great risk to public health, since milk and milk products are frequently consumed by a portion of the population considered immunosuppressed, children and the elderly. Knowledge of the negative impacts of aflatoxins on health and economics has led to investigations of strategies to prevent their formation in food, as well as to eliminate, inactivate or reduce the bioavailability of these toxins in contaminated products This study evaluated the effect of microbiological methods using lactic acid bacteria on aflatoxin M1 (AFM1) reduction in Minas Frescal cheese (typical Brazilian product, being among the most consumed cheeses in Brazil) spiked with 1 µg/L AFM1. Inactivated lactic acid bacteria (0,5%, v/v de L. rhamnosus e L. lactis) were added during the cheese production process. Nine cheeses were produced, divided into three treatments: negative controls (without AFM1 or lactic acid bacteria), positive controls (AFM1 only), and lactic acid bacteria + AFM1. Samples of cheese were collected on days 2, 10, 20 and 30 after the date of production and submitted to composition analyses and determination of AFM1 by high-performance liquid chromatography. The reductions of AFM1 in cheese by lactic acid bacteria at the end of the trial indicate a potential application of inactivated lactic acid bacteria in reducing the bioavailability of AFM1 in Minas frescal cheese without physical-chemical and microbiological modifications during the 30-day experimental period. The authors would like to thank São Paulo Research Foundation – FAPESP (grants #2017/20081-6 and #2017/19683-1).

Keywords: aflatoxin, milk, minas frescal cheese, decontamination

Procedia PDF Downloads 157

Authors: N. Noori, P. Taherkhani, A. Akhondzadeh Basti, H. Gandomi, M. Alimohammadi

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Research on natural antimicrobial agents, especially of plant origin, highly noticed in recent years and evaluation of antimicrobial effects of native plants such as Bunium persicum Boiss. is especially important. In the present study, sensory characteristics and microbiological properties of Gouda cheese affected by different concentrations of Bunium persicum Boiss. essential oil were investigated. Extraction of the essential oil was performed by hydro distillation. The oil was analyzed by GC using flame ionization (FID) and GC/ MS for detection. The antimicrobial effects were determined against various microbial groups (aerobic mesophilic bacteria, enterococci, mesophilic lactobacilli, enterobacteriaceae, lactococcus and yeasts). Microbial groups were counted during ripening period using plate count on specific culture media. Organoleptic evaluation including teture, flavor, odor, color and total acceptability were determined at the end of aging. According to results, the essential oil yield was 4/1 % ( W/ W). Twenty- six compounds were identified in the oil that concluded 99.7 % of the total oil. The major components of Bunium persicum Boiss. essential oil were γ- terpinene- 7- al (26.9 %) and cuminaldehyde (23.3 %). Generally, the increase of Black Cumin essential oil concentration led to reduction in microbial counts in different groups. The maximum antimicrobial effect was seen in yeast that reduced by 2 log compared to the control group at EO concentration of 4µl/ ml at day 90.The minimum reduction was observed in enterobacteriaceae that showed only 0.75 log decreese compared to the control at the same concentration of EO. Addition of EO improved organoleptic properties of Gouda cheese especially in the case of flavor and odor characteristic. However, no significant differences were observed in texture and color between treatment and control groups. Bunium persicum Boiss. essential oil could be used as preservative material and flavoring agent in some kinds of food such as cheese and also could be provided consumers health.

Keywords: Bunium persicum Boiss. essential oil, Microbiological properties, sensory evaluation, gouda cheese

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Authors: M. G. Shehata, S. A. El Sohaimy, Marwa M. Abu-Serie, Nourhan M. Abd El-Aziz

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Probiotic strains can potentially be used as bio-preservatives and functional food supplement. Eight lactic acid bacteria strains (LAB) Lactobacillus brevis NRRL B-4527; Streptococcus thermophilus BLM 58; Pediococcusacidilactici ATCC 8042; Lactobacillus rhamnosus CCUG 1452; Lactobacillus curvatus ATCC 51436; Lactococcuslactis sub sp. lactisDSM 20481; Lactobacillus plantarum DMSZ 20079 and Lactobacillus plantarumTF103 were selected to screen the antioxidant, anticancer potential and probiotic properties. LAB strains exhibited good probiotic, antioxidant properties and showed antagonistic activity against food-borne pathogenic (Bacillus subtilis DB 100 host; Candida albicans ATCCMYA-2876; Clostridium botulinum ATCC 3584; Escherichia coli BA 12296; Klebsiellapneumoniae ATCC12296; Salmonella senftenberg ATCC 8400 and Staphylococcus aureus NCTC 10788). Further, in vitro probiotic properties of eight strains displayed excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, in vitro adhesion ability for HT-29 cell line. The antioxidant effect of intracellular and cell-free extract of lactic acid bacteria strains was evaluated by various antioxidant assays, namely, resistance to hydrogen peroxide, DPPH radical scavenging, ABTS radical scavenging, and hydroxyl radical scavenging (HRS). The results showed that intracellular and cell-free supernatant of S. Thermophilus BLM 58, L. lactissubsp.lactis DSM 20481, P. acidilactici ATCC 8042, L. brevis NRRL B-4527 strains possess excellent antioxidant capacity. The intracellular of S. Thermophilus BLM 58 and P. acidilactici ATCC 8042 also showed excellent anticancer activity against Caco-2, MCF-7, HepG-2, and PC-3. Antioxidative property of selected lactic acid bacteria strains would be useful in the functional food manufacturing industry. They could beneficially affect the consumer by providing dietary source of antioxidants.

