Potential molecular mechanism of reuterin on the inhibition of Aspergillus flavus conidial germination: An in silico study.

Reuterin is a natural antifungal agent derived from certain strains of Limosilactobacillus reuteri. Our previous study revealed that 6 mM reuterin inhibited completely the conidial germination of aflatoxigenic Aspergillus flavus. This study investigated the potential molecular mechanism of reuterin in inhibiting A. flavus conidial germination, which was pre-assumed that it correlated to the inhibition of some essential enzyme activity involved in conidial germination, specifically 1,3-ß-glucan synthase, chitin synthase, and catalases (catalase, bifunctional catalase-peroxidase, and spore-specific catalase). The complex of 1,3-ß-glucan synthase and chitin synthase with reuterin had a lower binding affinity than that with the substrate. Conversely, the complex of catalases with reuterin had a higher binding affinity than that with the substrate. It was suggested that 1,3-ß-glucan synthase and chitin synthase tended to bind the substrate rather than bind reuterin. In contrast, catalases tended to bind reuterin rather than bind the substrate. Therefore, reuterin could be a potential inhibitor of catalases but may not be an inhibitor of 1,3-ß-glucan synthase and chitin synthase. In this in silico study, we predicted that the potential molecular mechanism of reuterin in inhibiting A. flavus conidial germination was due to the inhibition of catalases activities by competitively binding to the enzymes active sites, thus resulting in the accumulation of reactive oxygen species in cells, leading to cells damage. PRACTICAL APPLICATION: This in silico study revealed that reuterin is a potential inhibitor of catalases in A. flavus, thereby interfering with the antioxidant system during conidial germination. This finding shows that reuterin can be used as an antifungal agent in food or agricultural products, inhibiting conidial germination completely.

The Occurrence of Folate Biosynthesis Genes in Lactic Acid Bacteria from Different Sources.

Research background: Lactic acid bacteria (LAB) are known to produce folate. However, this ability is highly strain-dependent. Folate synthesis in specific LAB strains is affected by the availability of folate, which can be consumed by other LAB under certain conditions. Moreover, differences in folate synthesis capabilities are related to the presence of folate biosynthesis-related genes and regulation of this pathway. Experimental approach: As basic information to better understand the regulation of folate biosynthesis among different LAB species and strains, folate biosynthetic genes were screened and identified in folate-producing and non-folate-producing LAB isolated from various local food sources in Indonesia. The extracellular folate productivity amounts of the isolates were analyzed using high-performance liquid chromatography with a diode array detector (HPLC-DAD). Results and conclusions: Eleven of the thirteen tested LAB isolates had all of the eight genes involved in folate biosynthesis (folE, folQ, folB, folK, folP, folC1, folA and folC2). Furthermore, these isolates produced extracellular folate ranging from 10.37 to 31.10 µg/mL. In contrast, two non-folate-producing isolates lacked several folate biosynthetic genes, such as folQ, folP and folA, which is possibly the reason for their inability to synthesize folate de novo. Phylogenetic tree construction revealed that the folate biosynthetic genes (excluding folK and folP) from six distinct species of folate-producing LAB isolates were monophyletic with homologous genes from other LAB species in the database. Novelty and scientific contribution: In this study, the distribution of folate biosynthetic genes in various LAB species was determined. The findings from this research support the use of folate biosynthesis marker genes in the genotypic screening for folate-producing LAB.

Effect of processing treatments on the allergenicity of nuts and legumes: A meta-analysis.

