Removal of Endotoxin during Purification of Poly(3-Hydroxybutyrate) from Gram-Negative Bacteria

Lee, Sang Yup; Choi, Jong‐il; Han, Koeun; Song, Ji Yong

doi:10.1128/aem.65.6.2762-2764.1999

Cited by 102 publications

(41 citation statements)

References 21 publications

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“…The presence of LPS induces a strong immunogenic reaction and is therefore undesirable for the biomedical application of the PHAs (Chen and Wu 2005;Valappil et al 2006). Gram-positive bacteria lack LPS and are hence better sources of PHAs for use in biomedical applications (Lee et al 1999;Valappil et al 2007a). However, the presence of lipidated macroamphiphiles in PHAs isolated from this group of bacteria needs to be investigated (Valappil et al 2007a).…”

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confidence: 99%

Polyhydroxyalkanoate biosynthesis in Bacillus cereus SPV under varied limiting conditions and an insight into the biosynthetic genes involved

et al. 2008

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Aims: A new strain of Bacillus, Bacillus cereus SPV, was found to be capable of using a wide range of carbon sources for the production of polyhydroxyalkanoates (PHAs) (Valappil et al. 2007b). Limiting nutrient in the culture conditions is crucial for PHA production. In this study, B. cereus SPV was grown in different culture conditions with limitation of potassium, nitrogen, sulphur and phosphorous to establish the impact of nutritional limitation on PHA production. Methods and Results: The PHA yields obtained were found to be 13·4, 38, 13·15 and 33·33% dcw for potassium, nitrogen, sulphur and phosphorus limitations, respectively. Gas chromatography–mass spectrometry analysis of the isolated polymers showed the presence of P(3HB) under nitrogen, sulphur and phosphate‐limiting conditions and P(3HB‐3HV) copolymer under potassium limiting conditions. This ability of B. cereus SPV to accumulate different PHA monomers from structurally unrelated carbon sources led to an interest in the molecular analysis of PHA biosynthesis in this organism. To achieve this, PCR was used to identify the polyhydroxyalkanoate biosynthetic genes in B. cereus SPV. Conclusion: Sequence analysis of the PCR products from B. cereus SPV revealed the sequence of the putative biosynthetic genes, and possible regions involved in substrate binding. The nucleotide sequence reported in this paper is in the GenBank nucleotide sequence database under accession number . Significance and Impact of the Study: This is the first report comparing the capability of B. cereus SPV to produce PHAs under different culture conditions of potassium, nitrogen, sulfur and phosphate limitations. The results in this study suggest the unique ability of B. cereus SPV to supply both 3HB and 3HV monomers from a structurally unrelated carbon source, glucose.

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Polyhydroxyalkanoate biosynthesis in Bacillus cereus SPV under varied limiting conditions and an insight into the biosynthetic genes involved

et al. 2008

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“…The concentration of endotoxins in PHA is greatly influenced by purification strategy and might vary from more than 10 4 EU/g to less than 1 EU/g. 55,98 The methodology for endotoxin elimination depends on type of PHA (e.g., scl-PHA, mcl-PHA, presence of functional groups, etc.) and each results in different rates of polymer recovery.…”

Section: Bug Systems For Scaling Up: Wild Type Over Recombinant Cellsmentioning

confidence: 99%

“…and each results in different rates of polymer recovery. 55,98 However, in vitro strategy remains hampered by the necessity of extensive and tedious purification methodology to achieve the levels in compliance with the endotoxin requirements for biomedical application according to the U.S. Food and Drug Administration (FDA). Generally, for products that directly or indirectly contact the cardiovascular system and lymphatic system the limit is 0.5 EU/mL or 20 EU/ device, while for devices in contact with cerebrospinal fluid the limit is 0.06 EU/mL or 2.15 EU/device.…”

Section: Bug Systems For Scaling Up: Wild Type Over Recombinant Cellsmentioning

confidence: 99%

“…105,106 The genus Bacillus, in common with many other PHA-accumulating Gram-positive bacteria, accumulates co-polymers of 3HB when grown on different substrates. 98 For instance, copolymers of P(3HB-co-3HV) are accumulated when the cultures are fed with odd-chain-length n-alkanoic acids such as propionic acid, valeric acid and heptanoic acid. 107 The generally-regarded-as-safe (GRAS) bacterium Lactococcus lactis has been genetically engineered to produce PHA beads.…”

Section: Bug Systems For Scaling Up: Wild Type Over Recombinant Cellsmentioning

confidence: 99%

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Smart polyhydroxyalkanoate nanobeads by protein based functionalization

Dinjaski

Prieto

2015

Nanomedicine: Nanotechnology, Biology and Medicine

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In the 21st century, we are coming into the age of personalized medicine. There is a growing use of biomaterials in the clinical setting. In this review article, the authors describe the use of natural polyhydroxyalkanoate (PHA) nanoparticulates, which are formed within bacterial cells and can be easily functionalized. The potential uses would include high-affinity bioseparation, enzyme immobilization, protein delivery, diagnostics etc. The challenges of this approach remain the possible toxicity from endotoxin and the high cost of production.

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“…The most commonly used solvents for this purpose are chlorinated hydrocarbons such as chloroform that is, however, too toxic, expensive, and nonrecyclable. One way to minimize these disadvantages is to replace the chloroform with a cheap and less toxic alternative in the course of PHB extraction [8][9][10].…”

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confidence: 99%

Comparison of different solvents for extraction of polyhydroxybutyrate from Cupriavidus necator

Aramvash

Zavareh

Banadkuki

2017

Engineering in Life Sciences

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Polyhydroxybutyrate (PHBs) have attracted much attention due to their biodegradability and biocompatibility properties. The main drawback to the commercial production of them is their high cost. The recovery of PHB from bacterial cytoplasm significantly increases total processing costs. Efficient, economical, and environmentfriendly extraction of PHB from cells is required for its industrial production. In the present study, several nonhalogenated organic solvents (ethylene carbonate, dimethyl sulfoxide, dimethyl formamide, hexane, propanol, methanol, and acetic acid) were examined for their efficacy regarding recovery at different temperatures from culture broth containing Cupriavidus necator cells. The highest recovery percentage (98.6%) and product purity (up to 98%) were seen to be those of ethylene carbonate-assisted extraction at 150°C within 60 min of incubation time. Average molecular weight of the recovered PHB (1.3 × 10 6 ) was not significantly affected by the extraction solvent and conditions. The melting point of PHB extracted using ethylene carbonate was measured to be 176.2°C with an enthalpy of fusion of 16.8% and the corresponding degree of crystallinity of 59.2%. NMR and GC analyses confirmed that the extracted biopolymer was PHB. The presented strategy can help researchers to reduce the cost to obtain the final product.

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Removal of Endotoxin during Purification of Poly(3-Hydroxybutyrate) from Gram-Negative Bacteria

Cited by 102 publications

References 21 publications

Polyhydroxyalkanoate biosynthesis in Bacillus cereus SPV under varied limiting conditions and an insight into the biosynthetic genes involved

Polyhydroxyalkanoate biosynthesis in Bacillus cereus SPV under varied limiting conditions and an insight into the biosynthetic genes involved

Smart polyhydroxyalkanoate nanobeads by protein based functionalization

Comparison of different solvents for extraction of polyhydroxybutyrate from Cupriavidus necator

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