Binding activity of Natto (a fermented food) and Bacillus natto isolates to mutagenic-carcinogenic heterocyclic amines

2001 ◽  
Vol 47 (10) ◽  
pp. 935-942 ◽  
Author(s):  
Rajam Rajendran ◽  
Yoshiyuki Ohta

The fermented food, whole meal Natto, viscous polymeric material from Natto, Natto bean, cooked soya bean, and 28 bacterial isolates from Natto were studied for their binding capacity to foodborne mutagenic-carcinogenic heterocyclic amines. The mutagenic heterocyclic amines used were Trp-P-1 (3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indole); Trp-P-2 (3-amino-1-methyl-5H-pyrido(4,3-b)indole); Glu-P-1 (2-amino-6-methyldipyrido(1,2-a:3'2'-d)imidazole); PhIP (2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine); IQ (2-amino-3-methylimidazo(4,5-f)quinoline); MeIQ (2-amino-3,4-dimethylimidazo(4,5-f)quinoxaline); MeIQx (2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline); and MeAαC (2-amino-3-methyl-9H-pyrido(2,3)indole). The lyophilized Natto and other fractions of Natto exhibited high binding activity towards Trp-P-1, Trp-P-2, PhIP, and MeAαC, while Glu-P-1, IQ, and MeIQ were not effectively bound. The binding capacity of bacterial isolates (Bacillus natto) were isolate-mutagen dependent. Heat treated lyophilized cells, cell wall, and cytoplasmic contents of the bacterial isolate with the highest binding capacity were analyzed for their ability to bind different heterocyclic amines. The results indicate the importance of the cell wall in binding to heterocyclic amines, whereas the cytoplasmic contents were less effective. Heat-treated cells were not much different from that of viable cells in their binding. The impact of different factors, such as pH, incubation time, metal ions, different concentrations of sodium chloride and alcohol, various enzymes, and acetylation of mutagens on binding of Trp-P-1 and IQ, were discussed. The significance of the present results is also discussed from the viewpoint that Natto, a fermented food, is able to scavenge dietary mutagenic heterocyclic amines through binding.Key words: fermented food, mutagens, heterocyclic amines, Natto, binding.

1998 ◽  
Vol 44 (2) ◽  
pp. 109-115 ◽  
Author(s):  
Rajam Rajendran ◽  
Yoshiyuki Ohta

Miso, a widely used Japanese fermented food was analysed for its lactic acid bacterial count on bromocresol purple agar. The binding of eight different foodborne carcinogenic heterocyclic amines to 25 bacterial isolates from miso were investigated. The heterocyclic amines used were 3-amino-1,4-dimethyl[5H]pyrido(4,3-b)indole (Trp-P-1), 3-amino-1-methyl[5H]pyrido(4,3-b)indole (Trp-P-2), 2-amino-6-methyldipyrido(1,2-a:3'2'-d)imidazole (Glu-P-1), 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP), 2-amino-dimethylimidazo(4,5f)quinoline (IQ), 2-amino-3,4-dimethylimidazo(4,5-f) quinoline (MeIQ), 2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline (MeIQx), and 2-amino-3-methyl-9H-pyrido(2,3)indole (MeAαC). The lyophilized cells of all of the isolates exhibited high binding activity towards Trp-P-1, Trp-P-2, MeAαC, and PhIP, while Glu-P-1 and IQ were not effectively bound. Of the isolates tested, the strongest and weakest binders were identified as Pediococcus acidilactici 1 and 2, respectively. Lyophilized cell wall fractions, heat-treated cells, and the cytoplasmic contents of P. acidilactici 1 and 2 were analysed for their ability to bind to different mutagens. Pure cell wall and peptidoglycan showed greater binding activity than the bacterial cells. Cytoplasmic content also showed some binding, but it was much less effective. The impact of enzymes (amylase, protease, cellulase, chitinase, muraminase, and peptidase) and acetylation of Trp-P-1 and IQ on the binding action of bacteria and cell wall material were also analysed to understand the possible processes involved in the binding of lactic acid bacteria to carcinogenic heterocyclic amines.Key words: mutagen, heterocyclic amines, lactic acid bacteria, binding, miso.


