Production of Short Side Chain-Poly[Hydroxyalkanoate] by a Newly Isolated Ralstonia Pickettii Strain

Bonatto, Diego; Matias, Fernanda; Lisbôa, Márcia Pagno; Bogdawa, Heike Marlis; Henriques, João Antônio Pêgas

doi:10.1023/b:wibi.0000033063.55133.e1

Cited by 19 publications

(5 citation statements)

References 40 publications

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“…Two labelled proteins involved in PHA synthesis were identified, namely acetoacetyl‐CoA reductase NAD(P)H consuming and phasin proteins, which are involved in PHA granule stabilization (Neumann et al ., ). The latter were assigned to Ralstonia , a common representative of PHA‐producing bacteria (Bonatto et al ., ). During our experiment, high toluene concentrations were present in the PFR, and the aerobic degraders might have formed PHA granules in deprivation of oxygen as electron acceptor.…”

Section: Discussionmentioning

confidence: 99%

In situ protein‐SIP highlights Burkholderiaceae as key players degrading toluene by para ring hydroxylation in a constructed wetland model

Lünsmann

Kappelmeyer

Benndorf

et al. 2016

Environmental Microbiology

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In constructed wetlands, organic pollutants are mainly degraded via microbial processes. Helophytes, plants that are commonly used in these systems, provide oxygen and root exudates to the rhizosphere, stimulating microbial degradation. While the treatment performance of constructed wetlands can be remarkable, a mechanistic understanding of microbial degradation processes in the rhizosphere is still limited. We investigated microbial toluene removal in a constructed wetland model system combining 16S rRNA gene sequencing, metaproteomics and (13) C-toluene in situ protein-based stable isotope probing (protein-SIP). The rhizospheric bacterial community was dominated by Burkholderiales and Rhizobiales, each contributing about 20% to total taxon abundance. Protein-SIP data revealed that the members of Burkholderiaceae, the proteins of which showed about 73% of (13) C-incorporation, were the main degraders of toluene in the planted system, while the members of Comamonadaceae were involved to a lesser extent in degradation (about 64% (13) C-incorporation). Among the Burkholderiaceae, one of the key players of toluene degradation could be assigned to Ralstonia pickettii. We observed that the main pathway of toluene degradation occurred via two subsequent monooxygenations of the aromatic ring. Our study provides a suitable approach to assess the key processes and microbes that are involved in the degradation of organic pollutants in complex rhizospheric ecosystems.

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Section: Discussionmentioning

confidence: 99%

In situ protein‐SIP highlights Burkholderiaceae as key players degrading toluene by para ring hydroxylation in a constructed wetland model

Lünsmann

Kappelmeyer

Benndorf

et al. 2016

Environmental Microbiology

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“…and archaea (e.g. Haloarchaea ), intracellularly produce PHA granules [ 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 ], as storage of carbon and energy under limiting conditions of essential nutrients such as nitrogen or phosphate and in the presence of an excess of carbon source [ 13 , 15 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

Improved fermentation strategies in a bioreactor for enhancing poly(3-hydroxybutyrate) (PHB) production by wild type Cupriavidus necator from fructose

Nygaard

Yashchuk

Noseda

et al. 2021

Heliyon

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Poly(3-hydroxybutyrate) (PHB) belongs to the family of polyhydroxyalkanoates, biopolymers used for agricultural, industrial, or even medical applications. However, scaling up the production is still an issue due to the myriad of parameters involved in the fermentation processes. The present work seeks, firstly, to scale up poly(3-hydroxybutyrate) (PHB) production by wild type C. necator ATCC 17697 from shaken flasks to a stirred-tank bioreactor with the optimized media and fructose as carbon source. The second purpose is to improve the production of PHB by applying both the batch and fed-batch fermentation strategies in comparison with previous works of wild type C. necator with fructose. Furthermore, thinking of biomedical applications, physicochemical, and cytotoxicity analyses of the produced biopolymer, are presented. Fed-batch fermentation with an exponential feeding strategy enabled us to achieve the highest values of PHB concentration and productivity, 25.7 g/l and 0.43 g/(l h), respectively. The PHB productivity was 3.3 and 7.2 times higher than the one in batch strategy and shaken flask cultures, respectively. DSC, FTIR, 1 H, and 13 C NMR analysis led to determine that the biopolymer produced by C. necator ATCC 17697 has a molecular structure and characteristics in agreement with the commercial PHB. Additionally, the biopolymer does not induce cytotoxic effects on the NIH/3T3 cell culture. Due to the improved fermentation strategies, PHB concentration resulted in 40 % higher of the already reported one for wild type C. necator using other fed-batch modes and fructose as a carbon source. Thus the produced PHB could be attractive for biomedical applications, which generate a rising interest in polyhydroxyalkanoates during recent years.

