Phytone

Comparison of Antibiotic-Containing Media and Media Supplemented with Dichloran and Rose Bengal for Isolation and Enumeration of Fungi in Spices.[Pubmed:30959602]

J Food Prot. 1986 Oct;49(10):815-817.

Yeast and mold counts of various spice products were determined using Dichloran Rose Bengal agar, Phytone Yeast Extract agar with added Dichloran and Rose Bengal, and Antibiotic Plate Count agar. Media containing the added Dichloran and Rose Bengal proved superior to media without Dichloran and Rose Bengal in controlling mold overgrowth, and promoting distinct colony morphology. Results were obtained 2 d earlier using Phytone Yeast Extract agar with added Dichloran and Rose Bengal.

Chemical Composition and Antimicrobial Activity of Essential Oil from Phytolacca dodecandra Collected in Ethiopia.[Pubmed:30669366]

Molecules. 2019 Jan 18;24(2). pii: molecules24020342.

The essential oil from Phytolacca dodecandra, a traditional herb of Ethiopia, has been studied, including the chemical composition and antimicrobial activity. The difference between four P. dodecandra samples (P-1(-)P-4), which differed in gender or location, has also been analyzed. The essential oils were obtained by steam distillation, while the aromas were extracted by head space solid-phase microextraction (HS-SPME) and both were analyzed by gas chromatography- mass spectrometry (GC-MS). The oils' antimicrobial activities were evaluated by the microdilution method against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Candida albicans. Ninety one components, representing 88.37 to 94.01% of the aromas, were identified. The compositions of the aromas of four samples are mainly dominated by aldehydes and ketones: 2-nonanone (1.80(-)30.80%), benzaldehyde (4.99(-)25.99%), and sulcatone (2.34(-)5.87%). Sixty components representing 64.61 to 69.64% of the oils were identified, and Phytone (3.04(-)21.23%), phytol (4.11(-)26.29%) and palmitic acid (1.49(-)23.87%) are the major compounds. No obvious antimicrobial activity was observed for all the four essential oils.

A Non-Enzymatic Pathway with Superoxide in Intracellular Terpenoid Synthesis.[Pubmed:29927025]

Angew Chem Int Ed Engl. 2018 Aug 6;57(32):10347-10351.

Non-C5 -units terpenoids (norisoprenoids) with an acetonyl group are widely distributed in nature. However, studies on the biosynthesis of norisoprenoids are scarce. Now, the C33 norisoprenoid, (all-E)-farnesylfarnesylacetone, was identified from Bacillus spp. and it was elucidated for the first time that superoxide mediates the cleavage of menaquinones (vitamin K) to form norisoprenoids in saponification treatment. From in vivo experiments using gene-disrupted Bacillus subtilis strains targeted for enzymes responsible for menaquinone biosynthesis and for superoxide dismutase, it was suggested that the non-enzymatic cleavage (autoxidation) of menaquinone with superoxide resulted in norisoprenoid synthesis in Bacillus cells. Furthermore, the bioactive norisoprenoids, farnesylacetone and Phytone, were produced in Bacillus cells by this novel synthesis system.

Suppression of the MADS-box gene SlMBP8 accelerates fruit ripening of tomato (Solanum lycopersicum).[Pubmed:28649000]

Plant Physiol Biochem. 2017 Sep;118:235-244.

MADS-box genes encode important transcription factors that are involved in many biological processes of plants, including fruit ripening. In our research, a MADS-box gene, SlMBP8, was identified, and its tissue-specific expression profiles were analysed. SlMBP8 was highly expressed in fruits of the B+4 stage, in senescent leaves and in sepals. To further characterize its function, an RNA interference (RNAi) expression vector of SlMBP8 was constructed and transferred into tomato. In the transgenic plants, the ripening of fruits was shortened by 2-4 days compared to that of wild type. At the same time, carotenoids accumulated to higher levels and the expression of Phytone synthase 1 (PSY1), phytoene desaturase (PDS) and varsigma-carotene desaturase (ZDS) was enhanced in RNAi fruits. The transgenic fruits and seedlings showed more ethylene production compared with that of the wild type. Furthermore, SlMBP8-silenced seedlings displayed shorter hypocotyls due to higher endogenous ethylene levels, suggesting that SlMBP8 may modulates the ethylene triple response negatively. A yeast two-hybrid assay indicated that SlMBP8 could interact with SlMADS-RIN. Besides, the expression of ethylene-related genes, including ACO1, ACO3, ACS2, ERF1, E4 and E8, was simultaneously up-regulated in transgenic plants. In addition, SlMBP8-silenced fruits showed higher ethylene production, suggesting that suppressed expression of SlMBP8 promotes carotenoid and ethylene biosynthesis. In addition, the fruits of transgenic plants displayed more rapid water loss and decreased storability compared to wild type, which was due to the significantly induced expressions of cell wall metabolism genes such as PG, EXP, HEX, TBG4, XTH5 and XYL. These results suggest that SlMBP8 plays an important role in fruit ripening and softening.

