Displaying publications 41 - 60 of 82 in total

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  1. The oil-absorbing property of polyhydroxyalkanoate films and its practical application: a refreshing new outlook for an old degrading material
    Sudesh K, Loo CY, Goh LK, Iwata T, Maeda M
    Macromol Biosci, 2007 Nov 12;7(11):1199-205.
    PMID: 17703476
    Polyhydroxyalkanoates (PHAs) have attracted the attention of academia and industry because of their plastic-like properties and biodegradability. However, practical applications as a commodity material have not materialized because of their high production cost and unsatisfactory mechanical properties. PHAs are also believed to have high-value applications as an absorbable biomaterial for tissue engineering and drug-delivery devices because of their biocompatibility. However, research in these areas is still in its very early stages. The main problem faced by proponents of PHAs is the lack of a niche area where PHAs will be the most desired material in terms of its function during use rather than because of its eco-friendly virtues after use. Here, we report on the oil-absorbing property of PHA films and its potential applications. By comparing with some of the existing commercial products, the potential application of PHAs as cosmetic oil-blotting films is revealed for the first time. Besides having the ability to rapidly absorb and retain oil, PHA films also have a natural oil-indicator property, showing obvious changes in opacity following oil absorption. Surface analysis revealed that the surface structures such as porosity and smoothness exert great influence on the rapid oil-absorption properties of the PHA films. These newly discovered properties could be exploited to create a niche area for the practical applications of PHAs.
    Matched MeSH terms: Polyhydroxyalkanoates/pharmacokinetics*; Polyhydroxyalkanoates/chemistry*
  2. Effects of even and odd number fatty acids cofeeding on PHA production and composition in Pseudomonas putida Bet001 isolated from palm oil mill effluent
    Mohd Razaif-Mazinah MR, Mohamad Annuar MS, Sharifuddin Y
    Biotechnol Appl Biochem, 2016 Jan-Feb;63(1):92-100.
    PMID: 25643814 DOI: 10.1002/bab.1354
    The biosynthesis of medium-chain-length poly-3-hydroxyalkanoates by Pseudomonas putida Bet001 cultivated on mixed carbon sources was investigated. The mixed carbon sources consisted of heptanoic acid (HA) and oleic acid (OA). A relatively low PHA content at 1.2% (w/w) and 11.4% (w/w) was obtained when HA or OA was used as the sole carbon source. When these fatty acids were supplied as a mixture, PHA content increased threefold. Interestingly, the mixture-derived PHA composed of both odd and even monomer units, namely. 3-hydroxyheptanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, and 3-hydroxydodecanoate and no unsaturated monomer was detected. It is hypothesized that the even-numbered monomers were derived primarily from OA, whereas the odd-numbered monomer was derived from HA. This also points out to an efficient and yet distinct fatty acids metabolism that fed the PHA biosynthesis machinery of this particular microorganism. PHA obtained was elastomeric because melting temperature (Tm ) and crystallinity were absent. It showed good thermal stability with degradation temperature (Td ) ranging from 275.96 to 283.05 °C.
    Matched MeSH terms: Polyhydroxyalkanoates/metabolism*; Polyhydroxyalkanoates/chemistry
  3. Enhancement of stress tolerance in the polyhydroxyalkanoate producers without mobilization of the accumulated granules
    Goh LK, Purama RK, Sudesh K
    Appl Biochem Biotechnol, 2014 Feb;172(3):1585-98.
    PMID: 24233544 DOI: 10.1007/s12010-013-0634-z
    Poly(3-hydroxybutyrate) [P(3HB)], a polymer belonging to the polyhydroxyalkanoate (PHA) family, is accumulated by numerous bacteria as carbon and energy storage material. The mobilization of accumulated P(3HB) is associated with increased stress and starvation tolerance. However, the potential function of accumulated copolymer such as poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] remained unknown. In this study, Delftia acidovorans DS 17 was used to evaluate the contributions of P(3HB) and P(3HB-co-3HV) granules during simulated exogenous carbon deprivation on cell survival by transferring cells with PHAs to carbon-free mineral salt medium supplemented with 1% (w/v) nitrogen source. By mobilizing the intracellular P(3HB) and P(3HB-co-3HV) at 11 and 40 mol% 3HV compositions, the cells survived starvation. Surprisingly, D. acidovorans containing P(3HB-co-94 mol% 3HV) also survived although the mobilization was not as effective. Similarly, recombinant Escherichia coli pGEM-T::phbCAB(Cn) (harboring the PHA biosynthesis genes of Cupriavidus necator) containing P(3HB) granules had a higher viable cell counts compared to those without P(3HB) granules but without any P(3HB) mobilization when exposed to oxidative stress by photoactivated titanium dioxide. This study provided strong evidence that enhancement of stress tolerance in PHA producers can be achieved without mobilization of the previously accumulated granules. Instead, PHA biosynthesis may improve bacterial survival via multiple mechanisms.