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“…At 235nm, the alpha, beta-unsaturated acids, and beta-hydroxy acids converted into crotonic acid on treatment with conc.H 2 SO 4 ; this is a rapid UV spectrophotometric assay reported so far [ 30 ]. Likewise, crotonic acid is estimated in the production of short side chain-polyhydroxyalkanoates from the Ralstonia pickettii [ 31 ]. Cell culture was cultivated as reported previously, and the cell pellet was dried to measure the dry weight of the cell (DCW) in g/L.…”

Section: Discussionmentioning

confidence: 99%

Selective isolation and genomic characterization of biopolymer producer—a novel feature of halophile Brachybacterium paraconglomeratum MTCC 13074

Mandragutti

Gódi

2023

Journal of Genetic Engineering and Biotechnology

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Background Biopolymers like polyhydroxyalkanoates (PHA) are the best natural macromolecules to use as alternative to the synthetic polymers. Many prokaryotes accumulate PHA as cytoplasmic intracellular granules and their accumulation is triggered by starving conditions. The PHAs are ecofriendly and used to create biodegradable plastics. The microbial synthesized PHA had acquired global importance in industrial and biomedical sectors. Results Ten different bacterial strains were isolated for the screening of PHA producers from the estuarine region of the Bay of Bengal, Suryalanka in Bapatla. A yellowish slimy circular colony known as M4 is actively growing on selective minimal media and was screened for polymeric granules in its cytoplasm using Sudan Black B and confirmed with the fluorescent dye Nile blue A. All of the isolates were biochemically tested and isolate M4 is the most capable of growing at high NaCl concentrations (3.2 percent) and tests positive for catalase, methyl red. The M4 strain revealed clear hydrolysis of gelatin, starch, and casein. The 16S rRNA sequencing revealed that M4 is 99.72% of identity to Brachybacterium paraconglomeratum LMG 19861(T) in BLAST and the obtained strain was assigned with accession no. MTCC 13074 and deposited in NCBI with accession no. MW899045. The chief cellular fatty acids found in M4 were C14:0, C15:0, C16:0, C18:1cis-9, C18:0, iso-C15: 0, iso-C14: 0, anteiso-C17: 0 and C18:1-7. Crotonic acid formation from M4-PHB extract was detected at 235nm in a UV spectrophotometer. Methanolysis was done, and derivatives of polyhydroxybutyric acid (PHB) in the extract were analyzed using GC-MS. Increasing viscosity was seen in the extracts which confirms the presence of polymer in the extracts. Thermogravimetric analysis was studied to determine the thermal profile of the PHB in the extract of M4. Conclusion In the study, the selective screening and extraction of ecofriendly PHB from M4 strain was highlighted. Brachybacterium paraconglomeratum is a novel strain showed its uniqueness by producing few monomeric derivatives of PHB. The strain was reporting for the first time as PHA producer. B. paraconglomeratum has promising characteristics according to its metabolic profile. In addition, this study also helps to understand the diversity of bacteria isolated from marine sources.

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Production of Short Side Chain-Poly[Hydroxyalkanoate] by a Newly Isolated Ralstonia Pickettii Strain

Cited by 19 publications

References 40 publications

In situ protein‐SIP highlights Burkholderiaceae as key players degrading toluene by para ring hydroxylation in a constructed wetland model

In situ protein‐SIP highlights Burkholderiaceae as key players degrading toluene by para ring hydroxylation in a constructed wetland model

Improved fermentation strategies in a bioreactor for enhancing poly(3-hydroxybutyrate) (PHB) production by wild type Cupriavidus necator from fructose

Selective isolation and genomic characterization of biopolymer producer—a novel feature of halophile Brachybacterium paraconglomeratum MTCC 13074

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