Antimicrobial activity and comparative metabolomic analysis of Priestia megaterium strains derived from potato and dendrobium
Jiameng LiuYan-Tian LiangShanshan WangNuo JinJing SunCong LuYufeng SunShuying LiBei FanFengzhong Wang
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Abstract The growth of endophytic bacteria is influenced by the host plant and their secondary metabolites and activities. In this study, P-NA14 and D-HT207 were isolated from potato tuber and dendrobium stem respectively. They were both identified as Priestia megaterium . The antimicrobial activities and metabolomic of both strains were explored. For antimicrobial activities, results showed that strain P-NA14 exhibited a better inhibition effect on the pathogen of dendrobium, while strain D-HT207 exhibited a better inhibition effect on the pathogen of potato. The supernatant of strain P-NA14 showed an inhibition effect only on Staphylococcus aureus , while the sediment of strain D-HT207 showed an inhibition effect only on Escherichia coli . For metabolomic analysis, the content of L-phenylalanine in strain P-NA14 was higher than D-HT207, and several key downstream metabolites of L-phenylalanine were associated with inhibition of S_aureus including tyrosine, capsaicin, etc. Therefore, we speculated that the different antimicrobial activities between strain P-NA14 and strain D-HT207 were possibly related to the content of L-phenylalanine and its metabolites. This study provided a preliminary exploration of the fact why different activities appeared on the strains that were identified as the same species but isolated from different hosts at the metabolomic level.Keywords:
Bacillus megaterium
Strain (injury)
Using mixed sodium alginate and polyviny1 alcohol, immobilizing monosultap high effective degradation Bacillus megaterium, and designing orthogonal experiment to observeing the degradation ability to the soil of immobilized Bacillus megaterium, and comparing immobilized Bacillus megraterium and dissociative Bacillus megraterium in relative degradation active. After all the above experiment, an optimal immobilizing condition is drawn as below: Polyviny1 Alcohol concentration: 100/0; Sodium Alginate concentration: 1.50/0; CaCl2 concentration: 1.50/0; Bacillus megaterium concentration: 20/0.
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Degradation
Sodium alginate
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식품 및 발효공업의 균체 제거에 이용할 수 있는 새로운 미생물 응집체를 개발할 목적으로 토양에서 분리한 Bacillus megaterium 의 응집제 생산 특성을 조사한 결과 sucrose 2.0%, NaNO_3 0.3%, tryptone 0.01%, beef extract 0.01%, MgSO_4 0.05%, CaCO_3 0.005%가 최적 배양배지로 조사되었으며 최적배지에서의 응집활성은 257.8 unit로 기초배지에 비해 응집활성이 약 161% 증가하였다. 한편 본 균주는 탄소원 및 무기염의 첨가에 의해 응집활성이 크게 영향을 받았으나 질소원의 경우 다른 성분에 비해 영향이 크지 않은 것으로 조사되어 본 균주가 생산한 응집체는 균체외 다당류의 일종인 것으로 추측된다. 또한 조정제 응집제에 대한 정색반응 결과 생산 응집제를 조정제한 후 정색반응 실험 결과는 비환원성 당으로 fructose를 함유하지 않은 다당의 일종인 것으로 추측되었다.
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SUMMARY: A method for the preparation of protoplasts of Bacillus megaterium is described. The protoplasts obtained were very active metabolically and lysates of them were rich in polyribosomes. Methods for the detection of polyribosomes and of nascent peptides are detailed.
Bacillus megaterium
Protoplast
Polysome
Bacillus (shape)
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Protoplasts of two doubly auxotrophis strains of Bacillus megaterium were fused by nascent calcium phosphate or by polyethylene glycol treatment, and then allowed to revert to bacillary form on selection media. The colonies formed were analyzed and found to be mixed populations of individual bacteria apparently containing parental, recombinant, and segregating genotypes. The evidence suggested that segregation was transitory, lasting only for a few clonings and reisolations.
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Bacillaceae
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SUMMARY: 14C-Tetrachlorsalicylanilide (TCS) was taken up by Bacillus megaterium, becoming localized on the cytoplasmic membrane and was recoverable from the membrane by treatment with normal horse serum. Normal phase-dark B. megaterium protoplasts became phase-bright and distorted on addition of TCS; osmotically resistant ‘protoplasts’ in buffer were obtained from TCS-treated B. megaterium subsequently treated with lysozyme. Both these conditions were inhibited or annulled by serum.
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In this study, the ability of CYP109E1 from Bacillus megaterium DSM319 to metabolize cholesterol was investigated. This steroid was identified as a new substrate to be converted by CYP109E1 with adrenodoxin and adrenodoxin reductase as redox partners in vitro. The biotransformation was successfully reproduced in vivo by using Bacillus megaterium cells that overexpressed CYP109E1. To enhance the production of cholesterol derivatives, an Escherichia coli based whole-cell system that harbored CYP109E1 was established. This novel system showed a 3.3-fold higher activity than that of the B. megaterium system, yielding about 45 mg L-1 of these products. Finally, the reaction products were isolated and identified to be the highly important cholesterol derivatives 24(S)- and 25-hydroxycholesterol.
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Bacillus megaterium has been widely used as a research tool for decades. Its use is on the rise as a recombinant protein production host and as a bioremediation bacterium. Bacteriophages against this bacterium may have biotechnological applications. Here, we describe the novel podophage Palmer, which infects B. megaterium.
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Из почвы выделена бактерия, проявившая высокий уровень антимикробной активности, и установлена ее принадлежность к виду Bacillus megaterium. Дополнительно исследовано образование антибиотиков у девяти штаммов этого вида из коллекции ВКПМ. Установлено, что при глубинном культивировании в общей сложности у девяти из десяти различных штаммов B. megaterium образуются антибактериальные антибиотики, отличающиеся по спектру действия. Описаны физико-химические свойства пяти соединений, из которых три отнесены к группе антибиотиков-пептидов. Все пять соединений эффективны в отношении метициллин-резистентного штамма Staphylococcus aureus ИНА 00761, и показано, что три из них относятся к ранее неописанным соединениям. Кроме того, антибиотики из разных штаммов B. megaterium проявляют активность в отношении устойчивого к гликопептидным антибиотикам штамма Leuconostoc mesenteroides ВКПМ B-4177, а также к грамотрицательным бактериям Pseudomonas aeruginosa ATCC 27853 и Escherichia coli ATCC 25922.
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Bacillaceae
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Bacillus megaterium is a large spore-forming bacterium found widely in the environment. Phages infecting B. megaterium can be used as genetic tools to expand the array of uses of B. megaterium in research and industry. Here, we present the complete genome of Pavlov, a podophage infecting B. megaterium.
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