    Matched MeSH terms: Polyhydroxyalkanoates/biosynthesis*; Polyhydroxyalkanoates/chemistry
  4. Fulltext Biosynthesis and characterization of polyhydroxyalkanoates copolymers produced by Pseudomonas putida Bet001 isolated from palm oil mill effluent
    Gumel AM, Annuar MS, Heidelberg T
    PLoS One, 2012;7(9):e45214.
    PMID: 23028854 DOI: 10.1371/journal.pone.0045214
    The biosynthesis and characterization of medium chain length poly-3-hydroxyalkanoates (mcl-PHA) produced by Pseudomonas putida Bet001 isolated from palm oil mill effluent was studied. The biosynthesis of mcl-PHA in this newly isolated microorganism follows a growth-associated trend. Mcl-PHA accumulation ranging from 49.7 to 68.9% on cell dry weight (CDW) basis were observed when fatty acids ranging from octanoic acid (C(8:0)) to oleic acid (C(18:1)) were used as sole carbon and energy source. Molecular weight of the polymer was found to be ranging from 55.7 to 77.7 kDa. Depending on the type of fatty acid used, the (1)H NMR and GCMSMS analyses of the chiral polymer showed a composition of even and odd carbon atom chain with monomer length of C4 to C14 with C8 and C10 as the principal monomers. No unsaturated monomer was detected. Thermo-chemical analyses showed the accumulated PHA to be semi-crystalline polymer with good thermal stability, having a thermal degradation temperature (T(d)) of 264.6 to 318.8 (± 0.2) (o)C, melting temperature (T(m)) of 43. (± 0.2) (o)C, glass transition temperature (T(g)) of -1.0 (± 0.2) (o)C and apparent melting enthalpy of fusion (ΔH(f)) of 100.9 (± 0.1) J g(-1).
    Matched MeSH terms: Polyhydroxyalkanoates/biosynthesis*; Polyhydroxyalkanoates/isolation & purification*
  5. Development of polyhydroxyalkanoates production from waste feedstocks and applications
    Pakalapati H, Chang CK, Show PL, Arumugasamy SK, Lan JC
    J Biosci Bioeng, 2018 May 23.
    PMID: 29803402 DOI: 10.1016/j.jbiosc.2018.03.016
    Polyhydroxyalkanoates (PHA) are naturally occurring biopolymers, obtained from microorganisms. Properties like biodegradability and biocompatibility make PHA a part of today's commercial polymer industry. However, the production cost of PHA has been a great barrier to extend its application to large scale production. Substrates and usage of pure cultures constitute the main reason for its high production cost. On the other hand, rapid industrialization i.e., industrial sectors such as sugar, pulp and paper, fruit and food processing, dairies, slaughterhouses, and poultries, has resulted in the generation of the huge quantity of wastes. Consequently, becoming large source of environmental pollution and health hazard. This review emphasizes on the usage of various waste feedstocks obtained from industrial and agricultural industries as an alternate substrate for PHA production. As these waste materials are rich in organic material and also microbes, they can be the good starting material for PHA production. Additionally, advantages and economic importance of mixed cultures and also PHA applications are discussed. Future prospects and challenges in PHA production from waste feedstocks are also highlighted.
    Matched MeSH terms: Polyhydroxyalkanoates
  6. Application of thermo-separating aqueous two-phase system in extractive bioconversion of polyhydroxyalkanoates by Cupriavidus necator H16
    Leong YK, Show PL, Lan JC, Krishnamoorthy R, Chu DT, Nagarajan D, et al.
    Bioresour Technol, 2019 Sep;287:121474.
