Bacteriophages of Bacillus subtilis (natto) and Their Contamination in Natto Factories

Nagai, Tatsuo

doi:10.5772/33555

Cited by 8 publications

(6 citation statements)

References 24 publications

(22 reference statements)

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“…γ-PGA-degrading enzymes represent a serious industrial problem in modern Natto / Cheonggukjang production plants, since they compromise the typical viscous texture associated with the extremely long polymer chains. In those settings, degradation of high molecular weight γ-PGA is occasionally observed and related to phage infections [ 29 ]. In 2003 Kimura and collaborators isolated phage ΦNIT1 from spoiled Natto and used it to infect B .…”

Section: Introductionmentioning

confidence: 99%

γ-PGA Hydrolases of Phage Origin in Bacillus subtilis and Other Microbial Genomes

et al. 2015

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Poly-γ-glutamate (γ-PGA) is an industrially interesting polymer secreted mainly by members of the class Bacilli which forms a shield able to protect bacteria from phagocytosis and phages. Few enzymes are known to degrade γ-PGA; among them is a phage-encoded γ-PGA hydrolase, PghP. The supposed role of PghP in phages is to ensure access to the surface of bacterial cells by dismantling the γ-PGA barrier. We identified four unannotated B. subtilis genes through similarity of their encoded products to PghP; in fact these genes reside in prophage elements of B. subtilis genome. The recombinant products of two of them demonstrate efficient polymer degradation, confirming that sequence similarity reflects functional homology. Genes encoding similar γ-PGA hydrolases were identified in phages specific for the order Bacillales and in numerous microbial genomes, not only belonging to that order. The distribution of the γ-PGA biosynthesis operon was also investigated with a bioinformatics approach; it was found that the list of organisms endowed with γ-PGA biosynthetic functions is larger than expected and includes several pathogenic species. Moreover in non-Bacillales bacteria the predicted γ-PGA hydrolase genes are preferentially found in species that do not have the genetic asset for polymer production. Our findings suggest that γ-PGA hydrolase genes might have spread across microbial genomes via horizontal exchanges rather than via phage infection. We hypothesize that, in natural habitats rich in γ-PGA supplied by producer organisms, the availability of hydrolases that release glutamate oligomers from γ-PGA might be a beneficial trait under positive selection.

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Section: Introductionmentioning

confidence: 99%

γ-PGA Hydrolases of Phage Origin in Bacillus subtilis and Other Microbial Genomes

et al. 2015

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“…It is an industrially important microorganism used for its probiotic activities in humans [ 2 ] and for commercial enzyme production [ 3 ]. In addition, B. subtilis is one of the major fermenting bacteria in soybean-based fermented foods such as Korean Cheonggukjang (a fast-fermented soybean product), Deonjang (fermented soybean paste), Gochujang (hot red pepper paste), and Japanese Natto [ 4 , 5 ]. These types of soybean-based fermented foods are an integral part of the diet of Asian peoples and the market values of these products are increasing day by day [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

“…In order to handle the bacteriophage contamination issue in lactic acid bacterial fermentation, a large number of bacteriophage were isolated and characterized, which led to the development of classification schemes and antiphage strategies (adapted factory design, use of phage-resistant starters, and air control) currently in operation [ 9 , 10 , 11 ]. On the other hand, some studies have been conducted in B. subtilis -infecting bacteriophage mostly in Japanese Natto [ 5 , 12 , 13 , 14 ], and their effects on the fermentation process are not well known.…”

Section: Introductionmentioning

confidence: 99%

Complete Nucleotide Sequence Analysis of a Novel Bacillus subtilis-Infecting Bacteriophage BSP10 and Its Effect on Poly-Gamma-Glutamic Acid Degradation

