RESUMEN
We performed several experiments using three strains of Virgibacillus salexigens, namely, P2, NT N53, and C-20MoT (DSM 11483T), which were isolated from completely different sources, in relation to bacteriocin production ability. Results of whole-genome sequencing analysis revealed that all strains have very similar sequences encoding class IId bacteriocin. Although a partial amino acid sequence of the purified bacteriocin produced by strain P2 isolated from fermented food was previously reported, whole-genome sequencing and the N-terminal sequencing results in this study showed that its complete amino acid sequence consisted of 48 residues, which corresponded to that of the hypothetical bacteriocin encoded by the gene in Virgibacillus massiliensis strain Vm-5T (DSM 28587T) isolated from the human gut. From the results of 16S rRNA gene sequencing and whole-genome sequencing analyses, we taxonomically confirmed Vm-5T to be a strain of V. salexigens, and its broth culture showed antibacterial activity. Strain NT N53 isolated from the deep-sea floor produced two bacteriocins, namely, NTN-A and NTN-B. The results of N-terminal sequencing, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and whole-genome sequencing analyses showed that their amino acid sequences differed in only one residue, and NTN-A showed the same sequence as the bacteriocin produced by strain P2. Although strain C-20MoT isolated from a solar saltern had the coding sequence very similar to that of NTN-A, its broth culture showed no antibacterial activity. This finding suggests that class IId bacteriocin-producing or bacteriocin-gene-encoding V. salexigens strains are widely distributed in distinct environment sources with different geographical and material properties.
Asunto(s)
Bacteriocinas/genética , Virgibacillus/clasificación , Virgibacillus/genética , Secuencia de Aminoácidos , Antibacterianos/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Bacteriocinas/metabolismo , Microbiología Ambiental , Humanos , ARN Ribosómico 16S , Agua de Mar/microbiología , Análisis de Secuencia de ADN , Virgibacillus/metabolismo , Secuenciación Completa del GenomaRESUMEN
Background: Spirulina platensis contains several bioactive molecules such as phenol, flavonoid and phycocyanin pigments. This study unveils total phenol, flavonoid, antioxidant activity, phycocyanin content and evaluated encapsulation efficiency from Ocimum basilicum intervention on S. platensis. O. basilicum intervention aims to reduce unpleasant odors from S. platensis that will increase consumption and increase bioactive compounds. Methods: The intervention was carried out by soaking a S. platensis control sample (SP) in O. basilicum with a ratio of 1:4 (w/v) and it was then dried (DSB) and microencapsulated by freeze drying methods (MSB) using a combination of maltodextrin and gelatin. Total flavonoid and phenolic analysis with curve fitting analysis used a linear regression approach. Antioxidant activity of samples was analysed with the 2,2'-azino-bis-3-3thylbenzthiazoline-6-sulphonic acid (ABTS) method. Data were analysed using ANOVA at significance level (p < 0.05) followed by Tukey test models using SPSS v.22. Results: The result of this study indicated that O. basilicum intervention treatment (DSB) has the potential to increase bioactive compounds such as total phenol, antioxidant activity and phycocyanin, and flavonoid content. Intervention of O. basilicum on S. platensis (DSB) significantly increases total phenol by 49.5% and phycocyanin by 40.7%. This is due to the phenol and azulene compounds in O. basilicum which have a synergistic effect on phenol and phycocyanin in S. platensis. Microencapsulation using a maltodexrin and gelatin coating is effective in phycocyanin protection and antioxidant activity with an encapsulation efficiency value of 71.58% and 80.5%. Conclusion: The intervention of O. basilicum on S. platensis improved the total phenol and phycocyanin content and there is potential for a pharmaceutical product for a functional food and pharmaceutical product.
Asunto(s)
Ocimum basilicum , Ficocianina , Antioxidantes/farmacología , Flavonoides , Gelatina , Fenol , Fenoles , Polvos , SpirulinaRESUMEN
A natural antibacterial-substance-producing gram-positive bacterium was isolated from terasi shrimp paste, a popular fermented product in Indonesia. This strain, a spore-forming and strictly aerobic bacterium, was identified as Virgibacillus salexigens by 16S rRNA gene sequence analysis. The antibacterial substance purified from the precipitated product in the culture supernatant of the strain using ammonium sulfate showed a broad inhibition spectrum against gram-positive bacteria, including a typical foodborne bacterium, namely, Listeria monocytogenes. The antibacterial activity of the substance was inactivated by treatments with various proteolytic enzymes. It was stable after heating or pH treatment, and approximately 60% of the initial activity remained even after heating at 121 °C for 15 min. In addition, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis indicated that its monoisotopic mass weight was 5318.4 Da (M+H)(+). On the basis of the results obtained by the automated Edman degradation technique and MALDI-TOF MS analysis, the substance can be classified as a member of Class IId bacteriocins, but it could not be identified as any of the previously purified substances except for the putative bacteriocin predicted from the draft genome sequence data of gram-positive bacteria such as Virgibacillus and Bacillus strains.
Asunto(s)
Bacteriocinas/metabolismo , Bacteriocinas/farmacología , Microbiología de Alimentos , Bacterias Grampositivas/efectos de los fármacos , Virgibacillus/aislamiento & purificación , Virgibacillus/metabolismo , Aerobiosis , Bacteriocinas/aislamiento & purificación , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Concentración de Iones de Hidrógeno , Indonesia , Peso Molecular , Proteolisis , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Esporas Bacterianas/citología , Temperatura , Virgibacillus/clasificación , Virgibacillus/genéticaRESUMEN
BACKGROUND: The squid ink that is discarded as waste during processing can be effectively utilised as a gel enhancer in surimi gels, especially those prepared from dark-fleshed fish which have poor gel properties. It also acts as an antioxidant, inhibiting lipid oxidation. This investigation aimed to study the effect of melanin-free ink (MFI) from splendid squid (Loligo formosana) on properties and oxidative stability of surimi gel from sardine (Sardinella albella). RESULTS: MFI (0-0.1 g kg(-1) surimi) increased the breaking force and deformation of sardine surimi gel in a dose-dependent manner (P < 0.05). The addition of MFI had no effect on whiteness of surimi gels (P > 0.05). The expressible moisture content of gels decreased as the levels of MFI increased (P < 0.05). Based on a microstructure study, gel added with MFI at a level of 0.08 g kg(-1) surimi was denser and finer than that of the control (without MFI). Surimi gels with MFI had lower peroxide values, thiobarbituric acid reactive substances, nonanal and 2-decenal. CONCLUSION: MFI could improve the properties of sardine surimi gel. Additionally, it was able to prevent lipid oxidation in surimi gels during refrigerated storage.