1988 ◽  
Vol 43 (9-10) ◽  
pp. 656-664
Author(s):  
Frank Beise ◽  
Harald Labischinski ◽  
Hans Bradaczek

Abstract The binding capacity of penicillin G-sulfoxide towards the penicillin-binding proteins (PBP) of Staphylococcus aureus H was studied. The sulfoxide and its parent compound, penicillin G , differ only in two aspects, the sulfur-bound oxygen and an altered conformation of the five-membered thiazolidine-ring system. These minor alterations of the penicillin structure resulted in a drastical decrease of binding activity (about two orders of magnitude) of the sulfoxide derivative towards its target enzymes. Furthermore, the sulfoxide did not exhibit the selectivity of subinhibitory doses for PBP 3, as could be observed for penicillin G . The biological consequences of this behaviour were monitored via growth curves, uptake of cell wall label, and analysis of the cell wall. Binding studies revealed that comparable growth inhibition and impairment of cell wall label uptake were achieved by at least a 100-fold higher penicillin G-sulfoxide concentration, compared to its parent compound. In cell wall analysis, the application of high doses of the antibiotics, i.e. nearly saturated PBP , verified the above mentioned observation. Surprisingly, small but significant differences in cell wall composition occurred using subinhibitory doses, probably due to the altered affinity towards PBP 3, supporting the hypothesis of an important role of this PBP in peptidoglycan transpeptidation.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Haruka Sugiura ◽  
Ayumi Nagase ◽  
Sayoko Oiki ◽  
Bunzo Mikami ◽  
Daisuke Watanabe ◽  
...  

Abstract Saprophytic bacteria and plants compete for limited nutrient sources. Bacillus subtilis grows well on steamed soybeans Glycine max to produce the fermented food, natto. Here we focus on bacterial responses in conflict between B. subtilis and G. max. B. subtilis cells maintained high growth rates specifically on non-germinating, dead soybean seeds. On the other hand, viable soybean seeds with germinating capability attenuated the initial growth of B. subtilis. Thus, B. subtilis cells may trigger saprophytic growth in response to the physiological status of G. max. Scanning electron microscope observation indicated that B. subtilis cells on steamed soybeans undergo morphological changes to form apertures, demonstrating cell remodeling during saprophytic growth. Further, transcriptomic analysis of B. subtilis revealed upregulation of the gene cluster, yesOPQR, in colonies growing on steamed soybeans. Recombinant YesO protein, a putative, solute-binding protein for the ATP-binding cassette transporter system, exhibited an affinity for pectin-derived oligosaccharide from plant cell wall. The crystal structure of YesO, in complex with the pectin oligosaccharide, was determined at 1.58 Å resolution. This study expands our knowledge of defensive and offensive strategies in interspecies competition, which may be promising targets for crop protection and fermented food production.


Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 245
Author(s):  
Hiroshi Sekiya ◽  
Maho Okada ◽  
Eiji Tamai ◽  
Toshi Shimamoto ◽  
Tadashi Shimamoto ◽  
...  

Clostridium perfringens is an often-harmful intestinal bacterium that causes various diseases ranging from food poisoning to life-threatening fulminant disease. Potential treatments include phage-derived endolysins, a promising family of alternative antimicrobial agents. We surveyed the genome of the C. perfringens st13 strain and identified an endolysin gene, psa, in the phage remnant region. Psa has an N-terminal catalytic domain that is homologous to the amidase_2 domain, and a C-terminal domain of unknown function. psa and gene derivatives encoding various Psa subdomains were cloned and expressed in Escherichia coli as N-terminal histidine-tagged proteins. Purified His-tagged full-length Psa protein (Psa-his) showed C. perfringens-specific lytic activity in turbidity reduction assays. In addition, we demonstrated that the uncharacterized C-terminal domain has cell wall-binding activity. Furthermore, cell wall-binding measurements showed that Psa binding was highly specific to C. perfringens. These results indicated that Psa is an amidase endolysin that specifically lyses C. perfringens; the enzyme’s specificity is highly dependent on the binding of the C-terminal domain. Moreover, Psa was shown to have a synergistic effect with another C. perfringens-specific endolysin, Psm, which is a muramidase that cleaves peptidoglycan at a site distinct from that targeted by Psa. The combination of Psa and Psm may be effective in the treatment and prevention of C. perfringens infections.


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Amandine Leroy ◽  
Xavier Falourd ◽  
Loïc Foucat ◽  
Valérie Méchin ◽  
Fabienne Guillon ◽  
...  

Abstract Background Biomass recalcitrance is governed by various molecular and structural factors but the interplay between these multiscale factors remains unclear. In this study, hot water pretreatment (HWP) was applied to maize stem internodes to highlight the impact of the ultrastructure of the polymers and their interactions on the accessibility and recalcitrance of the lignocellulosic biomass. The impact of HWP was analysed at different scales, from the polymer ultrastructure or water mobility to the cell wall organisation by combining complementary compositional, spectral and NMR analyses. Results HWP increased the kinetics and yield of saccharification. Chemical characterisation showed that HWP altered cell wall composition with a loss of hemicelluloses (up to 45% in the 40-min HWP) and of ferulic acid cross-linking associated with lignin enrichment. The lignin structure was also altered (up to 35% reduction in β–O–4 bonds), associated with slight depolymerisation/repolymerisation depending on the length of treatment. The increase in $${T}_{1\rho }^{H}$$ T 1 ρ H , $${T}_{HH}$$ T HH and specific surface area (SSA) showed that the cellulose environment was looser after pretreatment. These changes were linked to the increased accessibility of more constrained water to the cellulose in the 5–15 nm pore size range. Conclusion The loss of hemicelluloses and changes in polymer structural features caused by HWP led to reorganisation of the lignocellulose matrix. These modifications increased the SSA and redistributed the water thereby increasing the accessibility of cellulases and enhancing hydrolysis. Interestingly, lignin content did not have a negative impact on enzymatic hydrolysis but a higher lignin condensed state appeared to promote saccharification. The environment and organisation of lignin is thus more important than its concentration in explaining cellulose accessibility. Elucidating the interactions between polymers is the key to understanding LB recalcitrance and to identifying the best severity conditions to optimise HWP in sustainable biorefineries.