Keywords: anticancer activity, antioxidant activity, functional food, lactic acid bacteria, probiotic

Procedia PDF Downloads 198

Authors: Deeba N. Baig, Ateeqa Ijaz, Saloome Rafiq

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Camel is a religious and culturally significant animal in Asian and African regions. In Pakistan Dromedary and Bactrian are common camel breeds. Other than the transportation use, it is a pivotal source of milk and meat. The quality of its milk and meat is predominantly dependent on the geographical location and variety of vegetation available for the diet. Camel milk (CM) is highly nutritious because of its reduced cholesterol and sugar contents along with enhanced minerals and vitamins level. The absence of beta-lactoglobulin (like human milk), makes CM a safer alternative for infants and children having Cow Milk Allergy (CMA). In addition to this, it has a unique probiotic profile both in raw and fermented form. Number of Lactic acid bacteria (LAB) including lactococcus, lactobacillus, enterococcus, streptococcus, weissella, pediococcus and many other bacteria have been detected. From these LAB Lactobacilli, Bifidobacterium and Enterococcus are widely used commercially for fermentation purpose. CM has high therapeutic value as its effectiveness is known against various ailments like fever, arthritis, asthma, gastritis, hepatitis, Jaundice, constipation, postpartum care of women, anti-venom, dropsy etc. It also has anti-diabetic, anti-microbial, antitumor potential along with its robust efficacy in the treatment of auto-immune disorders. Recently, the role of CM has been explored in brain-gut axis for the therapeutics of neurodevelopmental disorders. In this connection, a lot of grey area was available to explore the probiotics and therapeutics latent in the CM available in Pakistan. Thus, current study was designed to explore the predominant probiotic flora and antimicrobial potential of CM from different local breeds of Pakistan. The probiotics have been identified through biochemical, physiological and ribo-typing methods. In addition to this, bacteriocins (antimicrobial-agents) were screened through PCR-based approach. Results of this study revealed that CM from different breeds of camel depicted a number of similar probiotic candidates along with the range of limited variability. However, the nucleotide sequence analysis of selected anti-listerial bacteriocins exposed least variability. As a conclusion, the CM has sufficient probiotic availability and significant anti-microbial potential.

Keywords: bacteriocins, camel milk, probiotics potential, therapeutics

Procedia PDF Downloads 96

Authors: Natan C. G. Silva, Carlos A. C. Girao Neto, Marcele M. S. Vasconcelos, Luciana R. B. Goncalves, Maria Valderez P. Rocha

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Galactooligosaccharides (GOS) are sugars with prebiotic function that can be synthesized chemically or enzymatically, and this last one can be promoted by the action of β-galactosidases. In addition to favoring the transgalactosylation reaction to form GOS, these enzymes can also catalyze the hydrolysis of lactose. A highly studied type of GOS is lactulose because it presents therapeutic properties and is a health promoter. Among the different raw materials that can be used to produce lactulose, whey stands out as the main by-product of cheese manufacturing, and its discarded is harmful to the environment due to the residual lactose present. Therefore, its use is a promising alternative to solve this environmental problem. Thus, lactose from whey is hydrolyzed into glucose and galactose by β-galactosidases. However, in order to favor the transgalactosylation reaction, the medium must contain fructose, due this sugar reacts with galactose to produce lactulose. Then, the glucose-isomerase enzyme can be used for this purpose, since it promotes the isomerization of glucose into fructose. In this scenario, the aim of the present work was first to develop β-galactosidase biocatalysts of Kluyveromyces lactis and to apply it in the integrated reactions of hydrolysis, isomerization (with the glucose-isomerase from Streptomyces murinus) and transgalactosylation reaction, using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% glutaraldehyde was evaluated using different enzymatic loads (2, 5, 7, 10, and 12 mg/g). Subsequently, the hydrolysis and transgalactosylation reactions were studied and conducted at 50°C, 120 RPM for 20 minutes. In parallel, the isomerization of glucose into fructose was evaluated under conditions of 70°C, 750 RPM for 90 min. After, the integration of the three processes for the production of lactulose was investigated. Among the evaluated loads, 7 mg/g was chosen because the best activity of the derivative (44.3 U/g) was obtained, being this parameter determinant for the reaction stages. The other parameters of immobilization yield (87.58%) and recovered activity (46.47%) were also satisfactory compared to the other conditions. Regarding the integrated process, 94.96% of lactose was converted, achieving 37.56 g/L and 37.97 g/L of glucose and galactose, respectively. In the isomerization step, conversion of 38.40% of glucose was observed, obtaining a concentration of 12.47 g/L fructose. In the transgalactosylation reaction was produced 13.15 g/L lactulose after 5 min. However, in the integrated process, there was no formation of lactulose, but it was produced other GOS at the same time. The high galactose concentration in the medium probably favored the reaction of synthesis of these other GOS. Therefore, the integrated process proved feasible for possible production of prebiotics. In addition, this process can be economically viable due to the use of an industrial residue as a substrate, but it is necessary a more detailed investigation of the transgalactosilation reaction.

Keywords: beta-galactosidase, glucose-isomerase, galactooligosaccharides, lactulose, whey

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