scholarly journals Isolation, Identification and Antimicrobial Evaluation of Bactericides Secreting Bacillus subtilis Natto as a Biocontrol Agent

Processes ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 259 ◽  
Author(s):  
Jing Zhang ◽  
Muhammad Bilal ◽  
Shuai Liu ◽  
Jiaheng Zhang ◽  
Hedong Lu ◽  
...  

Herein, a bactericide-secreting Bacillus strain, potentially useful as a biocontrol agent, was isolated from the commercial Yanjing Natto food. Following the biochemical and physiological evaluation, the molecular identification was performed using 16S rDNA sequencing of polymerase chain reaction-amplified DNA that confirmed the natto isolate as Bacillus subtilis natto (B. subtilis natto). The biocontrol (microbial inhibitory) capability of B. subtilis natto was investigated against Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, and yeast (Yarrowia lipolytica) and recorded. The antimicrobial activity of B. subtilis natto was further enhanced by optimizing the growth medium for optimal bactericides secretion. Under optimized conditions, B. subtilis natto exhibited much higher inhibitory activity against S. aureus with a zone of inhibition diameter up to 27 mm. After 48 h incubation, the optimally yielded B. subtilis natto broth was used to extract and purify the responsible bactericides by silica gel column chromatography, gel column chromatography, and semi-preparative high-performance liquid chromatography. Structural identification of purified bactericides (designated as NT-5, NT-6, and NT-7) from B. subtilis natto was performed by 13C-nuclear magnetic resonance (NMR) and mass spectral analyses. The NMR comparison also revealed that NT-5, NT-6, and NT-7 had identical structures, except for the fatty chain. In summary, the present study suggests the improved biocontrol and/or microbial inhibitory potential of newly isolated bactericides secreting B. subtilis natto.

2023 ◽  
Vol 83 ◽  
Author(s):  
D. Pei ◽  
Q. Zhang ◽  
X. Zhu ◽  
S. Han

Abstract Endophytic bacteria serve key roles in the maintenance of plant health and growth. Few studies to date, however, have explored the antagonistic and plant growth-promoting (PGP) properties of Prunus cerasifera endophytes. To that end, we isolated endophytic bacteria from P. cerasifera tissue samples and used a dual culture plate assay to screen these microbes for antagonistic activity against Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum, and F. moniliforme. Of the 36 strains of isolated bacteria, four (strains P1, P10, P16, and P20) exhibited antagonistic effects against all five model pathogens, and the P10 strain exhibited the strongest antagonistic to five pathogens. This P10 strain was then characterized in-depth via phenotypic assessments, physiological analyses, and 16s rDNA sequencing, revealing it to be a strain of Bacillus subtilis. Application of a P10 cell suspension (1×108 CFU/mL) significantly enhanced the seed germination and seedling growth of tomato in a greenhouse setting. This P10 strain further significantly suppressed tomato Verticillium wilt with much lower disease incidence and disease index scores being observed following P10 treatment relative to untreated plants in pot-based experiments. Tomato plants that had been treated with strain P10 also enhanced defense-related enzymes, peroxidase, superoxide dismutase, and catalase activity upon V. dahliae challenge relative to plants that had not been treated with this endophytic bacterium. The results revealed that the P10 bacterial strain has potential value as a biocontrol agent for use in the prevention of tomato Verticillium wilt.


2016 ◽  
Vol 5 (01) ◽  
pp. 4701
Author(s):  
Shobha R. I. ◽  
Andallu B.

Nature has provided mankind with a broad and structurally diverse array of pharmacologically active chemical compounds, phytoceuticals, which have proved to be indispensable for the cure of chronic diseases or as lead structures for novel therapeutic agents. Almost, 70% of modern medicines in India have been developed from plants used in the traditional system of medicine. With the technological advances and the development of more sophisticated isolation and analytical techniques, there is great scope for further systematic research to screen and isolate many more phytoceuticals which might be more effective/as effective as synthetic drugs and thereby assess their potential in protecting against chronic diseases. The present investigation is aimed at isolation and identification of phytoceuticals in a spice, aniseed (Pimpinella anisum L), therapeutically less-exploited and widely used only for culinary purpose, using analytical techniques viz. column chromatography, high performance thin layer chromatography (HPTLC.) and nuclear magnetic resonance spectrometry. Among all the solvent fractions of methanolic extract tested, methanolic extract and ethyl acetate fraction possessed highest amounts of bioactive compounds viz. phenolics, flavonoids, flavonols which was confirmed by qualitative, quantitative and HPTLC. analyses. Spectral analysis using NMR. of one of the sub-fractions of aniseeds obtained by column chromatography, revealed the presence of a glycosylated flavone, luteolin-6C-glucoside in aniseeds.  The present study revealed aniseeds to be a great source of bioactive phytoceuticals which can be novel candidates for development of new therapeutic agents.


AMB Express ◽  
2013 ◽  
Vol 3 (1) ◽  
pp. 36 ◽  
Author(s):  
Aditya R Bhat ◽  
Victor U Irorere ◽  
Terry Bartlett ◽  
David Hill ◽  
Gopal Kedia ◽  
...  

