Isolation and Characterization of a Burkholderia sp. USM (JCM15050) Capable of Producing Polyhydroxyalkanoate (PHA) from Triglycerides, Fatty Acids and Glycerols

Journal of Polymers and the Environment (Impact Factor: 1.67). 12/2010; 18(4):584-592. DOI: 10.1007/s10924-010-0204-1


A consortium of microorganisms from oil polluted wastewater sample was cultivated to promote polyhydroxyalkanoate (PHA) accumulation
before subjecting the mixed cultures to sucrose density gradient ultracentrifugation. This resulted in the fractionation of
the bacterial cells according to their physical features such as size, morphology and/or densities. An isolate was identified
as Burkholderia sp. USM (JCM15050), which was capable of converting palm oil products [crude palm kernel oil (CPKO), palm olein (PO), palm
kernel acid oil (PKAO), palm stearin (PS), crude palm oil (CPO), palm acid oil (PAO) and palm fatty acid distillate (PFAD)],
fatty acids and various glycerol by-products into poly(3-hydroxybutyrate) [P(3HB)]. Up to 70 and 60wt% of P(3HB) could be
obtained when 0.5%(v/v) CPKO and glycerol was fed, respectively. Among the various fatty acids tested, lauric acid followed
by oleic acid and myristic acid gave the best cell growth and PHA accumulation. Compared to Cupriavidus necator H16, the present isolate showed better ability to grow on and produce PHA from various glycerol by-products generated by
the palm oil industry. This study demonstrated for the first time an isolate that has the potential to utilize palm oil and
glycerol derivatives for the biosynthesis of PHA.

KeywordsPHA-Ultracentrifugation-Palm oil-Glycerol-
Burkholderia sp.

