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A novel antibacterial peptide specific to Streptococcus pyogenes was produced from dried fruit protein of Brucea javanica (L.) Merr. A mixture of active peptides from the fruit protein was produced in vitro by pepsin hydrolysis. The hydrolysate was purified by reverse-phase HPLC, and antimicrobial peptides active against Gram-negative...
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... on this procedure, we obtained protein hydrolysate with a maximum hydrolysis of 3Á7 AE 0Á88% (n = 3). Antimicrobial activity of the protein hydrolysate by agar dilution analysis indicated a broad-spectrum activity with the most potent growth inhibition against Gram-negative bacteria: Acinetobacter calcoaceticus (ATCC 23055), Pseudomonas fluorescens (DMST 6034) and Gram- positive bacteria: Streptococcus pyogenes (DMST 17020) at a MIC value of 7Á5 lg protein ml À1 ( Table 1). The antibac- terial activity was lower when tested against Acinetobacter baumannii (ATCC 19066), Acinetobacter lwoffii (ATCC 15309), Vibrio cholerae non-O1, non-O139 (DMST 2873) and Serratia marcescens (ATCC 8100) at a MIC value of 15 lg protein ml À1 . ...
Citations
... The proteins were hydrolyzed using proteinase K and pepsin as described previously by Jeenkeawpieam et al. [22], Jeenkeawpieam et al. [24] and Sornwatana et al. [26] with slight modifications. The proteins were incubated at a 1:25 enzyme-to-protein ratio with porcine pepsin (P7000-100G, 250 units/mg) (Sigma-Aldrich, Missouri, USA) or proteinase K (Invitrogen, Thermo Fisher Scientific ™ , California, USA). ...
Background and Aim
Antimicrobial resistance is an emerging public health threat. Foodborne illnesses are typically caused by bacteria, such as Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus, which are frequently resistant to common antimicrobial agents. Rice is a staple grain in most parts of the world. Our previous work showed that Phatthalung Sangyod rice seed protein hydrolysates (SYPs), especially SYP4, exhibit antifungal activity against several fungal species that are pathogenic for both humans and animals and are non-cytotoxic to animal red blood cells. In this study, we aimed to determine the effects of the bioactive peptides in SYPs against several pathogenic bacteria in humans and animals.
Materials and Methods
After isolating SYP1, it was treated as follows: heated (SYP2), and hydrolyzed using pepsin (SYP3), and proteinase K (SYP4). Then, we used 500 μg of protein to evaluate the antibacterial effects on four pathogenic bacteria, including E. coli, P. aeruginosa, B. cereus, and S. aureus, using agar well diffusion. Using a broth microdilution assay, we determined the minimum inhibitory and bactericidal concentration (MIC and MBC, respectively) values of active SYPs. Using the agar well diffusion and microtube incubation methods, we also assessed the inhibitory effects of SYPs on the bacterial quorum sensing (QS) activity of Chromobacterium violaceum. Sangyod rice seed protein hydrolysates were evaluated for their ability to inhibit the biofilm formation of bacterial cells by a crytal violet assay. Furthermore, using the dropping method, we tested the inhibitory effects of SYPs on the bacterial pigments pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus.
Results
Our results showed that the crude protein lysate (SYP1) did not exhibit antibacterial activity against any of the test bacteria. Intriguingly, after boiling (SYP2) and enzymatic hydrolysis (SYP3 and SYP4), the protein hydrolysates were transformed into bioactive peptides and displayed antibacterial properties against all of the test bacteria at a concentration of 500 μg as determined by agar well diffusion. SYP4 demonstrated the highest antibacterial activity as it completely inhibited all test strains, with inhibition zones ranging from 16.88 ± 0.25 to 21.25 ± 0.5 mm, and also yielded the highest MIC/MBC values against P. aeruginosa, B. cereus, and E. coli, at 256 and >256 μg/mL, respectively. We observed that at least 256 μg/mL of SYP4 is required to exhibit optimal antibacterial activity. At 16–128 μg/mL, it exhibited antibiofilm activity against S. aureus. Furthermore, at 256 μg/mL, SYP4 inhibited pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus. Although SYP2 and SYP3 displayed weak antibacterial activity and their MIC values could not be obtained for all bacteria, they showed strong QS inhibition in C. violaceum at 256 μg protein. Moreover, SYP2 and SYP3, at a minimum concentration of 32 μg/mL, significantly reduced violacein production. SYP3 also showed biofilm reduction activity on S. aureus at least 16–512 μg/mL.