Ghosh

Lee

Kang

et al. 2018

Viruses

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While the harmful effects of lactic acid bacterial bacteriophages in the dairy industry are well-established, the importance of Bacillus subtilis-infecting bacteriophages on soybean fermentation is poorly-studied. In this study, we isolated a B. subtilis-infecting bacteriophage BSP10 from Meju (a brick of dried fermented soybean) and further characterized it. This Myoviridae family bacteriophage exhibited a narrow host range against B. subtilis strains (17/52, 32.7%). The genome of bacteriophage BSP10 is 153,767 bp long with 236 open reading frames and 5 tRNAs. Comparative genomics (using dot plot, progressiveMauve alignment, heat-plot, and BLASTN) and phylogenetic analysis strongly suggest its incorporation as a new species in the Nit1virus genus. Furthermore, bacteriophage BSP10 was efficient in the growth inhibition of B. subtilis ATCC 15245 in liquid culture and in Cheonggukjang (a soybean fermented food) fermentation. Artificial contamination of as low as 102 PFU/g of bacteriophage BSP10 during Cheonggukjang fermentation significantly reduced bacterial numbers by up to 112 fold in comparison to the control (no bacteriophage). Moreover, for the first time, we experimentally proved that B. subtilis-infecting bacteriophage greatly enhanced poly-γ-glutamic acid degradation during soybean fermentation, which is likely to negatively affect the functionalities of Cheonggukjang.

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“…Bacillus subtilis ( B. subtilis ) natto is usually used to produce salt-free fermented soy food. It has been demonstrated that B. subtilis natto is activated to strengthen beneficial intestinal flora, inhibit harmful intestinal bacteria, produce protein and starch degradation enzymes, and increase the production of B-complex vitamins ( Nagai, 2012 ; Bhat et al, 2013 ). B. subtilis natto has been widely used as a probiotic in animal feeds ( Giang et al, 2011 ).…”

Section: Introductionmentioning

confidence: 99%

“…B. coagulans has been found to stimulate productive performance and improve meat quality in chickens ( Zhou et al, 2010 ). Previous studies mostly examined the efficacy of single probiotic in animal feed on growth performance and meat quality ( Zhou et al, 2010 ; Giang et al, 2011 ; Nagai, 2012 ; Bhat et al, 2013 ). The combined efficacy of two or multiple probiotics, such as B. subtilis natto and B. coagulans , in improving animal growth performance and meat quality has been rarely studied.…”

Section: Introductionmentioning

confidence: 99%

Effect of <i>Bacillus subtilis</i> Natto on Meat Quality and Skatole Content in TOPIGS Pigs

Sheng

Zhou

et al. 2015

Asian Australas. J. Anim. Sci

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This study investigated the effect of Bacillus subtilis (B. subtilis) natto on meat quality and skatole in TOPIGS pigs. Sixty TOPIGS pigs were randomly assigned to 3 groups (including 5 pens per group, with 4 pigs in each pen) and fed with basic diet (control group), basic diet plus 0.1% B. subtilis natto (B group), and basic diet plus 0.1% B. subtilis natto plus 0.1% B. coagulans (BB group), respectively. All pigs were sacrificed at 100 kg. Growth performance, meat quality, serum parameters and oxidation status in the three groups were assessed and compared. Most parameters regarding growth performance and meat quality were not significantly different among the three groups. However, compared with the control group, meat pH24, fat and feces skatole and the content of Escherichia coli (E. Coli), Clostridium, NH3-N were significantly reduced in the B and BB groups, while serum total cholesterol, high density lipoprotein, the levels of liver P450, CYP2A6, and CYP2E1, total antioxidant capability (T-AOC) and glutathione peroxidase and Lactobacilli in feces were significantly increased in the B and BB groups. Further, the combined supplementation of B. subtilis natto and B. coagulans showed more significant effects on the parameters above compared with B. subtilis, and Clostridium, and NH3-N. Our results indicate that the supplementation of pig feed with B. subtilis natto significantly improves meat quality and flavor, while its combination with B. coagulans enhanced these effects.

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Bacteriophages of Bacillus subtilis (natto) and Their Contamination in Natto Factories

Cited by 8 publications

References 24 publications

γ-PGA Hydrolases of Phage Origin in Bacillus subtilis and Other Microbial Genomes

γ-PGA Hydrolases of Phage Origin in Bacillus subtilis and Other Microbial Genomes

Complete Nucleotide Sequence Analysis of a Novel Bacillus subtilis-Infecting Bacteriophage BSP10 and Its Effect on Poly-Gamma-Glutamic Acid Degradation

Effect of <i>Bacillus subtilis</i> Natto on Meat Quality and Skatole Content in TOPIGS Pigs

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