Gut microbiota influence the host immune system and are associated with various diseases. In recent years, postmenopausal bone loss has been suggested to be related to gut microbiota. In the present study, we investigated the treatment effect of the probiotic Bacillus subtilis C-3102 (C-3102) on bone mineral density (BMD) and its influence on gut microbiota in healthy postmenopausal Japanese women. Seventy-six healthy postmenopausal Japanese women were treated with a placebo or C-3102 spore-containing tablets for 24 weeks. When compared with the placebo, C-3102 significantly increased total hip BMD (placebo = 0.83 ± 0.63%, C-3102 = 2.53 ± 0.52%, p=0.043). There was a significant group-by-time interaction effect for urinary type I collagen cross-linked N-telopeptide (uNTx) (p=0.033), a marker of bone resorption. Specifically, the C-3102 group showed significantly lower uNTx when compared with the placebo group at 12 weeks of treatment (p=0.015). In addition, in the C-3102 group, there was a trend towards a decrease in the bone resorption marker tartrate-resistant acid phosphatase isoform 5b (TRACP-5b) when compared with the placebo group at 12 weeks of treatment (p=0.052). The relative abundance of genus Bifidobacterium significantly increased at 12 weeks of treatment compared with the baseline in the C-3102 group. The relative abundance of genus Fusobacterium was significantly decreased in the C-3102 group at 12 and 24 weeks of treatment compared with the baseline. These data suggested that C-3102 improves BMD by inhibiting bone resorption and modulating gut microbiota in healthy postmenopausal women.
In the genome of Aspergillus oryzae, 12 genes have been predicted to encode serine-type carboxypeptidases. However, the carboxypeptidase activities of the proteins encoded by these genes have not yet been confirmed experimentally. In this study, we have constructed three of these 12 genes overexpressing strains using Aspergillus nidulans and characterized their overproduced recombinant proteins. Of these three genes, one was previously named cpI; the other two have not been reported yet, and hence, we named them ocpA and ocpB. The recombinant proteins released amino acid residues from the C terminus of peptides, and the activity of the enzymes was inhibited by phenylmethylsulfonyl fluoride, indicating the enzymes to be serine-type carboxypeptidases. Recombinant OcpA, OcpB, and CpI were stable at 45°C, 55°C, and 55°C, respectively, at a low pH. The enzymatic properties of recombinant OcpB were different from those of any reported serine-type carboxypeptidase. On the other hand, recombinant OcpA had similar enzymatic properties to A. oryzae carboxypeptidases O1 and O2. The DNA and N-terminal amino acid sequences of carboxypeptidases O1 and O2 from A. oryzae IAM2640 were similar to those of OcpA. Result of transcriptional analysis of ocpA, ocpB, and cpI suggest differences in transcriptional regulation between these genes.
ObjectiveTo investigate factors related to bifidobacterial colonization in early infancy, with a focus on maternal antimicrobial use at delivery.Study designA cross-sectional pilot study was performed. Feces samples of 33 Japanese healthy infants were collected over 10 months and analyzed by next-generation sequencing to examine the diversity and abundance of the gut microbiota.ResultsThe beta diversity index of the gut microbiota differed significantly based on maternal antimicrobial use at delivery (P < 0.05). The most dominant genus was bifidobacteria, and the relative abundance of bifidobacteria in infants exposed to maternal antibiotics was significantly lower than in those who were not exposed (P < 0.05). In contrast, the delivery mode showed no significant relationship with gut microbiota diversity.ConclusionsMaternal antimicrobial use at delivery has a stronger effect than delivery mode on the gut microbiota, especially for colonization of bifidobacteria.
Ingestion of Bacillus subtilis C-3102 spores (C-3102) has relieved the symptoms of diarrhoea in piglets and changed the composition of gut microbiota in humans. Recently, it was suggested that the composition of the human gut microbiota affects stool consistency. In this study, a double-blind, randomised, placebo-controlled trial was conducted to assess the preventive effects of chronic diarrhoea in healthy volunteers with loose stools by ingestion of C-3102. The results showed that oral doses of C-3102 tablets significantly decreased the Bristol Stool Scale score and stool frequency, and also significantly improved abdominal sounds. With regard to gut microbiota, the relative abundance of Lachnospira, Actinomyces and SMB53 were significantly changed. This study shows that C-3102 could be effective for treating loose stools (Trial registration: UMIN000022583, http://tinyurl.com/ya4refqn ).
Bifidobacterium species are known to fulfill important functions within the human colon. Thus, stimulating the activity of bifidobacteria is important to maintain host health. We revealed that culture supernatants of Bacillus subtilis C-3102 (referred to as C-3102) stimulated the growth of Bifidobacterium species. In this study, we isolated and identified six bifidogenic growth factors, which were cyclo (D-Val-D-Ile), cyclo (L-Val-D-Ile), cyclo (D-Val-L-Ile), cyclo (L-Val-L-Ile), cyclo (D-Val-L-Leu) and cyclo (L-Val-L-Leu). These six cyclic dipeptides increased the growth of Bifidobacterium species and had no effect on potentially harmful gut organisms. Moreover, supplementation with a mixture of these six cyclic dipeptides significantly increased the abundance of microorganisms related to the genus Bifidobacterium in a human colonic microbiota model culture system, although supplementation with a single type of dipeptide had no effect. These results show that cyclic dipeptides containing Val-Leu and Val-Ile produced by C-3102 could serve as bifidogenic growth factors in the gut microbial community. In the past few decades, the human gut microbiota has been reported to play an important role in the promotion of health, and several illnesses are known to be influenced by imbalances of gut microbiota 1. Therefore, manipulation of the gut microbiota is a clinical target for the treatment of gut microbiota-related diseases 2. Bifidobacterial populations belonging to the phylum Actinobacteria are the most dominant microbial group present in the healthy infant gut 3. The population of Bifidobacterium in the human gut decreases gradually from infancy to childhood, then remains relatively stable during an adult's life, and ultimately decreases in old age 4. Numerous health-promoting effects have been ascribed to certain strains of the genus Bifidobacterium, which are widely used as probiotics 5. For instance, one Japanese study reported that allergic children had less bifidobacteria compared to non-allergic children at an early stage, 4 months of age, and prenatal supplementation of bifidobacteria to mothers and postnatal supplementation to infants reduced the risk of allergies 6. It has also been reported that ingestion of Bifidobacterium breve has an anti-obesity effect in mice fed a high-fat diet 7 , and ingestion of Bifidobacterium infantis was effective in relieving symptom of irritable bowel syndrome in women 8,9. These findings suggest an important role for bifiodobacteria in regulating intestinal homeostasis. Some Bacillus subtilis strains are safe, survive passage through the human gastrointestinal tract, and do not induce any undesirable physiological effects in human subjects 10,11. One reported probiotic effect of B. subtilis was associated with inhibiting the growth of pathogenic bacteria such as Salmonella enterica to improve the growth of beneficial bacteria such as lactobacilli 12. In our previous study, B. subtilis C-3102 (hereinafter, referred to as C-3102) was tested using the TNO Gastro-Intestinal Mod...
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