The effective food processing to reduce nuts and legumes allergenicity could not be easily and directly concluded from reading a few published reports. Therefore, we conducted a meta-analysis to investigate this issue. A literature search was conducted in eight electronic databases from January 2000 to June 11, 2021. The primary outcome of interest was the allergenicity of processed nuts or legumes determined by enzyme-linked immunosorbent assay from in vitro studies. Data with the standardized mean difference (SMD) of 95% confidence interval (CI) were pooled using a random-effect model by RevMan 5.4 software. Heterogeneity was assessed using Cochran's Q (PQ ) and I2 tests. The search strategy identified 18,793 articles. However, only 61 studies met the inclusion criteria and were included in this meta-analysis. There were 21 and 15 types of respective single and combined food processing treatments analyzed for their effects on reducing allergenicity. In single processing treatment, the extrusion and fermentation had the largest reduction in allergenicity, considering their SMD value, that is, -20.19 (95% CI: -22.22 to -18.17; the certainty of evidence: moderate) and -20.8 (95% CI: -24.10 to -17.50; the certainty of evidence: moderate), respectively. Whereas in the combination, the treatment of fermentation followed by proteolytic hydrolysis showed the most significant reduction (SMD: -53.34; 95% CI: -70.18 to -36.5) and the evidence quality of this treatment was considered moderate. In conclusion, these three food processing methods showed a desirable impact in reducing nuts or legumes allergenicity. PRACTICAL APPLICATION: Nuts and legumes play an essential role as protein sources in food consumption worldwide, but they usually contain allergens. Our study has investigated the food processing methods that effectively reduce their allergenicity by meta-analysis. The result gives valuable information for further laboratory investigation on allergens and can be used by food industries in providing foods from nuts and legumes with lower allergenicity.

Hypothetical Regulation of Folate Biosynthesis and Strategies for Folate Overproduction in Lactic Acid Bacteria.

Folate (vitamin B9) is an essential nutrient for cell metabolism, especially in pregnant women; however, folate deficiency is a major global health issue. To address this issue, folate-rich fermented foods have been used as alternative sources of natural folate. Lactic acid bacteria (LAB), which are commonly involved in food fermentation, can synthesize and excrete folate into the medium, thereby increasing folate levels. However, screening for folate-producing LAB strains is necessary because this ability is highly dependent on the bacterial strain. Some strains of LAB consume folate, and their presence in a fermentation mix can lower the folate levels of the final product. Since microorganisms efficiently regulate folate biosynthesis to meet their growth needs, some strains of folate-producing LAB can deplete folate levels if folate is available in the media. Such folate-efficient producers possess a feedback inhibition mechanism that downregulates folate biosynthesis. Therefore, the application of folate-overproducing strains may be a key strategy for increasing folate levels in media with or without available folate. Many studies have been conducted to screen folate-producing bacteria, but very few have focused on the identification of overproducers. This is probably because of the limited understanding of the regulation of folate biosynthesis in LAB. In this review, we discuss the roles of folate-biosynthetic genes and their contributions to the ability of LAB to synthesize and regulate folate. In addition, we present various hypotheses regarding the regulation of the feedback inhibition mechanism of folate-biosynthetic enzymes and discuss strategies for obtaining folate-overproducing LAB strains.

Purification, HR-LC-ESI-MS-MS Identification, and Peptide Prediction of Bacteriocin-Like Inhibitory Substances Produced by Streptomyces sp. Isolated from Chanos chanos.

Consumption of fresh and minimally processed food is closely related to foodborne diseases. To minimize the adverse effects, bacteriocin-like inhibitory substance (BLIS) as a natural preservative can be used. One of the bacteriocins with promising activity was produced by Streptomyces sp. Using gel filtration chromatography, the bacteriocin purification process succeeded in obtaining semi-purified fractions with broad-spectrum inhibitory activity to foodborne pathogen bacteria. These fractions are also stable up to 100 °C and pH 2.0-7.0. High-Resolution Liquid Chromatography Electrospray Ionization-Tandem Mass Spectrometry analysis followed by orthogonal projection to latent structure showed that each fraction had eight peaks with the highest positive correlation to BLIS-specific activity. Peptide identification based on MS spectrum found 597 predictive peptides, of which 42 predictive peptides with antimicrobial peptide characteristics and the highest iAMPpred antimicrobial peptide probability (>0.5) were selected. The selected predictive peptides have molecular mass of 247.13-615.37 Da and consist of at least 20% hydrophobic amino acids with a hydrophobicity value of 14.72 Kcal mol-1. The results of this study indicate the effectiveness of BLIS purification by gel filtration chromatography and the promising potential of semi-purified BLIS as a natural preservative. Besides, the active peptides in semi-purified BLIS can also be identified quickly so that the isolation process to obtain purified-BLIS can be carried out more efficiently.

Thermal Stability of Anisoyl Kaempferol Glycosides in Jack Bean (Canavalia ensiformis (L.) DC) and Their Effect on α-Glucosidase Inhibition.

