Literature DB >> 34179275

Determination of Polyhydroxybutyrate (PHB) Content in Ralstonia eutropha Using Gas Chromatography and Nile Red Staining.

Janina R Juengert1, Stephanie Bresan1, Dieter Jendrossek1.   

Abstract

Ralstonia eutropha H16 produces and mobilizes (re-utilizes) intracellular polyhydroxybutyrate (PHB) granules during growth. This protocol describes the visualization of intracellular Nile red stained PHB granules and the quantification of PHB by gas chromatography. Our first method describes how to analyze PHB granules by fluorescence microscopy qualitatively. Our second approach enables the conversion of PHB to volatile hydroxycarboxylic acid methyl esters by acidic methanolysis and their quantification by gas chromatography. Through this method, it is possible to obtain an absolute quantification of PHB, e.g., per cell dry weight.
Copyright © 2018 The Authors; exclusive licensee Bio-protocol LLC.

Entities:  

Keywords:  Acidic methanolysis; Gas chromatography; Nile red; Polyhydroxybutyrate (PHB); Ralstonia eutropha

Year:  2018        PMID： 34179275      PMCID： PMC8203877          DOI： 10.21769/BioProtoc.2748

Source DB:  PubMed          Journal:  Bio Protoc        ISSN： 2331-8325


  12 in total

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Authors:  Alessandra Zonari; Thaís M M Martins; Ana Cláudia C Paula; Jankerle N Boeloni; Silviene Novikoff; Alexandra P Marques; Vitor M Correlo; Rui L Reis; Alfredo M Goes
Journal:  Acta Biomater       Date:  2015-02-07       Impact factor: 8.947

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3.  Pseudomonas oleovorans as a Source of Poly(beta-Hydroxyalkanoates) for Potential Applications as Biodegradable Polyesters.

Authors:  H Brandl; R A Gross; R W Lenz; R C Fuller
Journal:  Appl Environ Microbiol       Date:  1988-08       Impact factor: 4.792

4.  Polyhydroxyalkanoates production with Ralstonia eutropha from low quality waste animal fats.

Authors:  Sebastian L Riedel; Stefan Jahns; Steven Koenig; Martina C E Bock; Christopher J Brigham; Johannes Bader; Ulf Stahl
Journal:  J Biotechnol       Date:  2015-09-30       Impact factor: 3.307

5.  Effect of enzymatic degradation of chitosan in polyhydroxybutyrate/chitosan/calcium phosphate composites on in vitro osteoblast response.

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Journal:  J Mater Sci Mater Med       Date:  2016-10-21       Impact factor: 3.896

Review 6.  Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates.

Authors:  A J Anderson; E A Dawes
Journal:  Microbiol Rev       Date:  1990-12

Review 7.  Medical application of microbial biopolyesters polyhydroxyalkanoates.

Authors:  Qiong Wu; Yang Wang; Guo-Qiang Chen
Journal:  Artif Cells Blood Substit Immobil Biotechnol       Date:  2009-01-08

8.  A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds.

Authors:  P Spiekermann; B H Rehm; R Kalscheuer; D Baumeister; A Steinbüchel
Journal:  Arch Microbiol       Date:  1999-01       Impact factor: 2.552

9.  A closer look on the polyhydroxybutyrate- (PHB-) negative phenotype of Ralstonia eutropha PHB-4.

Authors:  Matthias Raberg; Birgit Voigt; Michael Hecker; Alexander Steinbüchel
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10.  Polyhydroxyalkanoate (PHA) Granules Have no Phospholipids.

Authors:  Stephanie Bresan; Anna Sznajder; Waldemar Hauf; Karl Forchhammer; Daniel Pfeiffer; Dieter Jendrossek
Journal:  Sci Rep       Date:  2016-05-25       Impact factor: 4.379
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  6 in total

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Review 2.  Potential Use of Microbial Enzymes for the Conversion of Plastic Waste Into Value-Added Products: A Viable Solution.

Authors:  Muhammad Tamoor; Nadia A Samak; Yunpu Jia; Muhammad Umar Mushtaq; Hassan Sher; Maryam Bibi; Jianmin Xing
Journal:  Front Microbiol       Date:  2021-11-30       Impact factor: 5.640

3.  High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741T and in silico analyses of the genus specific PhaC2 polymerase variant.

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4.  Identification of New Halomonas Strains from Food-related Environments.

Authors:  Ayaka Tsuji; Yasuko Takei; Taku Nishimura; Yoshinao Azuma
Journal:  Microbes Environ       Date:  2022       Impact factor: 2.912

5.  Study on the production of high 3HV content PHBV via an open fermentation with waste silkworm excrement as the carbon source by the haloarchaeon Haloferax mediterranei.

Authors:  Shuangfeng Cai; Yaran Wu; Runjie Liu; Hongzhe Jia; Yunxiao Qiu; Min Jiang; Yuwen Ma; Xingxu Yang; Siyu Zhang; Yan Zhao; Lei Cai
Journal:  Front Microbiol       Date:  2022-08-17       Impact factor: 6.064

6.  Mameliella sediminis sp. nov., a novel polyhydroxyalkanoate-accumulating bacterium.

Authors:  Wei-Shuang Zheng; Sheng-Qiang Zhai; Meng-Jun Zhang; Yi Huang
Journal:  Int J Syst Evol Microbiol       Date:  2022-03       Impact factor: 2.689
  6 in total

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