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Available from: Jiun Yee Chee
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    • "However, the cells begin to increase in size and weight due to the intracellular accumulation of PHAs as a storage product (Lee, 1996). Fed-batch cultivation is more efficient than batch cultivation for achieving high product-and cell-concentration because it is possible to avoid inhibition by overly high concentrations of the substrate (Chee et al., 2010a). Single fed-batch fermentations that are nitrogen limited lead to low amounts of polymer, because there is not enough accumulation of biomass (Katircioglu et al., 2003), thus two-stage fed-batch cultivation is usually employed (Diniz et al., 2004). "
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    ABSTRACT: The development of processes for the production of biopolymer materials is being stimulated by a combination of factors. These factors include the negative effects of petrochemical-derived plastics on the global environment, depletion of global fossil fuel supplies, and the growing demands of an ever-increasing population for the products deemed necessary for an affluent modern lifestyle. In particular, polyhydroxyalkanoates have attracted attention as environmentally friendly alternatives to the synthetic polymers that are commonly used. Polyhydroxyalkanoates are polyesters produced and accumulated in intracellular granules by many microorganisms. Because they are biodegradable and biocompatible and can be produced by fermentation of renewable feedstocks, they are considered attractive substitutes for petroleum-derived polymers. To create bacterial polyesters, crude and waste plant oils, which can be difficult to dispose of, can be recovered and used as feedstock. This paper gives an overview of the potential for the production of polyhydroxyalkanoates with useful physicochemical properties by bacteria grown on renewable resources such as plant oils.
    Full-text · Article · Oct 2015 · Journal of Cleaner Production
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    • "A large group of microorganisms is capable of assimilating glycerol as a carbon source in the synthesis of many useful products, such as 1,3-PD, ethanol, hydrogen, polyhydroxyalkanoates, and organic acids (Amaral et al. 2009; da Silva et al. 2009; André et al. 2010; Chatzifragkou et al. 2011a, b, c). A number of microorganisms can grow on glycerol, including Actinobacillus succinogenes, Aspergillus niger, Blakeslea trispora, Burkholderia sp., Chlorella protothecoides , Citrobacter freundii , Clostridium buturicum , Clostridium pasteurianum , Cunninghamella echinulata, Cupriavidus necator, Enterobacter aerogenes , Escherichia coli, Gluconobacter sp., Klebsiella pneumoniae, Kluyvera cryocrescens , Lentinula edodes , Mortierella ramanniana , Mucor sp., Pseudomonas oleovorans , Rhodotorula glutinis, Schizochytrium limacinum, Staphylococcus caseolyticus , Yarrowia lipolytica , and Zobellella denitrificans (Petitdemange et al. 1995; González-Pajuelo et al. 2004; Hirschmann et al. 2005; Ito et al. 2005; Mu et al. 2006; Rymowicz et al. 2006, 2009; Fakas et al. 2008; Mantzouridou et al. 2008; Volpato et al. 2008; Cavalheiro et al. 2009; Habe et al. 2009; Rywińska et al. 2009; André et al. 2010; Andreeßen et al. 2010; Chatzifragkou et al. 2010; Chee et al. 2010; Ibrahim and Steinbüchel 2010; Liang et al. 2010; Ashby et al. 2011; Choi et al. 2011; O'Grady and Morgan 2011; Saenge et al. 2011; Vlysidis et al. 2011; Bellou et al. 2012; Metsoviti et al. 2012; Venkataramanan et al. 2012; Wilkens et al. 2012). Table 1 presents a number of the microorganisms that are able to convert crude glycerol to commercially useful metabolites as well as transform the main impurities of this raw material.. "
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    ABSTRACT: Glycerol is a valuable raw material for the production of industrially useful metabolites. Among many promising applications for the use of glycerol is its bioconversion to high value-added compounds, such as 1,3-propanediol (1,3-PD), succinate, ethanol, propionate, and hydrogen, through microbial fermentation. Another method of waste material utilization is the application of crude glycerol in blends with other wastes (e.g., tomato waste hydrolysate). However, crude glycerol, a by-product of biodiesel production, has many impurities which can limit the yield of metabolites. In this mini-review we summarize the effects of crude glycerol impurities on various microbial fermentations and give an overview of the metabolites that can be synthesized by a number of prokaryotic and eukaryotic microorganisms when cultivated on glycerol.
    Full-text · Article · Sep 2014 · Annals of Microbiology
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    • "This is a very interesting carbon source for certain microbes. Glycerol can be used to produce PHB from Burkholderia sp (Chee et al., 2012; Zhu et al., 2010) "
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    ABSTRACT: Conventional plastics, though widely used have a number of disadvantages like the dependency on depleting fossil fuels for their production, the long time required for decomposition of the material and the associated production of toxic materials. Bio-based plastics are a solution to these problems. Among the many types of Bioplastics, Polyhydroxyalkanoates (PHAs) have been studied widely because of their similarity with conventional plastics and complete biodegradability. Polyhydroxyalkanoates are produced in microorganisms as intracellular storage compounds of energy and carbon under conditions of non-carbon nutrient deficiency.These polymers are generally classified in two categories depending on the number of carbon atoms in their monomer units: small chain length (scl)-PHA when the monomer units contain from 3 to 5 carbon atoms and medium chain length (mcl)-PHA with monomer units possessing from 6 to14 carbon atoms. PHAs, owing to their lipid nature are insoluble in water. The polymers are accumulated as intracellular granules bound by a layer of Phospholipids and some associated proteins. This kind of storage helps in accumulation of large amounts of carbon without disturbing the osmotic balance of the cell. The polymer can be degraded completely using PHA hydrolases and PHA depolymerases thus making it completely compatible with the Carbon cycle and therefore pose no threats to the environment like the conventional plastics. The only limitation in the widespread use of the polymer is the high cost of production in comparison to the conventional plastics. Research on usage of cheaper substrates is being investigated currently to bring down the cost of production. PHAs have been developed to suit a number of applications including the medical field. The chapter studies the properties of PHA, the cost cutting strategies that can be incorporated for large scale production of the polymer and the benefits of using PHA over conventional plastics.
    Full-text · Chapter · Aug 2014
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