Kaempferol glycosides are functional components of jack bean. The chemical stability of kaempferol glycosides under simulated food processing conditions was evaluated in this study by subjecting the methanol extract and each compound to heat treatment. During the heat treatment, rearrangement of the anisoyl group on the rhamnose moiety of the kaempferol glycoside was observed, followed by hydrolysis upon long-term heat treatment. One of the two regioisomers produced under heating conditions showed higher α-glucosidase inhibitory activity than the dominant anisoyl kaempferol glycoside. This rearrangement reaction was also observed upon the heat treatment of methyl-3-O-anisoyl-rhamnose, with the rearrangement from the 3-position to the 2-position occurring preferentially. The approach adopted in this study can be used to design appropriate food processing conditions, which, in turn, will increase the functional value of foods.

The Bioactivity Prediction of Peptides from Tuna Skin Collagen Using Integrated Method Combining In Vitro and In Silico.

The hydrolysates and peptide fractions of bigeye tuna (Thunnus obesus) skin collagen have been successfully studied. The hydrolysates (HPA, HPN, HPS, HBA, HBN, HBS) were the result of the hydrolysis of collagen using alcalase, neutrase, and savinase. The peptide fractions (PPA, PPN, PPS, PBA, PBN, PBS) were the fractions obtained following ultrafiltration of the hydrolysates. The antioxidant activities of the hydrolysates and peptide fractions were studied using the DPPH method. The effects of collagen types, enzymes, and molecular sizes on the antioxidant activities were analyzed using profile plots analysis. The amino acid sequences of the peptides in the fraction with the highest antioxidant activity were analyzed using LC-MS/MS. Finally, their bioactivity and characteristics were studied using in silico analysis. The hydrolysates and peptide fractions provided antioxidant activity (6.17-135.40 µmol AAE/g protein). The lower molecular weight fraction had higher antioxidant activity. Collagen from pepsin treatment produced higher activity than that of bromelain treatment. The fraction from collagen hydrolysates by savinase treatment had the highest activity compared to neutrase and alcalase treatments. The peptides in the PBN and PPS fractions of <3 kDa had antidiabetic, antihypertensive and antioxidant activities. In conclusion, they have the potential to be used in food and health applications.

Folate in Milk Fermented by Lactic Acid Bacteria from Different Food Sources.

Folates are essential micronutrients, and folate deficiency still occurs in many countries. Lactic acid bacteria (LAB) are known to be able to synthesize folates during fermentation, but the folate production is strain-dependent and influenced by the fermentation medium, presence of a folate precursor, and fermentation time. This study aimed to screen extracellular folate-producing LAB from local food sources and evaluate the factors influencing their folate biosynthesis during milk fermentation. The selection of folate-producing LAB was based on their ability to grow in folate-free medium (FACM), with folate concentrations quantified by microbiological assay. Growth of the 18 LAB in FACM varied between isolates, with only 8 isolates growing well and able to synthesize extracellular folate at relatively high concentrations (up to 24.27 ng/mL). The isolates with highest extracellular folate levels, Lactobacillus fermentum JK13 from kefir granules, Lactobacillus plantarum 4C261 from salted mustard, and Lactobacillus rhamnosus R23 from breast milk, were applied to milk fermentation. The last two isolates were probiotic candidates. The three isolates consumed folate when it was present in the milk, and its consumption was in line with their growth. The availability of folate precursors affected the amount of folate consumed, but did not lead to increased folate concentrations in the medium after 72 h fermentation. The results of this study indicate that these isolates cannot be utilized for producing folate in folate-containing milk, as it shows feedback inhibition on folate biosynthesis.

Chemical and Antioxidant Characteristics of Skin-Derived Collagen Obtained by Acid-Enzymatic Hydrolysis of Bigeye Tuna (Thunnus obesus).