Author(s):  
Marios Kazasidis ◽  
Elisa Verna ◽  
Shuo Yin ◽  
Rocco Lupoi

AbstractThis study elucidates the performance of cold-sprayed tungsten carbide-nickel coating against solid particle impingement erosion using alumina (corundum) particles. After the coating fabrication, part of the specimens followed two different annealing heat treatment cycles with peak temperatures of 600 °C and 800 °C. The coatings were examined in terms of microstructure in the as-sprayed (AS) and the two heat-treated conditions (HT1, HT2). Subsequently, the erosion tests were carried out using design of experiments with two control factors and two replicate measurements in each case. The effect of the heat treatment on the mass loss of the coatings was investigated at the three levels (AS, HT1, HT2), as well as the impact angle of the erodents (30°, 60°, 90°). Finally, the response surface methodology (RSM) was applied to analyze and optimize the results, building the mathematical models that relate the significant variables and their interactions to the output response (mass loss) for each coating condition. The obtained results demonstrated that erosion minimization was achieved when the coating was heat treated at 600 °C and the angle was 90°.


2021 ◽  
Vol 31 (1) ◽  
Author(s):  
Farid Abd-El-Kareem ◽  
Ibrahim E. Elshahawy ◽  
Mahfouz M. M. Abd-Elgawad

Abstract Background Black root rot of strawberry plants caused by Rhizoctonia solani, Fusarium solani, and Pythium sp. is a serious disease in Egypt. Biocontrol agents have frequently proved to possess paramount and safe tools against many diseases. The impact of soil treatments with 3 Bacillus pumilus isolates on black root rot disease of strawberry plants caused by R. solani, F., and Pythium sp. under laboratory and field conditions was examined herein on the commonly used ‘Festival’ strawberry cultivar. To increase the bacterial adhesion and distribution on the roots, each seedling was dipped in bacterial cell suspension at 1 × 108 colony-forming units/ml of each separate bacterial isolate for 30 min then mixed with 5% Arabic gum. Results The tested B. pumilus isolates significantly reduced the growth area of these 3 fungi. The two bacterial isolates Nos. 2 and 3 reduced the growth area by more than 85.2, 83.6, and 89.0% for R. solani, F. solani, and Pythium sp., respectively. Likewise, the 3 bacterial isolates significantly (P ≤ 0.05) inhibited the disease under field conditions. Isolates Nos. 2 and 3 suppressed the disease incidence by 64.4 and 68.9% and disease severity by 65.3 and 67.3%, respectively. The fungicide Actamyl had effect similar to that of the 2 isolates. B. pumilus isolates significantly enhanced growth parameters and yields of strawberry plants; isolates Nos. 2 and 3 raised the yield by 66.7 and 73.3%, respectively. Conclusions Bacillus pumilus isolates could effectively manage the black rot disease in strawberry herein. Due to the significant impact of the root rot disease on strawberry yield, B. pumilus should be further tested to manage the disease on strawberry on large scale in Egypt.


Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1665
Author(s):  
Natalia Nikonorova ◽  
Evan Murphy ◽  
Cassio Flavio Fonseca de Lima ◽  
Shanshuo Zhu ◽  
Brigitte van de Cotte ◽  
...  

Auxin plays a dual role in growth regulation and, depending on the tissue and concentration of the hormone, it can either promote or inhibit division and expansion processes in plants. Recent studies have revealed that, beyond transcriptional reprogramming, alternative auxin-controlled mechanisms regulate root growth. Here, we explored the impact of different concentrations of the synthetic auxin NAA that establish growth-promoting and -repressing conditions on the root tip proteome and phosphoproteome, generating a unique resource. From the phosphoproteome data, we pinpointed (novel) growth regulators, such as the RALF34-THE1 module. Our results, together with previously published studies, suggest that auxin, H+-ATPases, cell wall modifications and cell wall sensing receptor-like kinases are tightly embedded in a pathway regulating cell elongation. Furthermore, our study assigned a novel role to MKK2 as a regulator of primary root growth and a (potential) regulator of auxin biosynthesis and signalling, and suggests the importance of the MKK2 Thr31 phosphorylation site for growth regulation in the Arabidopsis root tip.


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