Laboratory of Chemical Biotechnology, Department of Biochemical and Chemical Engineering, TU Dortmund University, Emil-Figge-Strasse 66, 44227, Dortmund, Germany.
Department of Solar Materials, Helmholtz-Centre for Environmental Research - UFZ, Permoserstrasse 15, 04318, Leipzig, Germany.
Appl Microbiol Biotechnol. 2016 Jan;100(1):347-60. doi: 10.1007/s00253-015-7006-2. Epub 2015 Oct 1.
Five mutants of Pseudomonas taiwanensis VLB120ΔCeGFP showed significant autoaggregation when growing on defined carbohydrates or gluconate, while they grew as suspended cells on complex medium and on organic acids like citrate and succinate. Surprisingly, the respective mutations affected very different genes, although all five strains exhibited the same behaviour of aggregate formation. To elucidate the mechanism of the aggregative behaviour, the microbial adhesion to hydrocarbons (MATH) assay and contact angle measurements were performed that pointed to an increased cell surface hydrophobicity. Moreover, investigations of the outer layer of the cell membrane revealed a reduced amount of O-specific polysaccharides in the lipopolysaccharide of the mutant cells. To determine the regulation of the aggregation, reverse transcription quantitative real-time PCR was performed and, irrespective of the mutation, the transcription of a gene encoding a putative phosphodiesterase, which is degrading the global second messenger cyclic diguanylate, was decreased or even deactivated in all mutants. In summary, it appears that the trophic autoaggregation was regulated via cyclic diguanylate and a link between the cellular cyclic diguanylate concentration and the lipopolysaccharide composition of P. taiwanensis VLB120ΔCeGFP is suggested.
5 株假单胞菌 VLB120ΔCeGFP 的突变体在生长于特定碳水化合物或葡萄糖酸盐时表现出明显的自聚集,而在复杂培养基和柠檬酸、琥珀酸等有机酸上则呈悬浮细胞生长。令人惊讶的是,尽管所有 5 株菌都表现出相同的聚集形成行为,但各自的突变影响了非常不同的基因。为了阐明聚集行为的机制,进行了微生物烃类吸附(MATH)测定和接触角测量,这些测定表明细胞表面疏水性增加。此外,对细胞膜外层的研究表明,突变细胞的脂多糖中 O-特异性多糖的量减少。为了确定聚集的调控,进行了反转录定量实时 PCR,无论突变如何,编码一种假定磷酸二酯酶的基因的转录在所有突变体中均减少甚至失活,该磷酸二酯酶降解全局第二信使环二鸟苷酸。总之,似乎营养自聚集是通过环二鸟苷酸调控的,并且建议假单胞菌 VLB120ΔCeGFP 的细胞中环二鸟苷酸浓度与脂多糖组成之间存在联系。