Production and purification of an untagged recombinant pneumococcal surface protein A (PspA4Pro) with high-purity and low endotoxin content.[Pubmed:27889801]

Appl Microbiol Biotechnol. 2017 Mar;101(6):2305-2317.

Streptococcus pneumoniae is the main cause of pneumonia, meningitis, and other conditions that kill thousands of children every year worldwide. The replacement of pneumococcal serotypes among the vaccinated population has evidenced the need for new vaccines with broader coverage and driven the research for protein-based vaccines. Pneumococcal surface protein A (PspA) protects S. pneumoniae from the bactericidal effect of human apolactoferrin and prevents complement deposition. Several studies indicate that PspA is a very promising target for novel vaccine formulations. Here we describe a production and purification process for an untagged recombinant fragment of PspA from clade 4 (PspA4Pro), which has been shown to be cross-reactive with several PspA variants. PspA4Pro was obtained using lactose as inducer in Phytone auto-induction batch or glycerol limited fed-batch in 5-L bioreactor. The purification process includes two novel steps: (i) clarification using a cationic detergent to precipitate contaminant proteins, nucleic acids, and other negatively charged molecules as the lipopolysaccharide, which is the major endotoxin; and (ii) cryoprecipitation that eliminates aggregates and contaminants, which precipitate at -20 degrees C and pH 4.0, leaving PspA4Pro in the supernatant. The final process consisted of cell rupture in a continuous high-pressure homogenizer, clarification, anion exchange chromatography, cryoprecipitation, and cation exchange chromatography. This process avoided costly tag removal steps and recovered 35.3 +/- 2.5% of PspA4Pro with 97.8 +/- 0.36% purity and reduced endotoxin concentration by >99.9%. Circular dichroism and lactoferrin binding assay showed that PspA4Pro secondary structure and biological activity were preserved after purification and remained stable in a wide range of temperatures and pH values.

Capsular polysaccharide production by Streptococcus pneumoniae serotype 1: from strain selection to fed-batch cultivation.[Pubmed:26298702]

Appl Microbiol Biotechnol. 2015 Dec;99(24):10447-56.

Streptococcus pneumoniae is a human pathogen largely transmitted by aerosols. Vaccines are the main strategy against this pathogen, and the capsular polysaccharide (PS) is its major antigen. S. pneumoniae serotype 1 is associated with large outbreaks and epidemics of invasive diseases. The aims of this work were to screen serotype 1 strains to identify the best PS1 producer, evaluate three peptones for PS1 production, investigate the effects of culture medium components using a design of experiments (DoE), a statistic tool for optimization, and propose a new medium/cultivation strategy. After flask cultivation of nine strains, two that produced high PS1 and biomass values were chosen for further evaluation in the bioreactor, and ST595/01 was chosen as the best PS1 producer strain. Among the peptones tested (Casamino acids, Soytone, and Phytone), the highest PS1 production (298 mg/L) was reached with Phytone. Next, DoE (2(4-1)) was performed to evaluate the effects of yeast extract (YE), Phytone, L-asparagine (Asn), and L-glutamine (Gln), yielding the following results: Phytone presented positive effects (p < 0.05) for maximum production of biomass, PS1, acetate, and lactate; YE showed positive effects for biomass and acid production (p < 0.05); Gln exerted a minor positive effect on PS1 yield factor on glucose (p < 0.1); and Asn presented only an effect on acetate production (p < 0.1). Hence, a new culture medium was formulated based on Phytone, YE, and glucose, and batch and fed-batch cultivations were evaluated. The fed-batch cultivation showed almost 2 times the biomass and 2.5 times the PS1 production as the batch culture, and 8-10 times higher PS1 production than has been previously reported.

