Effect of medium and aggregation on antibacterial activity of nanodiamonds

Fluorescent nanodiamonds are widely used as abrasives, optical or magnetic labels, in drug delivery or nanoscale sensing. They are considered very biocompatible in mammalian cells. However, in bacteria the situation looks different and results are highly controversial. This article presents a short review of the published literature and a systematic experimental study of different strains, nanoparticle sizes and surface chemistries. Most notably, particle aggregation behaviour and bacterial clumping are taken into consideration to explain reduced colony counts, which can be wrongly interpreted as a bactericidal effect. The experiments show no mechanism can be linked to a specific material property, but prove that aggregation and bacteriostatic effect of nanodiamond attachment play a significant role in the reported results

Proline-specific peptidases from Lactobacillus casei subspecies

The objectives of this study were (l) to screen out active proline-specific peptidases from Lactobacillus casei subspecies, (2) to study growth kinetic and enzyme production from enriched medium (MRS) and cheese whey medium, (3) to purify and characterize two active proline-specific enzymes, and (4) to investigate the action of purified enzyme on bitter tryptic digests of $beta$-casein as well as bitter enzyme-modified cheese. Lactobacillus casei subsp. casei LLG and Lactobacillus casei subsp. rhamnosus S93 were examined for extra- and intra-cellular proline-specific peptidase activities. Both strains showed strong activity for x-prolyl dipeptidyl peptidase and proline iminopeptidase but had weak activities for prolidase, prolinase, and post proline endopeptidase. Histochemical staining of crude enzyme extract from Lactobacillus casei ssp. casei LLG with different substrates revealed a distinct protein band for x-prolyl dipeptidyl peptidase as well as for proline iminopeptidase. The growth kinetics showed that the intracellular proline-specific peptidases increased gradually at the beginning of the exponential phase and reached a maximum at the beginning of stationary phase.Storage stability of x-prolyl dipeptidyl peptidase and proline iminopeptidase in crude extract, with and without stabilizers showed no significant loss in activity of these two enzymes at 4$sp circ$C for 9 days without adding any stabilizers. The levels of x-prolyl dipeptidyl peptidase, proline iminopeptidase, and post proline endopeptidase activities of cells grown in whey did not vary markedly from cells grown in MRS broth. X-prolyl dipeptidyl peptidase and proline iminopeptidase were purified from crude cell-free extract of Lactobacillus casei ssp. casei LLG by Fast Protein Liquid Chromatography (FPLC) equipped with ion-exchange and gel-filtration columns. X-prolyl dipeptidyl peptidase was found to be a serine-dependent enzyme with molecular mass of 79 kDa. The pH and the temperature optima by the purified enzyme were 7.0 and 50$sp circ$C, respectively. Proline iminopeptidase was sulfhydryl enzyme with molecular mass of 46 kDa. The maximum enzyme activity was observed at pH 7.5 and 40$sp circ$C. This is the first report describing the purification and characterization of x-prolyl dipeptidyl peptidase and proline iminopeptidase from Lactobacillus casei to homogeneity.The debittering of tryptic digests from $beta$-casein by x-prolyl dipeptidyl peptidase was studied by reversed phase high performance liquid chromatography (RP-HPLC) and liquid chromatography/mass spectrometry. The results showed that two bitter peptides (f53-97 and f03-209) containing X-Pro-Y-Pro in their amino acid residues were completely hydrolyzed and many other peptides with high hydrophobicity were decreased in peak area. The addition of purified x-prolyl dipeptidyl peptidase on bitter enzyme-modified cheese (EMC) also showed that at least one bitter peptide with X-Pro-Y derived from $alpha$-casein hydrolysis was removed

Production and evaluation of enzyme-modified lighvan cheese using different levels of commercial enzymes

Enzyme-modified cheeses are concentrated cheese flavors produced enzymatically from dairy substrates in order to provide an intense source of cheese flavor with broad applications. Lighvan cheese is an Iranian traditional cheese with a pleasant taste and flavor generated after ripening. Therefore, the objective of the present study was to use commercial enzymes to produce enzyme-modified Lighvan cheese made from unripened and immature cheese. In this study, Neutrase (0.05%, 0.15%, and 0.2%) and Flavourzyme (0.05%, 0.1%, and 0.2%) were added to the base mixture. The resulting mixture was stored in an incubator for 24, 72, and 96 h to provide intense cheese flavor. Sensory evaluations of all samples in terms of bitterness, flavor, taste, and general acceptance were also carried out. The results of the sensory evaluations revealed no significant difference between most of the samples in terms of bitterness, flavor, taste, and general acceptance with respect to the incubation duration and the type and level of the commercial enzymes (p ≤ 0.05). However, the effect of the different concentrations of Flavourzyme on the cheese texture was significant after 24, 72, and 96 h of incubation (p ≤ 0.05). In addition, the effects of the different concentrations of Neutrase on the cheese texture were significant after 96 h of incubation (p ≤ 0.05). Finally, the effect of different concentrations of Flavourzyme on the general acceptance of the samples was significant following 24, 72, and 96 h of incubation (p ≤ 0.05). In general, considering the flavor, taste, texture and general acceptance scores of the enzyme-modified Lighvan cheese samples, the best sample was the sample produced by using 0.1% Neutrase and 0.1% Flavourzyme mixture