Conclusion
Sangyod Phatthalung protein hydrolysates exerted excellent inhibitory effects against the growth of bacteria and their virulence factors, such as QS, biofilm formation, and/or pigment production. These factors include zoonotic and foodborne pathogens. Therefore, daily consumption of Sangyod Phatthalung rice might reduce the risk of bacterial pathogenesis and foodborne diseases. In conclusion, functional foods or alternate methods of treating bacterial illnesses may be developed in humans and animals.
... Brucea javanica (L.) Merr. is a traditional Chinese medicinal herb that has been used to treat dysentery, malaria, cancer, and other diseases in China and Southeast Asian coun- [72,73]. Yadanzi oil is extracted from the mature fruits of Brucea javanica and possesses multiple biological activities, serving as the main source of functional activity for Brucea javanica [74]. ...
Inflammatory bowel disease (IBD) is a chronic intestinal inflammatory disorder that includes ulcerative colitis (UC) and Crohn's disease (CD), the exact cause of which is still unknown. Numerous studies have confirmed that diet is one of the major environmental factors associated with IBD, as it can regulate the gut microbiota and reduce inflammation and oxidative stress. Since the consumption of oil is essential in the diet, improving IBD through oil has potential. In this article, we first briefly reviewed the current treatment methods for IBD and introduce the role of natural oils in improving inflammatory diseases. We then focused on the recent discovery of the role of natural oils in the prevention and treatment of IBD and summarized their main mechanisms of action. The results showed that the anti-inflammatory activity of oils derived from different plants and animals has been validated in various experimental animal models. These oils are capable of improving the intestinal homeostasis in IBD animal models through multiple mechanisms, including modulation of the gut microbiota, protection of the intestinal barrier, reduction in colonic inflammation, improvement in oxidative stress levels in the intestine, and regulation of immune homeostasis. Therefore, dietary or topical use of natural oils may have potential therapeutic effects on IBD. However, currently, only a few clinical trials support the aforementioned conclusions. This review emphasized the positive effects of natural oils on IBD and encouraged more clinical trials to provide more reliable evidence on the improvement of human IBD by natural oils as functional substances.
... Pepsin and proteinase K were used for proteolytic digestion. The proteolytic digestion assays were performed as previously described with minor modifications [24,26]. Briefly, the proteins were incubated with either proteinase K (Invitrogen, Thermo Fisher Scientific™, California, USA) or porcine pepsin (Sigma-Aldrich, Missouri, USA) at an enzyme-toprotein ratio of 1:25. ...
Background and Aim: Fungal zoonoses are an economic and public health concern because they can cause various degrees of morbidity and mortality in animals and humans. To combat this issue, alternative natural antifungals, such as products derived from rice protein hydrolysates or rice antifungal protein/peptide are being considered because they are highly bioactive and exhibit various functional properties. Thailand is a leading rice producer and exporter. Among the various cultivated rice varieties, Sangyod rice (Oryza sativa L.) is exclusively indigenous to Thailand’s Phatthalung province; it has a Thai geographical indication tag. Here, we investigated whether the Phatthalung Sangyod rice seeds have bioactive antifungal peptides.
Materials and Methods: Antifungal activity in four Sangyod rice seed extracts (SYPs) – namely, (1) the crude lysate, SYP1; (2) the heat-treated lysate, SYP2; (3) the heat- and pepsin digested lysate, SYP3; and (4) the heat- and proteinase K-digested lysate, SYP4 – was analyzed. Protein concentrations in these SYPs were determined using the Bradford assay. The total phenolic compound content was determined using the modified Folin–Ciocalteu method in a 96-well microplate. Then, the SYP protein pattern was determined using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Subsequently, using the agar well diffusion method, the antifungal properties of these SYPs were tested against ten medically important pathogenic fungi. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration values were determined for the active SYPs – SYP2-4. Finally, the clinical safety of SYP4 was determined using a hemolytic assay (using canine red blood cells [RBCs]).
Results: The crude lysate SYP1 did not show antifungal activity against any of the ten tested pathogenic fungi. Surprisingly, hydrolysates SYP2, SYP3, and SYP4 displayed antifungal properties against the ten tested pathogenic fungi. Thus, heat and enzymatic hydrolysis seem to transform the bioactivity of the crude protein extract – SYP1. Further, SYP4 shows the most effective antifungal activity. It completely inhibited Cryptococcus neoformans, Talaromyces marneffei yeast phase, Trichophyton mentagrophytes, and Trichophyton rubrum. A partial inhibitory action on Candida albicans and Microsporum gypseum was possessed while showing the least activity to C. neoformans. SYP4 was nontoxic to canine RBCs. Hemolysis of canine RBCs was undetectable at 1 × MIC and 2 × MIC concentrations; therefore, it can be safely used in further applications.