    PMID: 31122870 DOI: 10.1016/j.biortech.2019.121474
    Polyhydroxyalkanoates (PHAs), a family of biodegradable and renewable biopolymers show a huge potential as an alternative to conventional plastics. Extractive bioconversion (in situ product recovery) is a technique that integrates upstream fermentation and downstream purification. In this study, extractive bioconversion of PHAs from Cupriavidus necator H16 was performed via a thermo-separating aqueous two-phase system to reduce the cost and environmental impacts of PHAs production. Key operating parameters, such as polymer concentration, temperature, and pH, were optimized. The strategy achieved a yield and PF of 97.6% and 1.36-fold, respectively at 5% EOPO 3900 concentration, 30 °C fermentation temperature and pH 6. The PHAs production process was also successfully scaled up in a 2 L bioreactor. To the best of our knowledge, this is the first report on extractive fermentation of PHAs from Cupriavidus necator utilizing a thermo-separation system to achieve a better productivity and purity of the target product.
    Matched MeSH terms: Polyhydroxyalkanoates
  7. Fulltext Dataset on controlled production of polyhydroxyalkanoate-based microbead using double emulsion solvent evaporation technique
    Govindasamy S, Syafiq IM, Amirul AA, Amin RM, Bhubalan K
    Data Brief, 2019 Apr;23:103675.
    PMID: 30788397 DOI: 10.1016/j.dib.2019.01.023
    A significant source of microplastics is from the usage of microbeads in the market since petrochemical plastic bead is a material used in cosmetic scrubs. A possible way to counteract the problem is by the substitution of synthetic plastic to natural biodegradable polymer. Polyhydroxyalkanoate (PHA) is a general class of thermoplastic microbial polymer and it is the best alternative to some petrochemical plastics due to its biodegradability. Some PHA has earned its way into cosmetic application due to its biocompatibility. This data article reports data on the development of biodegradable microbeads by using the double emulsion solvent evaporation technique. Our data describe the extraction of biopolymer from marine bacteria that was cultivated in shaken flask culture, removal of endotoxins using oxidizing agent, the production of microbeads using a peristaltic pump with a specific flowrate and silicon tubing, and the cytotoxicity of the microbeads.
    Matched MeSH terms: Polyhydroxyalkanoates
  8. Fulltext Statistical Design of Experimental and Bootstrap Neural Network Modelling Approach for Thermoseparating Aqueous Two-Phase Extraction of Polyhydroxyalkanoates
    Leong YK, Chang CK, Arumugasamy SK, Lan JC, Loh HS, Muhammad D, et al.
    Polymers (Basel), 2018 Jan 30;10(2).
    PMID: 30966168 DOI: 10.3390/polym10020132
    At present, polyhydroxyalkanoates (PHAs) have been considered as a promising alternative to conventional plastics due to their diverse variability in structure and rapid biodegradation. To ensure cost competitiveness in the market, thermoseparating aqueous two-phase extraction (ATPE) with the advantages of being mild and environmental-friendly was suggested as the primary isolation and purification tool for PHAs. Utilizing two-level full factorial design, this work studied the influence and interaction between four independent variables on the partitioning behavior of PHAs. Based on the experimental results, feed forward neural network (FFNN) was used to develop an empirical model of PHAs based on the ATPE thermoseparating input-output parameter. In this case, bootstrap resampling technique was used to generate more data. At the conditions of 15 wt % phosphate salt, 18 wt % ethylene oxide⁻propylene oxide (EOPO), and pH 10 without the addition of NaCl, the purification and recovery of PHAs achieved a highest yield of 93.9%. Overall, the statistical analysis demonstrated that the phosphate concentration and thermoseparating polymer concentration were the most significant parameters due to their individual influence and synergistic interaction between them on all the response variables. The final results of the FFNN model showed the ability of the model to seamlessly generalize the relationship between the input⁻output of the process.
    Matched MeSH terms: Polyhydroxyalkanoates
  9. Fulltext Asymmetric Open-Closed Dimer Mechanism of Polyhydroxyalkanoate Synthase PhaC
    Chek MF, Kim SY, Mori T, Tan HT, Sudesh K, Hakoshima T
    iScience, 2020 May 22;23(5):101084.
