Ralstonia eutropha H16 Flagellation Changes According to Nutrient Supply and State of Poly(3-Hydroxybutyrate) Accumulation

Author:

Raberg Matthias1,Reinecke Frank1,Reichelt Rudolf2,Malkus Ursula2,König Simone3,Pötter Markus1,Fricke Wolfgang Florian45,Pohlmann Anne6,Voigt Birgit7,Hecker Michael7,Friedrich Bärbel6,Bowien Botho4,Steinbüchel Alexander1

Affiliation:

1. Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, D-48149 Münster, Germany

2. Institut für Medizinische Physik und Biophysik, Elektronenmikroskopie und Analytik, Universitätsklinikum Münster, Westfälische Wilhelms- Universität, D-48149 Münster, Germany

3. Institut für Integrierte Funktionelle Genomik (IFG), Zentrum für Interdisziplinäre Klinische Forschung (IZKF), Medizinische Fakultät der Universität Münster, D-48149 Münster, Germany

4. Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstraße 8, D-37077 Göttingen, Germany

5. Göttingen Genomics Laboratory, Georg-August-Universität Göttingen, Grisebachstraße 8, D-37077 Göttingen, Germany

6. Institut für Biologie/Mikrobiologie, Humboldt-Universität zu Berlin, Chausseestraße 117, D-10115 Berlin, Germany

7. Institut für Mikrobiologie, Ernst-Moritz-Arndt Universität, Friedrich Ludwig Jahn Straße 15, D-17489 Greifswald, Germany

Abstract

ABSTRACT Two-dimensional polyacrylamide gel electrophoresis (2D PAGE), in combination with matrix-assisted laser desorption ionization-time of flight analysis, and the recently revealed genome sequence of Ralstonia eutropha H16 were employed to detect and identify proteins that are differentially expressed during different phases of poly(3-hydroxybutyric acid) (PHB) metabolism. For this, a modified protein extraction protocol applicable to PHB-harboring cells was developed to enable 2D PAGE-based proteome analysis of such cells. Subsequently, samples from (i) the exponential growth phase, (ii) the stationary growth phase permissive for PHB biosynthesis, and (iii) a phase permissive for PHB mobilization were analyzed. Among several proteins exhibiting quantitative changes during the time course of a cultivation experiment, flagellin, which is the main protein of bacterial flagella, was identified. Initial investigations that report on changes of flagellation for R. eutropha were done, but 2D PAGE and electron microscopic examinations of cells revealed clear evidence that R. eutropha exhibited further significant changes in flagellation depending on the life cycle, nutritional supply, and, in particular, PHB metabolism. The results of our study suggest that R. eutropha is strongly flagellated in the exponential growth phase and loses a certain number of flagella in transition to the stationary phase. In the stationary phase under conditions permissive for PHB biosynthesis, flagellation of cells admittedly stagnated. However, under conditions permissive for intracellular PHB mobilization after a nitrogen source was added to cells that are carbon deprived but with full PHB accumulation, flagella are lost. This might be due to a degradation of flagella; at least, the cells stopped flagellin synthesis while normal degradation continued. In contrast, under nutrient limitation or the loss of phasins, cells retained their flagella.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3