Molecular structure of La3+-induced methanol dehydrogenase-like protein in Methylobacterium radiotolerans

La(3+) and not Ca(2+) increases methanol dehydrogenase (MDH) activity in Methylobacterium radiotolerans NBRC15690. La(3+)- and Ca(2+)-MDH-like proteins were found to be homodimeric (α(2)) and heterotetrameric (α(2)β(2)), respectively. N-terminal amino acid sequences of these proteins revealed that La(3+)- and Ca(2+)-MDH-like proteins were encoded by xoxF and mxaFI, respectively.     

Evaluation of Photoshop software potential for food colorimetry

This paper presents a simple method that shows how digital imaging and software analysis can be combined for color measurement. The results showed that L^∗, a^∗ and b^∗ values from Hunter colorimeter and the digital imaging method had appropriate correlation with R² of greater than 0.98, however color values obtained from digital imaging method can be used only to monitor the trend of color changes and relative comparison and there is a noticeable difference between L^∗, a^∗ and b^∗ from digital imaging and values of Hunterlab colorimeter. By using equations presented in this paper, L^∗, a^∗ and b^∗ values obtained from digital imaging method can be successfully converted to Hunterlab color values. Changes in color of Mazafati date during ripening by hot acetic acid solution were monitored by use of this method. L^∗, a^∗, and b^∗ values of the samples decreased during ripening.     

Evaluation of the performance of osmotic dehydration sheets on freshness parameters in cold-stored beef biceps femoris muscle

This study aimed to evaluate the effects of osmotic dehydration sheet (ODS) packaging on the quality parameters of beef biceps femoris muscle samples stored at 4 °C for 0, 1, 3 and 7 days. Quality indices such as Hunter color values (L^∗, a^∗ and b^∗, the percentage of metmyoglobin (Met-Mb%), K value (freshness index), and the contents of adenosine triphosphate (ATP)-related compounds (ARCs), thiobarbituric acid reactive substances (TBA-RS) and volatile basic nitrogen (VBN) were measured. ODS gave lower a^∗ and b^∗ values and lower Met-Mb% compared with control samples wrapped in polyvinylidene chloride film (PVDCF) (P < 0.01), but had no effect on L^∗ (P < 0.01). As a result, with higher levels of osmotic dehydration produced by the ODS, the percentage of weight loss and the total contents of ARCs and inosine monophosphate of the samples also increased (P < 0.05). The K values of ODS samples were also significantly lower than PVDCF-wrapped samples (P < 0.05). Low performance ODS wrapping reduced the TBA-RS values below those found with PVDCF and high performance ODS processing (P < 0.01). Moreover, the use of ODS had no effect on VBN values. Thus, although the bright red of beef samples changed to a dark purple color and the weights of samples decreased, the ODS approach has potential as a tool for decreasing the deterioration of other quality indices such as Met-Mb%, TBA-RS, ARCs, K values and the VBN content of cold-stored beef.     

Seasonal variations in chemical composition and stable isotopes of farmed and wild Brazilian freshwater fish

This study examined variations in the Fulton condition factor, chemical composition, and stable isotopes of carbon and nitrogen in the Brazilian freshwater fish cachara (Pseudoplatystoma fasciatum), comparing farmed and wild fish in different seasons. Values for energy, protein, moisture, and Fulton’s condition factor were higher for farmed than for wild fish in the rainy season, indicating better nutritional quality; however, these differences were not observed in the dry season. Likewise, we found significant enhancement of δ¹⁵N in farmed fish in the rainy season but not in the dry season, whereas enhancement of δ¹³C was observed in both seasons. The combined measurement of δ¹³C and δ¹⁵N provided traceability under all conditions. Our findings show that stable isotope analysis of C and N can be used to trace cachara origin, and that seasonal variations need to be considered when applying chemical and isotopic authentication of fish and fish products.     