Use of Phytone Peptone to Optimize Growth and Cell Density of Lactobacillus reuteri.[Pubmed:28231207]

Foods. 2015 Aug 10;4(3):318-327.

The objective of this study was to determine the use of Phytone peptone to optimize the growth and cell density of Lactobacillus reuteri. Four strains of L. reuteri (DSM 20016, SD 2112, CF 2-7F, and MF 2-3,) were used in this study. An overnight culture of individual strains was inoculated into fresh basal media with various protein sources (peptone, tryptone, proteose peptone #3, Phytone peptone, tryptic soy broth, yeast extract, and beef extract). Samples were then mixed well and incubated at 37 degrees C for 15 h. Bacterial growth was monitored by measuring turbidity (optical density 610 nm) at different time intervals during the incubation period. At the end of incubation, samples were plated on de-Man Rogosa Sharpe (MRS) agar to determine the bacterial population. Our results showed that Phytone peptone promoted the growth of L. reuteri (p < 0.05) by 1.4 log CFU/mL on average compared to the control samples. Therefore, Phytone peptone could be included in laboratory media to enhance growth and increase the cell density of L. reuteri.

Antimicrobial activity and antibiotic susceptibility of Lactobacillus and Bifidobacterium spp. intended for use as starter and probiotic cultures.[Pubmed:26019620]

Biotechnol Biotechnol Equip. 2015 Jan 2;29(1):84-91.

Antimicrobial activity and antibiotic susceptibility were tested for 23 Lactobacillus and three Bifidobacterium strains isolated from different ecological niches. Agar-well diffusion method was used to test the antagonistic effect (against Staphylococcus aureus, Escherichia coli, Bacillus cereus and Candida albicans) of acid and neutralized (pH 5.5) lyophilized concentrated supernatants (cell-free supernatant; CFS) and whey (cell-free whey fractions; CFW) from de Man-Rogosa-Sharpe/trypticase-Phytone-yeast broth and skim milk. Acid CFS and CFW showed high acidification rate-dependent bacterial inhibition; five strains were active against C. albicans. Neutralized CFS/CFW assays showed six strains active against S. aureus (L. acidophilus L-1, L. brevis 1, L. fermentum 1, B. animalis subsp. lactis L-3), E. coli (L. bulgaricus 6) or B. cereus (L. plantarum 24-4capital VE, Cyrillic). Inhibition of two pathogens with neutralized CFS (L. bulgaricus 6, L. helveticus 3, L. plantarum 24-2L, L. fermentum 1)/CFW (L. plantarum 24-5D, L. plantarum 24-4capital VE, Cyrillic) was detected. Some strains maintained activity after pH neutralization, indicating presence of active substances. The antibiotics minimum inhibitory concentrations (MICs) were determined by the Epsilometer test method. All strains were susceptible to ampicillin, gentamicin, erythromycin and tetracycline. Four lactobacilli were resistant to one antibiotic (L. rhamnosus Lio 1 to streptomycin) or two antibiotics (L. acidophilus L-1 and L. brevis 1 to kanamycin and clindamycin; L. casei L-4 to clindamycin and chloramphenicol). Vancomycin MICs > 256 mug/mL indicated intrinsic resistance for all heterofermentative lactobacilli. The antimicrobially active strains do not cause concerns about antibiotic resistance transfer and could be used as natural biopreservatives in food and therapeutic formulations.

Bifidobacterium commune sp. nov. isolated from the bumble bee gut.[Pubmed:25753540]

Antonie Van Leeuwenhoek. 2015 May;107(5):1307-13.

