Takeshi Zendo

Functional dairy products are attracting consumers’ attention, as they simultaneously have nutritional and health benefits. Hence, we aimed in this study to fortify a dairy product (yogurt) with phytosomes of extract from Hydnora abyssinica (a holoparasitic plant that has ethnobotanical value) and a potential bacteriocin-producing probiotic lactic acid bacterium (LAB). Goat cheese was screened for LAB with promising antimicrobial activity, and the safety and probiotic potential of the selected isolate were studied. As a result, strain GA5 was selected due to its wide antimicrobial activity that was suggested to be caused by bacteriocin production. Moreover, this strain showed promising stress tolerance, in vitro antioxidant activity (95 ± 2.8%), and hydrophobic potential (87.18 ± 3.43%). Strain GA5 was molecularly identified as Lactiplantibacillus plantarum GA5. On the other hand, a hydromethanolic extract was prepared from H. abyssinica flowers, and its prebiotic potential and polyphenol content were evaluated. This extract was also encapsulated in phytosomes. Then, the physical and morphological characteristics of prepared phytosomes were studied. Yogurt fortified with these ingredients (L. plantarum GA5 together with free H. abyssinica extract or its extract encapsulated in phytosomes) showed higher antioxidant content, viscosity, texture profile, and sensory properties than the control. Furthermore, the yogurt remained unspoiled for over 21 days, indicating that the added ingredients prolonged its shelf life. As far as we know, this is the first study describing the fortification of yogurt with H. abyssinica phytosomes and a potential bacteriocin-producing probiotic LAB.

Streptococcus mutans is a cariogenic bacterium that produces a variety of bacteriocins and retains resistance to these bacteriocins. In this study, we investigated the susceptibility of 127 S . mutans strains to nukacins produced by Staphylococcus spp., which are commensal bacteria in humans. We detected diverse susceptibilities among strains. Nineteen strains had a disrupted LctF (type I), which is responsible for nukacin susceptibility, whereas the remaining 108 strains had an intact LctF (type II) and displayed resistance to nukacins. However, the type I strains still showed resistance to nukacins to some extent. Interestingly, 18/19 (94.7%) type I strains carried a mukA-T locus, which is related to the synthesis of mutacin K8, and mukFEG , an ABC transporter. In contrast, among type II strains, only 6/108 strains (5.6%) had both the mukA-T locus and mukFEG , 19/108 strains (17.6%) carried only mukFEG , and 83/108 strains (76.9%) harbored neither mukA-T nor mukFEG . We also found that MukF had two variants: 305 amino acids (type α) and 302 amino acids (type β). All type I strains showed a type α (MukFα), whereas most type II strains with mukFEG (22/25 strains) had a type β (MukFβ). Then, we constructed a mukFEG -deletion mutant complemented with MukFαEG or MukFβEG and found that only MukFαEG was involved in nukacin resistance. The nukacin resistance capability of type II-LctFEG was stronger than that of MukFαEG. In conclusion, we identified a novel nukacin resistance factor, MukFEG, and either LctFEG or MukFEG was active in most strains via genetic polymorphisms depending on mukA-T genes. IMPORTANCE Streptococcus mutans is an important pathogenic bacterium not only for dental caries but also for systemic diseases. S. mutans is known to produce a variety of bacteriocins and to retain resistance these bacteriocins. In this study, two ABC transporters, LctFEG and MukFEG, were implicated in nukacin resistance and each ABC transporter has two subtypes, active and inactive. Of the two ABC transporters, only one ABC transporter was always resistant, while the other ABC transporter was inactivated by genetic mutation. Interestingly, this phenomenon was defined by the presence or absence of the mutacin K8 synthesis gene region, one of the bacteriocins of S. mutans . This suggests that the resistance acquisition is tightly controlled in each strain. This study provides important evidence that the insertion of bacteriocin synthesis genes is involved in the induction of genetic polymorphisms and suggests that bacteriocin synthesis genes may play an important role in bacterial evolution.