SigB plays a major role in Listeria monocytogenes tolerance to bile stress

The ability of Listeria monocytogenes to tolerate high levels of bile stress is critical to its successful infection and colonization in the human gastrointestinal tract. L. monocytogenes encodes bile salt hydrolase by a bsh gene which plays a significant role in hydrolyzing high concentrations of bile salt when L. monocytogenes grows under hypoxemic condition. As the bsh promoter contains consensus SigB and PrfA binding sites, we investigated the role of SigB (σ^B) and PrfA in L. monocytogenes tolerance against bile stress by comparing the survival of isogenic deletion mutants of L. monocytogenes EGD_ΔsigB, EGD_ΔprfA and EGD_ΔprfAΔsigB with their parent strain EGD at high levels of bile salt. Our results show that the sigB deletion significantly reduced the MICs of bile salt for EGD_ΔsigB and EGD_ΔprfAΔsigB (2.6% and 2.2% vs 3.5% in wild type strain EGD), while the growth rates of these two sigB deletion mutants (EGD_ΔsigB and EGD_ΔprfAΔsigB) were affected the most in the presence of 3% bile salt. Pre-exposure to alkali (pH 9.0) and osmotic (0.3 M NaCl) stresses for a short period of time (30 min) resulted in improved growth of L. monocytogenes as well as its prfA-sigB isogenic mutants even under sublethal concentrations of bile salt, while pre-exposure to acid pH (pH 4.5) failed to provide cross-protection against subsequent bile stress. Furthermore, the sigB gene had more remarkable influence than that of prfA on bsh expression, as much lower levels of bsh transciption were observed in EGD_ΔsigB and EGD_ΔprfAΔsigB. Meanwhile, bsh expression in the deletion mutants did not respond to elevated levels of bile salt. These data indicate that σ^B might play a crucial role in Listeria survival under bile salt environment in the gastrointestinal tract before its successful colonization, invasion and intracellular propagation.     

Characterisation of phenolic acid derivatives and flavonoids from different morphological parts of Helichrysum obconicum by a RP-HPLC–DAD-(−)–ESI-MS method

The phenolic composition from different morphological parts of Helichrysum obconicum was investigated for the first time and 50 different phenolic compounds were detected. Phenolic acid conjugates, mainly mono- and di-caffeoylquinic acid derivatives, were the major components; some flavonoid derivatives were also detected in small amounts. Their separation and identification was performed by a high-performance liquid chromatography/electron spray ionisation tandem ion trap mass spectrometry method, with special emphasis on MSⁿ fragmentation. The presence of di- and tricaffeoylshikimic acid isomers in Helichrysum species extracts was reported for the first time, the spectra of these compounds were mainly characterised by the presence of a [caffeoylshikimic acid-H]⁻ ion at m/z 335. A lamiridosins-di-O-hexoside, an unusual component in Asteraceae species, was also detected.     

Molecular structure and gene analysis of Ce3+ -induced methanol dehydrogenase of Bradyrhizobium sp. MAFF211645

The molecular structure and nucleotide sequence of Ce(3+)-induced methanol dehydrogenase (MDH) of Bradyrhizobium sp. MAFF211645 were investigated. The addition of 30 μM Ce(3+) to 1/10 nutrient broth containing 0.5% methanol remarkably increased MDH activity. Furthermore, La(3+) increased MDH activity, but other heavier rare earth and metal elements did not have the same effect. MDH increased by Ce(3+) was purified by sequential column chromatography, and the purified MDH migrated as a single band with an apparent molecular weight of 68 kDa on SDS-PAGE. The apparent molecular weight of native MDH was estimated to be 108,000 by gel chromatography. The MDH was comprised of two identical subunits. N-terminal 23-amino acid sequence, 1-NDELHKMAQNPKDWVMPAGDYAN-23, of the purified MDH exhibited 91.3% identity to that of the MDH large subunit-like protein encoded by mxaF' of Bradyrhizobium japonicum USDA110. Nucleotide sequencing of the MDH gene of strain MAFF211645 yielded a deduced amino acid sequence comprising 601 amino acid residues, an N-terminal signal peptide, and a mature MDH comprising 578 amino acid residues with a predicted molecular mass of 62,918 Da. Further analysis of the deduced amino acid sequence of mature MDH revealed that the functional amino acids in its active site, such as two adjacent Cys residues, and bacterial quinoprotein signatures 1 and 2 were conserved. These results indicate that Ce(3+)-induced MDH encoded by mxaF' may be involved in methanol metabolism in Bradyrhizobium sp. MAFF211645.     