The utilization of bigeye tuna skin as a source of collagen has been increasing the value of these skins. In this study, the quality of the skin was studied first. The skin after 14 h freeze-drying showed a high protein level (65.42% ± 0.06%, db), no histamine and a lack of heavy metals. The collagens were extracted through acid and acid-enzymatic methods. The enzymes used were bromelain, papain, pepsin, and trypsin. The two highest-yield collagens were pepsin-soluble collagen (PSC) and bromelain-soluble collagen (BSC). Both were type I collagen, based on SDS-PAGE and FTIR analysis. They dissolved very well in dimethyl sulfoxide and distilled water. The pH ranges were 4.60-4.70 and 4.30-4.40 for PSC and BSC, respectively. PSC and BSC were free from As, Cd, Co, Cr, Cu, and Pb. They showed antioxidant activities, as determined by the DPPH method and the reducing power method. In conclusion, bigeye tuna skin shows good potential as an alternative source of mammalian collagen. Although further work is still required, PSC and BSC showed the potential to be further used as antioxidant compounds in food applications. Other biological tests of these collagens might also lead to other health applications.

Umami fractions obtained from water-soluble extracts of red oncom and black oncom-Indonesian fermented soybean and peanut products.

Red oncom, a fermented product from solid waste of soybean curd process, and black oncom, a similar fermented product but made from defatted peanut cake, have been known to have umami taste. Umami fractions of red oncom and black oncom that are responsible for umami taste have not been investigated yet. The objective of this research was to characterize umami fractions obtained by ultrafiltration and chromatography of both oncoms. The first step, water-soluble extracts of oncoms were ultrafiltered using a membrane with cutoff 3,000 Da and followed by gel filtration chromatography (Sephadex G-25) to obtain umami fractions. Ultrafiltration fractions of red oncom (molecular weight [MW] less than 3,000 Da) and black oncom (MW more than 3,000 Da) had an intense umami taste. The further fractionation by gel filtration chromatography linked to taste dilution analysis yielded umami fractions. Chemical characterization revealed that free glutamic acid, free phenylalanine, and peptides containing their residual amino acids were present in the fractions. PRACTICAL APPLICATION: Umami fractions of red and black oncoms can be used as a source of umami compounds for food industries and food services. The information from this paper can be used by other researchers who will explore umami peptides.

Antidiabetic components from the hexane extract of red kidney beans (Phaseolus vulgaris L.): isolation and structure determination.

Red kidney beans (Phaseolus vulgaris L.) contain bioactive compounds that are known to exhibit antidiabetic effects via inhibition of α-glucosidase. However, information on the nonpolar components that exhibit antidiabetic activity is limited. Here, we report the isolation and structure determination of components with α-glucosidase inhibitory activity, which were obtained from the hexane extract of red kidney beans. Triacylglycerols (TAGs) were identified as the major components exhibiting inhibitory activity against α-glucosidase. The chemical structure of TAGs was determined by a combination of GC-MS and UPLC-MS/MS. The primary TAGs identified were LnLnLn (trilinolenin) and LnLLn (1,3-dilinolenoyl-2-linoleoyl glycerol). The major fatty acids present in these TAGs were α-linolenic acid (ω-3) and linoleic acid (ω-6). These TAGs were also found to inhibit the α-glucosidase activity in a similar fashion as acarbose. These results suggest that TAGs have potency as antidiabetics and support the potential suitability of red kidney beans for diabetes treatment.

The proportion-ratio on dipeptidyl aminopeptidase-4 (DP-4) inhibition by gelatin compared to synthetic sitagliptin.

The current study aims to determine the inhibition activity gelatin against dipeptidyl aminopeptidase 4 (DP-4). Two commercial gelatins, i.e., bovine and fish skin gelatin and one extracted (in our laboratory) gelatin, i.e., fish bone gelatin were selected for analysis. Each gelatin have same protein pattern (75-245 kDa) on sodium dodecyl sulfate polyacrylamide gel electrophoresis with mean of protein concentration of 1.72 mg/mL. The inhibition activity was measured on the capacity to inhibit DP-4 by using Gly-Pro-p-nitroanilide as their substrate. The sitagliptin was used as standard comparison. Based on the percent inhibition, gelatin has been shown to be the prospective DP-4 inhibitor.

Proteomic study of bioactive peptides from tempe.