The biodiversity of Lactobacillus spp. from Iranian raw milk Motal cheese and antibacterial evaluation based on bacteriocin-encoding genes

Abstract Lactobacilli, as the largest group of lactic acid bacteria, produce large amounts of antimicrobial metabolites such as organic acids, fatty acids, ammonia, hydrogen peroxide, diacetyl and bacteriocin, which inhibit the growth of pathogenic bacteria and increase shelf life of food. The aim of this study was to identify the Lactobacillus spp. isolated from Iranian raw milk Motal cheese and to detect the presence of bacteriocin genes in the isolated Lactobacillus strains exhibiting antimicrobial activity. For this purpose, 6 Motal cheese samples from Dasht-e-Moghan region, Iran, were subjected to microbial characterization. Nineteen Lactobacillus spp. were isolated and subsequently identified based on biochemical and molecular methods. According to the sequencing of isolates, Lactobacillus spp. consisted primarily of Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei and Lactobacillus buchneri. The identified isolates were then evaluated for antimicrobial activity against Escherichia coli ATCC 25922, Listeria innocua ATCC 33090 and Staphylococcus aureus ATCC 25923. The results of PCR analysis using specific primers of genes encoding Bacteriocin, revealed the presence of Plantaricin A and Plantaricin EF in all Lactobacillus plantarum isolates and Brevicin 174A in 5 of Lactobacillus brevis isolates, whereas the gene encoding Pediocin PA-1 was not observed in any of examined isolates. It is therefore concluded that bacteriocinogenic isolates could be recommended as suitable candidates to be used as starter, adjunct-starter or antimicrobial agents for production of fermented and non-fermented products

Effect of the milk fat content and starter culture selection on proteolysis and antioxidant activity of probiotic yogurt

In this study, the effects of milk fat content (0%, 2% and 3.5%) and starter culture (autochthonous or commercial) on physicochemical properties, degree of proteolysis, antioxidant activity and viability of Lactobacillus acidophilus, within 21 days storage of probiotic yogurt at 5 ± 1 °C were investigated. Statistical analysis showed that the type of starter culture had a significant effect (P < 0.05) on proteolysis and antioxidant activity, in such a way that both of them were increased until the 14th day of storage but they decreased after this period. Similarly, the pH value of all samples decreased during storage time. It ranged from 3.84-4.34 and 4.18–4.43 for yogurt samples made by autochthonous and commercial starter culture, respectively. According to the results, the survival of Lactobacillus acidophilus decreased during storage time (P < 0.05), although it stood at recommended levels for health effects (at least 106 cfu/ml in traditional yogurt). Milk fat content did not have significant effect on the survival of probiotic organisms (P < 0/05)

Interactions between polyols and wheat biopolymers in a bread model system fortified with inulin: A Fourier transform infrared study

One of the ways to improve food safety and reduce community health risks is fortification of these products with inulin. Inulin, in spite of the effects and nutritional benefits, will also have undesirable effects on the quality and shelf life of bread. In this study, the interactions between polyols as improvers (i.e. glycerol, sorbitol and propylene glycol) and major biopolymers of wheat flour (i.e. starch and gluten) were examined in model systems fortified with Serish inulin by Fourier transform infrared (FTIR) spectroscopy. The changes in starch structure were estimated focusing on the ratios of the heights of the bands at 1047 and 1022 cm−1 which expresses the quantity of ordered starch to amorphous starch. At first and 5th days of storage, this ratio of control sample was higher than polyol treated samples. It was proved from Gaussian–Lorenzian curve fitting that the relative contribution of characteristic peaks of β-turns and intramolecular β-sheets was consecutively increased when polyol proportion of models increased. Whereas, content of intermolecular β-sheets and α-helix was slightly decreased with increasing of polyols in the models. Briefly, polyols especially 5% propylene glycol, could be used to reduce the undesirable effects of inulin on the quality parameters of dough and bread

The Emulsifier Carboxymethylcellulose Induces More Aggressive Colitis in Humanized Mice with Inflammatory Bowel Disease Microbiota Than Polysorbate-80

Commonly used synthetic dietary emulsifiers, including carboxymethylcellulose (CMC) and polysorbate-80 (P80), promote intestinal inflammation. We compared abilities of CMC vs. P80 to potentiate colitis and impact human microbiota in an inflammatory environment using a novel colitis model of ex-germ-free (GF) IL10−/− mice colonized by pooled fecal transplant from three patients with active inflammatory bowel diseases. After three days, mice received 1% CMC or P80 in drinking water or water alone for four weeks. Inflammation was quantified by serial fecal lipocalin 2 (Lcn-2) and after four weeks by blinded colonic histologic scores and colonic inflammatory cytokine gene expression. Microbiota profiles in cecal contents were determined by shotgun metagenomic sequencing. CMC treatment significantly increased fecal Lcn-2 levels compared to P80 and water treatment by one week and throughout the experiment. Likewise, CMC treatment increased histologic inflammatory scores and colonic inflammatory cytokine gene expression compared with P80 and water controls. The two emulsifiers differentially affected specific intestinal microbiota. CMC did not impact bacterial composition but significantly decreased Caudoviricetes (bacteriophages), while P80 exposure non-significantly increased the abundance of both Actinobacteria and Proteobacteria. Commonly used dietary emulsifiers have different abilities to induce colitis in humanized mice. CMC promotes more aggressive inflammation without changing bacterial composition