    PMID: 32388399 DOI: 10.1016/j.isci.2020.101084
    Biodegradable polyester polyhydroxyalkanoate (PHA) is a promising bioplastic material for industrial use as a replacement for petroleum-based plastics. PHA synthase PhaC forms an active dimer to polymerize acyl moieties from the substrate acyl-coenzyme A (CoA) into PHA polymers. Here we present the crystal structure of the catalytic domain of PhaC from Chromobacterium sp. USM2, bound to CoA. The structure reveals an asymmetric dimer, in which one protomer adopts an open conformation bound to CoA, whereas the other adopts a closed conformation in a CoA-free form. The open conformation is stabilized by the asymmetric dimerization and enables PhaC to accommodate CoA and also to create the product egress path. The bound CoA molecule has its β-mercaptoethanolamine moiety extended into the active site with the terminal SH group close to active center Cys291, enabling formation of the reaction intermediate by acylation of Cys291.
    Matched MeSH terms: Polyhydroxyalkanoates
  10. Fulltext A Review of the Applications and Biodegradation of Polyhydroxyalkanoates and Poly(lactic acid) and Its Composites
    Boey JY, Mohamad L, Khok YS, Tay GS, Baidurah S
    Polymers (Basel), 2021 May 12;13(10).
    PMID: 34065779 DOI: 10.3390/polym13101544
    Overconsumption of plastic goods and improper handling of petroleum-derived plastic waste have brought a plethora of negative impacts to the environment, ecosystem and human health due to its recalcitrance to degradation. These drawbacks become the main driving force behind finding biopolymers with the degradable properties. With the advancement in biopolymer research, polyhydroxyalkanoate (PHA) and poly(lacyic acid) (PLA) and its composites have been alluded to as a potential alternative to replace the petrochemical counterpart. This review highlights the current synthesis process and application of PHAs and PLA and its composites for food packaging materials and coatings. These biopolymers can be further ameliorated to enhance their applicability and are discussed by including the current commercially available packaging products. Factors influencing biodegradation are outlined in the latter part of this review. The main aim of this review article is to organize the scattered available information on various aspects of PHAs and PLA, and its composites for packaging application purposes. It is evident from a literature survey of about 140 recently published papers from the past 15 years that PLA and PHA show excellent physical properties as potential food packaging materials.
    Matched MeSH terms: Polyhydroxyalkanoates
  11. Fulltext Revelation of the ability of Burkholderia sp. USM (JCM 15050) PHA synthase to polymerize 4-hydroxybutyrate monomer
    Lau NS, Sudesh K
    AMB Express, 2012;2(1):41.
    PMID: 22877240 DOI: 10.1186/2191-0855-2-41
    The nutrition-versatility of Burkholderia sp. strain USM (JCM 15050) has initiated the studies on the use of this bacterium for polyhydroxyalkanoate (PHA) production. To date, the Burkholderia sp. has been reported to synthesize 3-hydroxybutyrate, 3-hydroxyvalerate and 3-hydroxy-4-methylvalerate monomers. In this study, the PHA biosynthetic genes of this strain were successfully cloned and characterized. The PHA biosynthetic cluster of this strain consisted of a PHA synthase (phaC), β-ketothiolase (phaA), acetoacetyl-CoA reductase (phaB) and PHA synthesis regulator (phaR). The translated products of these genes revealed identities to corresponding proteins of Burkholderia vietnamiensis (99-100 %) and Cupriavidus necator H16 (63-89%). Heterologous expression of phaCBs conferred PHA synthesis to the PHA-negative Cupriavidus necator PHB¯4, confirming that phaCBs encoded functionally active protein. PHA synthase activity measurements revealed that the crude extracts of C. necator PHB¯4 transformant showed higher synthase activity (243 U/g) compared to that of wild-types Burkholderia sp. (151 U/g) and C. necator H16 (180 U/g). Interestingly, the transformant C. necator PHB¯4 harbouring Burkholderia sp. PHA synthase gene accumulated poly(3-hydroxybutyrate-co-4-hydroxybutyrate) with 4-hydroxybutyrate monomer as high as up to 87 mol% from sodium 4-hydroxybutyrate. The wild type Burkholderia sp. did not have the ability to produce this copolymer.
    Matched MeSH terms: Polyhydroxyalkanoates
  12. Fulltext Densitometry based microassay for the determination of lipase depolymerizing activity on polyhydroxyalkanoate
    Ch'ng DH, Sudesh K
    AMB Express, 2013;3(1):22.
