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用于生理学研究和药物发现的南极细菌嗜盐栖假交替单胞菌TAC125的大规模生物膜培养

Large-scale biofilm cultivation of Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 for physiologic studies and drug discovery.

作者信息

Parrilli Ermenegilda, Ricciardelli Annarita, Casillo Angela, Sannino Filomena, Papa Rosanna, Tilotta Marco, Artini Marco, Selan Laura, Corsaro Maria Michela, Tutino Maria Luisa

机构信息

Department of Chemical Sciences, Federico II University, Complesso Universitario Monte Sant'Angelo, Via Cintia 4, 80126, Naples, Italy.

Department of Public Health and Infectious Diseases, Sapienza University, Piazzale Aldo Moro 5, 00185, Rome, Italy.

出版信息

Extremophiles. 2016 Mar;20(2):227-34. doi: 10.1007/s00792-016-0813-2. Epub 2016 Feb 5.

Abstract

Microbial biofilms are mainly studied due to detrimental effects on human health but they are also well established in industrial biotechnology for the production of chemicals. Moreover, biofilm can be considered as a source of novel drugs since the conditions prevailing within biofilm can allow the production of specific metabolites. Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 when grown in biofilm condition produces an anti-biofilm molecule able to inhibit the biofilm of the opportunistic pathogen Staphylococcus epidermidis. In this paper we set up a P. haloplanktis TAC125 biofilm cultivation methodology in automatic bioreactor. The biofilm cultivation was designated to obtain two goals: (1) the scale up of cell-free supernatant production in an amount necessary for the anti-biofilm molecule/s purification; (2) the recovery of P. haloplanktis TAC125 cells grown in biofilm for physiological studies. We set up a fluidized-bed reactor fermentation in which floating polystyrene supports were homogeneously mixed, exposing an optimal air-liquid interface to let bacterium biofilm formation. The proposed methodology allowed a large-scale production of anti-biofilm molecule and paved the way to study differences between P. haloplanktis TAC125 cells grown in biofilm and in planktonic conditions. In particular, the modifications occurring in the lipopolysaccharide of cells grown in biofilm were investigated.

摘要

微生物生物膜主要因其对人类健康的有害影响而受到研究,但它们在工业生物技术的化学品生产中也已得到广泛应用。此外,生物膜可被视为新型药物的来源,因为生物膜内的环境条件能够促使特定代谢产物的产生。南极细菌嗜盐栖假交替单胞菌TAC125在生物膜条件下生长时会产生一种抗生物膜分子,能够抑制机会致病菌表皮葡萄球菌的生物膜形成。在本文中,我们建立了一种在自动生物反应器中培养嗜盐栖假交替单胞菌TAC125生物膜的方法。该生物膜培养旨在实现两个目标:(1)扩大无细胞上清液的产量,以满足抗生物膜分子纯化所需的量;(2)回收在生物膜中生长的嗜盐栖假交替单胞菌TAC125细胞用于生理学研究。我们建立了一种流化床反应器发酵方法,其中漂浮的聚苯乙烯载体均匀混合,形成最佳气液界面以促进细菌生物膜的形成。所提出的方法实现了抗生物膜分子的大规模生产,并为研究嗜盐栖假交替单胞菌TAC125在生物膜中生长与在浮游条件下生长的细胞之间的差异铺平了道路。特别是,研究了在生物膜中生长的细胞的脂多糖发生的变化。

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