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1519
Author(s):  
Meinan Chang ◽  
Fengtao Ma ◽  
Jingya Wei ◽  
Junhao Liu ◽  
Xuemei Nan ◽  
...  

Previous studies have shown that Bacillus subtilis natto affects rumen fermentation and rumen microbial community structure, which are limited to detect a few microbial abundances using traditional methods. However, the regulation of B. subtilis natto on rumen microorganisms and the mechanisms of microbiota that affect rumen fermentation is still unclear. This study explored the effects of live and autoclaved B. subtilis natto on ruminal microbial composition and diversity in vitro using 16S rRNA gene sequencing and the underlying mechanisms. Rumen fluid was collected, allocated to thirty-six bottles, and divided into three treatments: CTR, blank control group without B. subtilis natto; LBS, CTR with 109 cfu of live B. subtilis natto; and ABS, CTR with 109 cfu of autoclaved B. subtilis natto. The rumen fluid was collected after 0, 6, 12, and 24 h of fermentation, and pH, ammonia nitrogen (NH3-N), microbial protein (MCP), and volatile fatty acids (VFAs) were determined. The diversity and composition of rumen microbiota were assessed by 16S rRNA gene sequencing. The results revealed LBS affected the concentrations of NH3-N, MCP, and VFAs (p < 0.05), especially after 12 h, which might be attributed to changes in 18 genera. Whereas ABS only enhanced pH and NH3-N concentration compared with the CTR group (p < 0.05), which might be associated with changes in six genera. Supplementation with live B. subtilis natto improved ruminal NH3-N and propionate concentrations, indicating that live bacteria were better than autoclaved ones. This study advances our understanding of B. subtilis natto in promoting ruminal fermentation, providing a new perspective for the precise utilization of B. subtilis natto in dairy rations.


2017 ◽  
Vol 184 (3) ◽  
pp. 838-851 ◽  
Author(s):  
Gabrielly Terassi Bersaneti ◽  
Nicole Caldas Pan ◽  
Cristiani Baldo ◽  
Maria Antonia Pedrine Colabone Celligoi

1998 ◽  
Vol 45 (8) ◽  
pp. 494-498
Author(s):  
Kanako MURAMATSU ◽  
Noriko YAMAWAKE ◽  
Kan KIUCHI

Author(s):  
Anggia Murni ◽  
Novriyandi Hanif ◽  
Masaki Kita ◽  
Latifah K. Darusman

Objective: To isolate and elucidate a cytotoxic principle against breast tumor MCF-7 cells of the Indonesian terrestrial plant Ficus deltoidea Jack leaves.Methods: F. deltoidea leaves collected at National Park of mount Gede-Pangrango, Indonesia have been subjected to chemical and biological work. F. deltoidea leaves were extracted with 96% aqueous ethanol (EtOH) and was then partitioned into three layers n-hexane, dichloromethane (CH2Cl2), and n-butanol (n-BuOH). All layers were checked for their activity against breast tumor MCF-7 cells using MTT assay method. A portion of the most active layer was purified using open column chromatography to give fraction that has toxicity against zebra fish embryos. Based on the assay-guided isolation, compound 1 was isolated. The chemical structure of 1 was elucidated using nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS) data as well as comparing data with literature.Results: The CH2Cl2 layer of F. deltoidea leaves was found to inhibit breast tumor MCF-7 cells with IC50 10 µg/ml which was the most toxic among the layers. A portion of the most active layer was purified using open column chromatography to give 7 fractions. The fraction 5 showed toxicity against zebrafish embryos (LC50 35 µg/ml, 48 hpf). This fraction was purified using high performance liquid chromatography (HPLC) octadecylsilyl (ODS) column with gradient elution 70% aqueous acetonitrile (MeCN) to 100% MeCN (linear gradient) for 40 min with UV detection at 254 nm (tR = 30.99 min) to give compound 1. The chemical structure of 1 was revealed as a chlorin-type compound named methyl 10-epi-pheophorbide A.Conclusion: Methyl 10-epi-pheophorbide A was isolated for the first time from the active fraction of the Indonesian F. deltoidea leaves or tabat barito. The chemical structure including absolute stereo chemistry was elucidated using NMR and HRMS data as well as by comparison with the literature values. The 13C NMR data has been added to complete the previous report.


2011 ◽  
Vol 8 (1) ◽  
pp. 305-311 ◽  
Author(s):  
Priyanka Kamaria ◽  
N. Kawathekar ◽  
Prerna Chaturvedi

In order to develop new antimicrobial agents, a series of Schiff bases of indole-3-aldehyde were synthesized by microwave assisted synthesis by takingDMFas solvent and evaluated for their antimicrobial activity. All the synthesized compounds were characterized byIR,1HNMRand mass spectral analysis. All compounds were tested against five gram positive and five gram negative bacterial strains and one fungal strain. All compounds exhibited better activity against gram positive strains than against gram negative strains and the compounds were found more active againstS.aureusandB.subtilis.


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