Identification and evaluation of antioxidants defatted Camellia oleifera seeds by isopropanol salting-out pretreatment

Seven flavonoid glycosides (1–7) and one theasaponin (8) were isolated from the defatted Camellia oleifera seeds using the isopropanol salting-out pretreatment (ISP) technique, followed by column chromatography. Among them, compounds 5, 7, and 8 are new compounds. The comparison of oxygen radical absorbance capacity (ORAC) of crude extracts extracted by the n-butanol partition (BP) with those extracted by the ISP showed that the latter method yields eight times greater scavenging capacities than that of the former approach. Besides, ISP exhibited almost twice the extraction efficiency than that of BP. ORAC and Trolox equivalent antioxidant capacity (TEAC) analysis revealed that the most potent antioxidants are compounds 1 and 2, which exhibited 1.78 and 1.41 times higher antioxidant activity than that of Trolox, respectively. These results indicated that the ISP approach improves the extraction efficiency in the isolation of potent antioxidants, making the defatted C. oleifera seeds valuable in the food additive industry.     

On-line separation and structural characterisation of feruloylated oligosaccharides from wheat bran using HPLC-ESI-MS

Water-soluble feruloylated oligosaccharides (FOs) from wheat bran, ferulic acid esters of arabinoxylan oligosaccharides, have been reported as natural antioxidants. In this study, structure features of FOs were studied by FT-IR spectroscopy, and high-performance liquid chromatography (HPLC) coupled to electrospray ionisation (ESI) mass spectrometry (MS) was used for separation and characterisation of FOs. Four components of FOs were resolved on a Dikma Technologies Diamonsil™ C₁₈ column eluted with water/acetonitrite (90:10) containing 0.5% HCOOH (V/V) and further analysed by ESI-MSⁿ. In ESI-MSⁿ, a predominant [M+NH₄]⁺ ion in positive mode and [M−H]⁻ ion in negative mode were observed for molecular mass information. The ESI-MSⁿ spectra of the deprotonated molecular [M−H]⁻ ion were used for structural elucidation. The structures of four isolated compounds were confirmed for the first time by on-line HPLC-ESI-MSⁿ.     

Evaluation of the effect of carvacrol on the Escherichia coli 555 metabolome by using H-NMR spectroscopy

Cultures of Escherichia coli 555 were grown at four levels of carvacrol (0–2 mM) and the E. coli endo-metabolome was extracted and measured by ¹H NMR spectroscopy. The results show that glucose concentration is going up with concentration of carvacrol and so do formate until the highest concentration is reached, from which point it suddenly decreases. This is interpreted as if the bacteria are increasingly unable to further metabolize glucose and as if the bacteria increasingly shifts with higher levels of carvacrol toward sugar fermentation as carbon source, until the level of carvacrol reaches a level (2.00 mM), where the E. coli must give up. Additionally, the multivariate Principal Component Analysis suggests that the adaptation occurring at sub-lethal doses of carvacrol is different from that occurring at higher doses.     

Activity guided isolation of antioxidants from the leaves of Ricinus communis L.

An activity-guided isolation and purification process was used to identify the DPPH (l,l-diphenyl-2-picrylhydrazyl) free radical scavenging components of the food plant (Ricinus communis L.) of Eri silkworm. Dry leaves of R. communis L. were extracted with different solvents and tested for their antioxidant activity against DPPH^•. The MeOH:water (8:2) extract showed strong DPPH radical-scavenging activity, and was subjected to column chromatography over silica gel. Gallic acid, quercetin, gentisic acid, rutin, epicatechin and ellagic acid were isolated as active components and characterised by different spectroscopic techniques.     