Conclusion: These results indicate that heat and proteinase K hydrolyzed SYP is a very potent antifungal preparation against animal and human fungal pathogens and it can be used in future pharmaceuticals and functional foods.
... Out of 50 compounds, fumarostelline and brucine had the highest docking scores, both at −8.5 kcal/mol (L − 1 & L − 7). In earlier research [53,54], the alkaloid brucine showed antibacterial and antidengue action. Compared to other licensed antivirals (remdesivir, ritonavir, lopinavir, oseltamivir, and ribavirin), molecules with higher binding energies (remdesivir −7.8 kcal/mol) may be better SARS-CoV-2 inhibitors (Table 2) [55]. ...
Despite treatments and vaccinations, it remains difcult to develop naturally occurring COVID-19 inhibitors. Here, our main objective is to fnd potential lead compounds from the retrieved alkaloids with antiviral and other biological properties that selectively target the main SARS-CoV-2 protease (M pro), which is required for viral replication. In this work, 252 alkaloids were aligned using Lipinski's rule of fve and their antiviral activity was then assessed. Te prediction of activity spectrum of substances (PASS) data was used to confrm the antiviral activities of 112 alkaloids. Finally, 50 alkaloids were docked with M pro. Furthermore, assessments of molecular electrostatic potential surface (MEPS), density functional theory (DFT), and absorption, distribution, metabolism, excretion, and toxicity (ADMET) were performed, and a few of them appeared to have potential as candidates for oral administration. Molecular dynamics simulations (MDS) with a time step of up to 100 ns were used to confrm that the three docked complexes were more stable. It was found that the most prevalent and active binding sites that limit M pro 'sactivity are PHE294, ARG298, and GLN110. All retrieved data were compared to conventional antivirals, fumarostelline, strychnidin-10-one (L-1), 2,3-dimethoxy-brucin (L-7), and alkaloid ND-305B (L-16) and were proposed as enhanced SARS-CoV-2 inhibitors. Finally, with additional clinical or necessary study, it may be able to use these indicated natural alkaloids or their analogs as potential therapeutic candidates.
... Out of 50 compounds, fumarostelline and brucine had the highest docking scores, both at −8.5 kcal/mol (L − 1 & L − 7). In earlier research [53,54], the alkaloid brucine showed antibacterial and antidengue action. Compared to other licensed antivirals (remdesivir, ritonavir, lopinavir, oseltamivir, and ribavirin), molecules with higher binding energies (remdesivir −7.8 kcal/mol) may be better SARS-CoV-2 inhibitors (Table 2) [55]. ...
Despite treatments and vaccinations, it remains difficult to develop naturally occurring COVID-19 inhibitors. Here, our main objective is to find potential lead compounds from the retrieved alkaloids with antiviral and other biological properties that selectively target the main SARS-CoV-2 protease (Mpro), which is required for viral replication. In this work, 252 alkaloids were aligned using Lipinski’s rule of five and their antiviral activity was then assessed. The prediction of activity spectrum of substances (PASS) data was used to confirm the antiviral activities of 112 alkaloids. Finally, 50 alkaloids were docked with Mpro. Furthermore, assessments of molecular electrostatic potential surface (MEPS), density functional theory (DFT), and absorption, distribution, metabolism, excretion, and toxicity (ADMET) were performed, and a few of them appeared to have potential as candidates for oral administration. Molecular dynamics simulations (MDS) with a time step of up to 100 ns were used to confirm that the three docked complexes were more stable. It was found that the most prevalent and active binding sites that limit Mpro’sactivity are PHE294, ARG298, and GLN110. All retrieved data were compared to conventional antivirals, fumarostelline, strychnidin-10-one (L-1), 2,3-dimethoxy-brucin (L-7), and alkaloid ND-305B (L-16) and were proposed as enhanced SARS-CoV-2 inhibitors. Finally, with additional clinical or necessary study, it may be able to use these indicated natural alkaloids or their analogs as potential therapeutic candidates.