Tempe is a traditional Indonesian fermented soybean mostly produced in small industries and sold locally throughout the country. Studies on the bioactive peptides in tempe are rare. Here, we studied bioactive peptides in samples from three tempe producers with different degrees of sanitation. The peptide sub-fractions of tempe from each producer were collected following water extraction, ultrafiltration (<3 kDa), gel filtration chromatography, and reversed phase-high performance liquid chromatography (RP-HPLC) separation followed by liquid chromatography-mass spectrometry (LC-MS). The MS spectra were then predicted using FindPept tools, and their biofunctionalities were confirmed with BIOPEP databases. There were few similar peptides found in tempe from the three producers. Peptides Val-His and Ala-Leu-Glu-Pro were found in tempe from all producers. Producers having a good sanitation level had more bioactive peptides than those with moderate or poor sanitation levels (58%, 43% and 35%, from good to poor sanitation). This work showed that the tempe from the three producers had antihypertensive, antidiabetic, antioxidative and antitumor peptides.

Umami compounds present in low molecular umami fractions of asam sunti - A fermented fruit of Averrhoa bilimbi L.

Salted fermented fruit known as asam sunti (Averrhoa bilimbi L.) in Indonesia has been used as a source of umami taste. This study was aimed to characterize the three types of asam sunti and their water soluble extracts, and to trace the compounds responsible for umami taste in umami fractions. Umami fractions were obtained by ultrafiltration followed by Sephadex G-15 chromatographic separation. The three types of samples could be differentiated by physicochemical and sensory analyses. Low molecular weight fraction had the highest umami intensity. Further chromatographic separation revealed three umami fractions, F-II, III and IV. Umami taste of F-III was due to the presence of free l-glutamic acid at 6 times, while FII and FIV were due to succinic acid at more than 30 times their respective umami thresholds. Organic acid as well as amino acid seemed to play an important role in the intense umami taste of asam sunti.

Taste and chemical characteristics of low molecular weight fractions from tofuyo - Japanese fermented soybean curd.

Tofuyo, a Japanese traditional food, is a fermented soybean curd manufactured in Okinawa region. Due to its original cheese-like flavor, the current study was designed to evaluate the sensory and chemical characteristics of three stepwise ultrafiltration fractions, using 10,000, 3000 and 500 Da membranes and further chromatographic fractions from tofuyo. The results showed that umami, sweet and salty were the characteristic tastes of all fractions, with umami intensity evaluated for the fraction with MW less than 500 Da (F-500) as the most prominent among the three fractions. Subsequent Sephadex G-25 SF fractions and RP-HPLC fractions were subjected to sensory and chemical analyses. The tastiest fraction contained sodium chloride, sugars, organic acids, umami and sweet free amino acids, at concentrations above their thresholds. The abundant presence of umami and sweet free amino acids with certain concentrations of sodium chloride and glucose might provide the typical savory taste of tofuyo.

Accumulation patterns of lipophilic organic contaminants in surface sediments and in economic important mussel and fish species from Jakarta Bay, Indonesia.

Non-target screening analyses were conducted in order to identify a wide range of organic contaminants in sediment and animal tissue samples from Jakarta Bay. High concentrations of di-iso-propylnaphthalenes (DIPNs), linear alkylbenzenes (LABs) and polycyclic aromatic hydrocarbons (PAHs) were detected in all samples, whereas phenylmethoxynaphthalene (PMN), DDT and DDT metabolites (DDX) were detected at lower concentrations. In order to evaluate the uptake and accumulation by economic important mussel (Perna viridis) and fish species, contaminant patterns of DIPNs, LABs and PAHs in different compartments were compared. Different patterns of these contaminant groups were found in sediment and animal tissue samples, suggesting compound-specific accumulation and metabolism processes. Significantly higher concentrations of these three contaminant groups in mussel tissue as compared to fish tissue from Jakarta Bay were found. Because P. viridis is an important aquaculture species in Asia, this result is relevant for food safety.

Soy sauce and its umami taste: a link from the past to current situation.

Soy sauce taste has become a focus of umami taste research. Umami taste is a 5th basic taste, which is associated to a palatable and pleasurable taste of food. Soy sauce has been used as an umami seasoning since the ancient time in Asia. The complex fermentation process occurred to soy beans, as the raw material in the soy sauce production, gives a distinct delicious taste. The recent investigation on Japanese and Indonesian soy sauces revealed that this taste is primarily due to umami components which have molecular weights lower than 500 Da. Free amino acids are the low molecular compounds that have an important role to the taste, in the presence of sodium salt. The intense umami taste found in the soy sauces may also be a result from the interaction between umami components and other tastants. Small peptides are also present, but have very low, almost undetected umami taste intensities investigated in their fractions.