Analysis of isotopic ratios for the detection of illegal watering of beverages

The applications of the stable isotope ratio analyses (D/H, ¹³C and ¹⁸O) in the assessment of watering in beverages such as wine and fruit juices are discussed in this study. The measurement of ¹⁸O/¹⁶O ratio in combination with D/H and ¹³C/¹²C ratio is mainly focused and the international official methods implementing stable isotope techniques will be mentioned. Furthermore, the ongoing study on a possible use of ¹³C/¹²C ratio of CO₂ to detect the addition of technical produced CO₂ in mineral sparkling waters is also presented.     

Free radical-scavenging and H,K-ATPase inhibition activities of Pithecellobium dulce

The free radical-scavenging property and the inhibitory action on H⁺, K⁺-ATPase activity of aqueous (AEPD) and hydroalcoholic (HAEPD) fruit extracts of Pithecellobium dulce (PD) were screened in various in vitro models. HAEPD showed the greatest free radical-scavenging activity in all the experimental models. Further, HAEPD has shown a potent H⁺, K⁺-ATPase inhibitory activity and comparable to that of Omeprazole, the antiulcer drug. HPLC chromatogram of HAEPD was found to contain rich quantity of phenolic compounds and revealed the presence of flavonoids – quercitrin, rutin, kaempferol, naringin, daidzein. Thus HAEPD was found to possess a good antioxidant capacity when compared to AEPD. The study indicates that PD can be screened for antiulcer activity, since overexpression of H⁺, K⁺-ATPase results in gastric ulceration. So the free radical-scavenging and antiacid secretory activities of PD may be claimed for its traditional consumption for gastric complications.     

Implication of water activity and glass transition on the mechanical and optical properties of freeze-dried apple and banana slices

The interactions between water content, matrix structures and chemical and physical events in complex foods are usually regarded in terms of water activity and glass transition. In this work, the water content and water activity values that define important changes in the textural and optical properties of freeze-dried banana and apple slices have been related to the critical water content and water activity that cause the glass transition in both products. From the obtained results, it may be concluded that both approaches, glass transition and water activity, may be considered to prevent changes in the mechanical properties, while only the concept of water activity is needed to control the browning reactions that take place in the fruits.     

The cytochrome bcc-aa3-type respiratory chain of Rhodococcus rhodochrous

Rhodococcus rhodochrous is an active soil bacterium belonging to the Nocardia group of high GC Gram-positive bacteria. It is rich in various enzymes and thus important in the industrial production of chemicals and bioremediation. In this work, the respiratory chain of this aerobic organism was investigated and characterized. Grown under highly aerobic conditions, the membrane fraction of R. rhodochrous cells only contained a-, b- and c-type cytochromes, suggesting that it is the cytochrome bcc-aa₃-type pathway that mainly operates under these conditions. In contrast, the d-type cytochrome was also present under microaerobic conditions, indicating that the alternative pathway of the bd-type oxidase works in these circumstances. In addition, the results of H⁺/O ratio measurements indicate that these two pathways have different energy efficiencies.     

Short communication: effect of production variables on the cis-9, trans-11 conjugated linoleic acid content of cows' milk

Although there have been numerous studies investigating effects of nutrition and individual variation on the concentration of cis-9, trans-11 conjugated linoleic acid (rumenic acid; RA) in milk, there is limited information on relationships among RA content of milk and production variables. The objective of the current analysis was to examine the effects of production variables on RA content and desaturase index of milk fat. A total of 430 samples were collected from cows fed a commercial total mixed ration in winter and grazing in summer. Across a >6-fold range in production variables, RA content of milk ranged from 1 to 32 mg/g of fatty acids and desaturase index ranged from 0.03 to 0.15. Days in milk, milk yield, milk fat content, and milk fat yield had minimal or no effect on RA content of milk fat or desaturase index (R² values all <0.08). Thus, whereas nutrition and individual variation are major factors affecting RA content and desaturase index of milk fat, these values are minimally affected by days in milk, milk yield, milk fat content, and milk fat yield. Differences in these parameters do not need to be considered, therefore, when designing management strategies to increase RA content of milk fat.     