    PMID: 23657221 DOI: 10.1186/2191-0855-3-22
    A novel method for the assay of polyhydroxyalkanoate (PHA)-degrading ability of triacylglycerol lipases was developed. By applying the natural affinity of lipases towards hydrophobic interfaces, a sensitive and rapid densitometry analysis for the evaluation of hydrolytic activity of lipase droplets towards PHA-coated surface was successfully carried out. We found that 12 out of 14 tested lipases which are of fungal, bacterial and animal origin were able to hydrolyze P(3HB-co-92 mol% 4HB) thin film. The patterns and opacity of the hydrolysis spots of lipases on PHA films allowed easy comparison of PHA-hydrolytic strength of lipases. Lipase from the bacterium Chromobacterium viscosum exhibited the highest PHA-degrading activity. The hydrolytic activity of lipases on water insoluble PHA, emulsified p-nitrophenyl laurate and olive oil were also compared and interestingly some lipases showed better activity when PHA was used as a substrate.
    Matched MeSH terms: Polyhydroxyalkanoates
  13. Discovery of a new polyhydroxyalkanoate synthase from limestone soil through metagenomic approach
    Tai YT, Foong CP, Najimudin N, Sudesh K
    J Biosci Bioeng, 2016 Apr;121(4):355-64.
    PMID: 26467694 DOI: 10.1016/j.jbiosc.2015.08.008
    PHA synthase (PhaC) is the key enzyme in the production of biodegradable plastics known as polyhydroxyalkanoate (PHA). Nevertheless, most of these enzymes are isolated from cultivable bacteria using traditional isolation method. Most of the microorganisms found in nature could not be successfully cultivated due to the lack of knowledge on their growth conditions. In this study, a culture-independent approach was applied. The presence of phaC genes in limestone soil was screened using primers targeting the class I and II PHA synthases. Based on the partial gene sequences, a total of 19 gene clusters have been identified and 7 clones were selected for full length amplification through genome walking. The complete phaC gene sequence of one of the clones (SC8) was obtained and it revealed 81% nucleotide identity to the PHA synthase gene of Chromobacterium violaceum ATCC 12472. This gene obtained from uncultured bacterium was successfully cloned and expressed in a Cupriavidus necator PHB(-)4 PHA-negative mutant resulting in the accumulation of significant amount of PHA. The PHA synthase activity of this transformant was 64 ± 12 U/g proteins. This paper presents a pioneering study on the discovery of phaC in a limestone area using metagenomic approach. Through this study, a new functional phaC was discovered from uncultured bacterium. Phylogenetic classification for all the phaCs isolated from this study has revealed that limestone hill harbors a great diversity of PhaCs with activities that have not yet been investigated.
    Matched MeSH terms: Polyhydroxyalkanoates
  14. A novel biological recovery approach for PHA employing selective digestion of bacterial biomass in animals
    Ong SY, Zainab-L I, Pyary S, Sudesh K
    Appl Microbiol Biotechnol, 2018 Mar;102(5):2117-2127.
    PMID: 29404644 DOI: 10.1007/s00253-018-8788-9
    Polyhydroxyalkanoate (PHA) is a family of microbial polyesters that is completely biodegradable and possesses the mechanical and thermal properties of some commonly used petrochemical-based plastics. Therefore, PHA is attractive as a biodegradable thermoplastic. It has always been a challenge to commercialize PHA due to the high cost involved in the biosynthesis of PHA via bacterial fermentation and the subsequent purification of the synthesized PHA from bacterial cells. Innovative enterprise by researchers from various disciplines over several decades successfully reduced the cost of PHA production through the efficient use of cheap and renewable feedstock, precisely controlled fermentation process, and customized bacterial strains. Despite the fact that PHA yields have been improved tremendously, the recovery and purification processes of PHA from bacterial cells remain exhaustive and require large amounts of water and high energy input besides some chemicals. In addition, the residual cell biomass ends up as waste that needs to be treated. We have found that some animals can readily feed on the dried bacterial cells that contain PHA granules. The digestive system of the animals is able to assimilate the bacterial cells but not the PHA granules which are excreted in the form of fecal pellets, thus resulting in partial recovery and purification of PHA. In this mini-review, we will discuss this new concept of biological recovery, the selection of the animal model for biological recovery, and the properties and possible applications of the biologically recovered PHA.