Bifidobacteria were isolated from the gut of Bombus lapidarius, Bombus terrestris and Bombus hypnorum bumble bees by direct isolation on modified trypticase Phytone yeast extract agar. The MALDI-TOF MS profiles of four isolates (LMG 28292(T), R-53560, R-53124, LMG 28626) were found to be identical and did not cluster with the profiles of established Bifidobacterium species. Analysis of the 16S rRNA gene sequence of strain LMG 28292(T) revealed that LMG 28292(T) is most closely related to the Bifidobacterium bohemicum type strain (96.8%), which was also isolated from bumble bee gut specimens. The hsp60 gene of strain LMG 28292(T) shows 85.8% sequence similarity to that of the B. bohemicum type strain. The (GTG)5-PCR profiles and the hsp60 sequences of all four isolates were indistinguishable; however, three different phenotypes were observed among the four isolates by means of the API 50CHL microtest system. Based on the phylogenetic, genotypic and phenotypic data, we propose to classify the four isolates within the novel species Bifidobacterium commune sp. nov., with LMG 28292(T) (= DSM 28792(T)) as the type strain.

[Chemical components from essential oil of Pandanus amaryllifolius leaves].[Pubmed:25345137]

Zhong Yao Cai. 2014 Apr;37(4):616-20.

OBJECTIVE: To analyze the chemical compositions of Pandanus amaryllifolius leaves essential oil extracted by steam distillation. METHODS: The essential oil of Pandanus amaryllifolius leaves was analyzed by gas chromatography-mass spectrum, and the relative content of each component was determined by area normalization method. RESULTS: 128 peaks were separated and 95 compounds were identified, which weighed 97.75%. The main chemical components of the essential oil were phytol (42.15%), squalene (16.81%), what's more pentadecanal (6.17%), pentadecanoic acid (4.49%), 3, 7, 11, 15-tetramethyl-2-hexadecen-1-ol (3.83%), Phytone (2.05%) and the other 74 chemical compositions were firstly identified from the essential oil of Pandanus amaryllifolius leaves. CONCLUSION: The chemical compositions of Pandanu samaryllifolius leaves essential oil was systematically, deeply isolated and identified for the first time. This experiment has provided scientific foundation for further utilization of Pandanus amaryllifolius leaves.

Effect of soy and milk protein-related compounds on Listeria monocytogenes infection in human enterocyte Caco-2 cells and A/J mice.[Pubmed:23442612]

Food Chem. 2012 Oct 15;134(4):1719-23.

Listeria monocytogenes causes listeriosis in humans, mainly through the consumption of ready-to-eat foods such as cheese. Immunocompromised persons, the elderly, and pregnant women and their fetuses or newborns are at the highest risk for the infection. We examined the effects of dietary milk-casein (MC) and soy-protein (SP), and their digested compounds tryptone (TP) and Phytone peptone (PP), respectively, on L. monocytogenes invasion and infection in human enterocyte-like Caco-2 cells and A/J mice. Invasion into Caco-2 cells tended to be high with TP. In A/J mice orally infected with L. monocytogenes, viable numbers in the liver and spleen showed a tendency of decreasing with the 20% SP diet compared to the 20% MC diet. SP suppressed the inflammation marker tumour necrosis factor-alpha in spleen tissue. Furthermore, bacteria lipopolysaccharide (LPS)-stimulated nitric oxide (NO) secretion from murine macrophage RAW 264.7 cells was suppressed by PP more than TP. These results suggest that major dietary proteins might affect infection and inflammation by L. monocytogenes.

Chemical composition and antibacterial activity of essential oils from different parts of Leonurus japonicus Houtt.[Pubmed:23344204]

Molecules. 2013 Jan 14;18(1):963-73.

The herb and fruits of Leonurus japonicus Houtt., named "Yimucao" and "Chongweizi", respectively, in Chinese, have been widely used in China as gynecological medicines. The components of the essential oils obtained by hydrodistillation were investigated by GC-MS. The antibacterial activity of the essential oils was determined by micro-dilution assay. The results showed large variations in the chemical composition and antibacterial activity of the oils. The oil of "Yimucao" showed antibacterial activity against various Gram-positive bacteria and consisted mainly of sesquiterpenes and diterpenes, with Phytone, phytol, caryophyllene oxide and beta-caryophyllene being the most significant constituents, whereas the oil of "Chongweizi", mainly made up of bornyl acetate and aliphatic hydrocarbons, was inactive in the antibacterial assay. Further study of the main compounds in "Yimucao oil" showed that beta-caryophyllene had wide-spectrum activity against Gram-positive bacteria.