Isolation and characterization of acid-sensitive Lactobacillus plantarum with application as starter culture for Nham production

The aim of this study was to derive new starter culture variants that are unable to grow below pH 4.6, the desirable pH of the Thai fermented pork sausage, Nham, specified by Thailand Food Standard, and apply them in Nham fermentation. Several acid-sensitive mutants of one of the commercial Nham starter cultures, Lactobacillus plantarum BCC 9546, were isolated as spontaneous neomycin-resistant mutants. The growth of three representative mutants was characterized in MRS broth, which revealed that their cell numbers and acid production were lower than that of the wild-type. The H⁺-ATPase activities of the three mutants were found significantly lower than that of the wild-type under either neutral or acidic conditions. Consequently, internal pH values of the mutants appeared to be lower, especially in acidic environment (pH 5). The most acid-sensitive mutant was applied in experimental Nham production and the pH of Nham fermented with the mutant had significantly higher pH at the end of fermentation (3 days) and after an additional 4 days of storage at 30 °C. These results indicate that the use of acid-sensitive L. plantarum as starter culture can reduce the severity of post-acidification and increase the shelf life of Nham at ambient temperature.     

Isolation and evaluation of the antioxidant activity of phenolic constituents of the Garcinia brasiliensis epicarp

A new glycosylated biflavonone, morelloflavone-4′″-O-β-d-glycosyl, and the known compounds 1,3,6,7-tetrahydroxyxanthone, morelloflavone (fukugetin) and morelloflavone-7″-O-β-d-glycosyl (fukugeside) were isolated from the epicarp of Garcinia brasiliensis collected in Brazil. The structures of these compounds were established using ¹H and ¹³C NMR, COSY, gHMQC and gHMBC spectroscopy. The compounds exhibited antioxidant activity. The greatest potency was displayed by morelloflavone (2), with IC₅₀ = 49.5 mM against DPPH and absorbance of 0.583 at 400 μg/mL for the reduction of Fe³⁺. The weakest potency was displayed by 1,3,6,7-tetrahydroxyxanthone (1), with IC₅₀ = 148 mM against DPPH and absorbance of 0.194 at 400 μg/mL for the reduction of Fe³⁺.     

Evaluation of two packaging systems for regional cheese

Saloio cheese – a regional Portuguese cheese – is currently sold unpackaged or in a vacuum package. Neither of these packaging systems is acceptable: the first system yields a cheese too hard, because of excessive water loss, while the second yields a white cheese with poor textural properties. The use of a packaging system with a tailor-made moisture barrier, i.e., allowing for water loss, but at a lower rate, is a way of extending the cheese’s shelf-life.     

Selective concentration of EPA and DHA using Thermomyces lanuginosus lipase is due to fatty acid selectivity and not regioselectivity

The selectivity of anchovy oil hydrolysis was optimised for Thermomyces lanuginosus lipase, so that docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were concentrated and partially separated from each other. Enzyme concentration and pH control were important factors for effective hydrolysis. Monitoring percent hydrolysis using capillary chromatography with flame ionisation detector (Iatroscan) and fatty acid selectivity using gas chromatography (GC) indicated that during hydrolysis DHA primarily remained on the glycerol backbone, while EPA was progressively removed. ¹³C nuclear magnetic resonance (NMR) data showed that selectivity of hydrolysis was primarily due to fatty acid selectivity and not regioselectivity, with hydrolysis from both sn-1,3 and sn-2 sites being equally favoured.