    Matched MeSH terms: Polyhydroxyalkanoates/metabolism*
  15. Exploration of Global Trend on Biomedical Application of Polyhydroxyalkanoate (PHA): A Patent Survey
    Paulraj P, Vnootheni N, Chandramohan M, Thevarkattil MJP
    Recent Pat Biotechnol, 2018;12(3):186-199.
    PMID: 29384069 DOI: 10.2174/1872208312666180131114125
    BACKGROUND: Polyhydroxyalkanoates are bio-based, biodegradable naturally occurring polymers produced by a wide range of organisms, from bacteria to higher mammals. The properties and biocompatibility of PHA make it possible for a wide spectrum of applications. In this context, we analyze the potential applications of PHA in biomedical science by exploring the global trend through the patent survey. The survey suggests that PHA is an attractive candidate in such a way that their applications are widely distributed in the medical industry, drug delivery system, dental material, tissue engineering, packaging material as well as other useful products.

    OBJECTIVE: In our present study, we explored patents associated with various biomedical applications of polyhydroxyalkanoates.

    METHOD: Patent databases of European Patent Office, United States Patent and Trademark Office and World Intellectual Property Organization were mined. We developed an intensive exploration approach to eliminate overlapping patents and sort out significant patents.We demarcated the keywords and search criterions and established search patterns for the database request. We retrieved documents within the recent 6 years, 2010 to 2016 and sort out the collected data stepwise to gather the most appropriate documents in patent families for further scrutiny.

    RESULTS: By this approach, we retrieved 23,368 patent documents from all the three databases and the patent titles were further analyzed for the relevance of polyhydroxyalkanoates in biomedical applications. This ensued in the documentation of approximately 226 significant patents associated with biomedical applications of polyhydroxyalkanoates and the information was classified into six major groups. Polyhydroxyalkanoates has been patented in such a way that their applications are widely distributed in the medical industry, drug delivery system, dental material, tissue engineering, packagingmaterial as well as other useful products.

    CONCLUSION: There are many avenues through which PHA & PHB could be used. Our analysis shows patent information can be used to identify various applications of PHA and its representatives in the biomedical field. Upcoming studies can focus on the application of PHA in the different field to discover the related topics and associate to this study.We believe that this approach of analysis and findings can initiate new researchers to undertake similar kind of studies in their represented field to fill the gap between the patent articles and research publications.

    Matched MeSH terms: Polyhydroxyalkanoates*
  16. Purification and characterization of new bio-plastic degrading enzyme from Burkholderia cepacia DP1
    Azura Azami N, Ira Aryani W, Aik-Hong T, Amirul AA
    Protein Expr Purif, 2019 03;155:35-42.
    PMID: 30352276 DOI: 10.1016/j.pep.2018.10.008
    Depolymerase is an enzyme that plays an important role in the hydrolysis of polyhydroxyalkanoates [PHAs]. In the current study, Burkholderia cepacia DP1 was obtained from Penang, Malaysia in which the enzyme was purified using ion exchange and gel filtration (Superdex-75) column chromatography. The molecular mass of the enzyme was estimated to be 53.3 kDa using SDS-PAGE. The enzyme activity was increased to 36.8 folds with the recovery of 16.3% after purification. The enzyme activity was detected between pH 6.0-10 and at 35-55 °C with pH 6.0 and 45 °C facilitating the maximum activity. Depolymerase was inactivated by Tween-20, Tween-80, SDS and PMSF, but insensitive to metal ions (Mg2+, Ca2+, K+, Na2+, Fe3+) and organic solvents (methanol, ethanol, and acetone). The apparent Km values of the purified P(3HB) depolymerase enzyme for P(3HB) and P(3HB-co-14%3HV) were 0.7 mg/ml and 0.8 mg/ml, respectively. The Vmax values of the purified enzyme were 10 mg/min and 8.89 mg/min for P(3HB) and P(3HB-co-14%3HV), respectively. The current study discovered a new extracellular poly(3-hydroxybutyrate) [P(3HB)] depolymerase enzyme from Burkholderia cepacia DP1 isolated and purified to homogeneity from the culture supernatant. To the best of our knowledge, this is the first report demonstrating the purification and biochemical characterization of P(3HB) depolymerase enzyme from genus Burkholderia.