... However, this plant is used as a medicine to prevent dysentery, diarrhea, and malaria. As stated in [27], the potions of its fruits produced a new antibacterial compound for Streptococcus pyogenes bacteria where the effective compound is the bitter-tasting alkaloid called brucine. This reference can be utilized as reasoning for predicting this plant as a candidate for natural antibiotics in this study. ...
Jamu is the traditional Indonesian herbal medicine system that is considered to have many benefits such as serving as a cure for diseases or maintaining sound health. A Jamu medicine is generally made from a mixture of several herbs. Natural antibiotics can provide a way to handle the problem of antibiotic resistance. This research aims to discover the potential of herbal plants as natural antibiotic candidates based on a machine learning approach. Our input data consists of a list of herbal formulas with plants as their constituents. The target class corresponds to bacterial diseases that can be cured by herbal formulas. The best model has been observed by implementing the Random Forest (RF) algorithm. For 10-fold cross-validations, the maximum accuracy, recall, and precision are 91.10%, 91.10%, and 90.54% with standard deviations 1.05, 1.05, and 1.48, respectively, which imply that the model obtained is good and robust. This study has shown that 14 plants can be potentially used as natural antibiotic candidates. Furthermore, according to scientific journals, 10 of the 14 selected plants have direct or indirect antibacterial activity.
... B. javanica is distributed widely throughout the tropical and subtropical zones of China, including Guangdong, Guangxi, Yunnan, and Fujian provinces. Fructus Bruceae is commonly used as medicinal herb in clinical practice in China to treat dysentery and malaria (Sakaki et al., 1986;Sornwatana et al., 2013). In addition, Fructus Bruceae is also recorded in the Chinese Pharmacopoeia for treating many diseases, including intestinal inflammation, diarrhea, malaria, and different types of cancer (Yan et al., 2017). ...
Brucea javanica (Ya-dan-zi in Chinese) is a well-known Chinese herbal medicine, which is traditionally used in Chinese medicine for the treatment of intestinal inflammation, diarrhea, malaria, and cancer. The formulation of the oil (Brucea javanica oil) has been widely used to treat various types of cancer. It has also been found that B. javanica is rich in chemical constituents, including quassinoids, triterpenes, alkaloids and flavonoids. Pharmacological studies have revealed that chemical compounds derived from B. javanica exhibit multiple bioactivities, such as anti-cancer, anti-bacterial, anti-diabetic, and others. This review provides a comprehensive summary on the pharmacological properties of the main chemical constituents presented in B. javanica and their underlying molecular mechanisms. Moreover, the review will also provide scientific references for further research and development of B. javanica and its chemical constituents into novel pharmaceutical products for disease management.
... The presence of an important residue (R or K) at the end of a tryptic peptide boosts ionization and, consequently, also boosts the signal intensity of mass spectral peaks. 13 Antibacterial proteins and peptides are almost all cationic, and they kill bacteria by permeabilizing or weakening their membranes. 14 Molecular docking is a well-studied approach for identifying potent peptides without excessive effort and investment in research to investigate the activity as a result of binding affinity (kcal/mol). ...
Bioactive peptides bring revolutions in the medical field and become an attractive target for new antibiotics. The bioactive peptides can be derived from the hydrolysis of protein. This study aims to explore the antibacterial peptide from venom hydrolysate of the Spitting cobra (Naja sumatrana). The venom protein was isolated from venom using a gel filtration technique. The digestion of venom protein was performed using trypsin with and without alkylation and reduction. After that, fractionation was carried out using a strong cation exchange solid-phase extraction, followed by antibacterial activity testing. The active peptides present in the active fractions were identified using HRMS. The protein hydrolysis with pretreatment using alkylation and reduction gave almost two-fold higher than that of untreated hydrolysis. Both the venom protein and the hydrolysate showed antibacterial activity against Escherichia coli but there was no activity against Staphylococcus aureus. Antibacterial activity has been detected in numerous cation exchange fractions against Escherichia coli and Staphylococcus aureus. The four best bioactive peptides were identified with amino sequences of VYGGDSR, YTPTNK, TQFSDR, and TFQDSR. In addition, molecular docking analysis revealed that the peptides showed high-affinity scores, and the model best matched the DNA gyrase subunit B by salt bridge formation, whereas hydrogen bond interactions proved inhibition of bacterial replication as an antibacterial mechanism.
... They accomplished this through the hydrolysing actions of enzymes alcalase and flavourzyme on the proteins present in the seeds of the plant. A separate study by Sornwatana et al., 2013 used a similar approach to generate antibacterial peptides from the fruit protein of Fructus Bruceae by peptin hydrolysis. This method is often preferred over the process of employing microorganisms to perform this hydrolysing mechanism. ...
Background
Food spoilage is caused by the undesirable growth of spoilage microorganisms in food products. This spoilage can lead to the global loss and waste of food. It is estimated that 1.3 billion tons of edible food produced for human consumption is either lost or wasted each year. Different preservation techniques have been developed to prevent this spoilage; however, the problem still occurs. Plants have the ability to produce compounds to protect themselves from the harsh environment. This has led to the exploitation of plant antimicrobial peptides (AMPs) which have been naturally extracted to study their activity against plant, food, and human pathogens. The chemical synthesis of such peptides has also grown in popularity over the years.
Scope and approach
This review will focus on the different techniques that can be applied to generate peptide sequences, produce them through various chemical and biological methods, and predict their activity. These approaches are reviewed for their ability to develop synthetic peptides (based on plant AMPs) with greater antimicrobial activity and characteristics essential for their application as potential food preservatives. The development of these synthetic peptides with reduced toxicity to human cells, improved activity and stability can be a possible solution to the continuous fight against food spoilage and food waste.
Key findings and conclusions
Synthetic antimicrobial peptides are being developed and modified in such a way to encompass potent and safe characteristics. This knowledge can be exploited for the potential application of such peptides in foods as preservative agents. A deeper understanding of their structure, function and mechanisms of action can be used to integrate them into food for the reduction of food spoilage, and consequently, of food waste. Although their development and production through the methods reviewed can generate peptides with suitable characteristics for reducing food spoilage, the cost of synthesis can be a drawback to such methods. Nevertheless, as the technologies improve and develop over time, the development of these synthetic AMPs can be fully exploited for their potential role in the food sector.
... Similar to isolated natural peptides, peptides hydrolyzed from proteins vary from those derived from higher organisms. Purification of peptides from the hydrolysis of pepsin protein from Brucea javanica fruit produced brucine, an antibacterial peptide with the amino acid sequence HTLCMAGGATY, with activity against Streptococcus pyogenes 16 times higher than penicillin (Sornwatana et al., 2013). Antibacterial peptides hydrolyzed from plant proteins obtained from the hydrolysis of pepsin against RuBisCO, which was isolated from alfalfa, have three short peptides (i.e., three, five, and six amino acids) with antibacterial activity against both gram-positive and gramnegative bacteria (Kobbi et al., 2015). ...
... The supernatant was separated, and its absorbance was measured using a UV-Vis spectrophotometer at a wavelength of 280 nm. The level of hydrolysis was expressed as the ratio of the percentage of absorbance after and before enzymatic hydrolysis (Sornwatana et al., 2013). ...
... A high degree of hydrolysis is unnecessary in peptide identification. Sornwatana et al. (2013) succeeded in identifying active peptides from protein hydrolysate, with a 3.7% degree of hydrolysis. However, a large amount of unhydrolyzed protein present in hydrolysate could hinder peptide identification. ...
Antimicrobial peptide (AMP) peptide-based lead compound has become interesting target in developing new antibiotics. AMP is possibly generated through the digestion of protein. The protein of castor (Ricinus communis) seed is characterized as a ribosome-inactivating protein (RIP) that can be a source of AMP. The objectives of this research are to identify antibacterial peptides from Ricinus communis seed protein hydrolysate. The seed protein was isolated using sodium dodecyl sulfate and subsequently digested using trypsin. The hydrolysate was fractionated using a strong cation exchange chromatography system, and the resulting fractions were tested for antibacterial activity. The peptides present in the active fraction were identified using high-resolution mass spectrometry. As the result, the pH 4 and pH 5 fractions of the elution buffer indicated antibacterial activity, with the pH 4 fraction of the hydrolysate having high activity against both gram-negative (Escherichia coli) and gram-positive (Staphylococcus aureus) bacteria. Three peptides that have the sequences EESETVGQR, GQSTGTGQQER, and LDALEPDNR could be responsible for the activity of the pH 4 fraction. The antibacterial activity of these peptides, which is due to their ionic properties and secondary structure, supports the disruption of the bacterial cell membrane. It can be concluded that Ricinus communis seed protein hydrolysate contains peptides with sequence EESETVGQR, GQSTGTGQQER, and LDALEPDNR that potent to be used as AMP lead compounds