    Matched MeSH terms: Polyhydroxyalkanoates/metabolism
  17. Complete Genome Sequence of a Novel Polyhydroxyalkanoate (PHA) Producer, Jeongeupia sp. USM3 (JCM 19920) and Characterization of Its PHA Synthases
    Zain NA, Ng LM, Foong CP, Tai YT, Nanthini J, Sudesh K
    Curr Microbiol, 2020 Mar;77(3):500-508.
    PMID: 31893298 DOI: 10.1007/s00284-019-01852-z
    A novel polyhydroxyalkanoate (PHA)-producing bacterium, Jeongeupia sp. USM3 (JCM 19920) was isolated from the limestone soil at Gua Tempurung, Perak, Malaysia. This is the first report on the complete genome sequence for the genus Jeongeupia. This genome consists of a circular chromosome with a size of 3,788,814 bp and contains 3557 genes. Two PHA synthase (phaC) genes encoding for the key enzyme in the polymerization of PHA monomers and other PHA-associated genes were identified from the genome. Phylogenetic analysis of the PhaC protein sequences has revealed that both PhaC1 and PhaC2 of Jeongeupia sp. USM3 are categorized as Class I PHA synthases with 56% similarity to each other. Both of the PHA synthase genes of this isolate were cloned and heterologously expressed in a PHA mutant strain Cupriavidus necator PHB-4. The ability of the transformants to accumulate PHA showed that both PhaC1 and PhaC2 were functional.
    Matched MeSH terms: Polyhydroxyalkanoates/biosynthesis*
  18. Genome Mining Reveals the Biosynthetic Pathways of Polyhydroxyalkanoate and Ectoines of the Halophilic Strain Salinivibrio proteolyticus M318 Isolated from Fermented Shrimp Paste
    Van Thuoc D, Loan TT, Trung TA, Van Quyen N, Tung QN, Tien PQ, et al.
    Mar Biotechnol (NY), 2020 Oct;22(5):651-660.
    PMID: 32827070 DOI: 10.1007/s10126-020-09986-z
    Salinivibrio proteolyticus M318, a halophilic bacterium isolated from fermented shrimp paste, is able to produce polyhydroxyalkanoate (PHA) from different carbon sources. In this study, we report the whole-genome sequence of strain M138, which comprises 2 separated chromosomes and 2 plasmids, and the complete genome contains 3,605,935 bp with an average GC content of 49.9%. The genome of strain M318 contains 3341 genes, 98 tRNA genes, and 28 rRNA genes. The 16S rRNA gene sequence and average nucleotide identity analysis associated with morphological and biochemical tests showed that this strain has high homology to the reference strain Salinivibrio proteolyticus DSM 8285. The genes encoding key enzymes for PHA and ectoine synthesis were identified from the bacterial genome. In addition, the TeaABC transporter responsible for ectoine uptake from the environment and the operon doeABXCD responsible for the degradation of ectoine were also detected. Strain M318 was able to produce poly(3-hydroxybutyrate) [P(3HB)] from different carbon sources such as glycerol, maltose, glucose, fructose, and starch. The ability to produce ectoines at different NaCl concentrations was investigated. High ectoine content of 26.2% of cell dry weight was obtained by this strain at 18% NaCl. This report provides genetic information regarding adaptive mechanisms of strain M318 to stress conditions, as well as new knowledge to facilitate the application of this strain as a bacterial cell factory for the production of PHA and ectoine.
    Matched MeSH terms: Polyhydroxyalkanoates/biosynthesis*
  19. Identification of regions affecting enzyme activity, substrate binding, dimer stabilization and polyhydroxyalkanoate (PHA) granule morphology in the PHA synthase of Aquitalea sp. USM4
    Lim H, Chuah JA, Chek MF, Tan HT, Hakoshima T, Sudesh K
    Int J Biol Macromol, 2021 Sep 01;186:414-423.
    PMID: 34246679 DOI: 10.1016/j.ijbiomac.2021.07.041
    Polyhydroxyalkanoates (PHAs) are biopolyesters synthesized by microorganisms as intracellular energy reservoirs under stressful environmental conditions. PHA synthase (PhaC) is the key enzyme responsible for PHA biosynthesis, but the importance of its N- and C-terminal ends still remains elusive. Six plasmid constructs expressing truncation variants of Aquitalea sp. USM4 PhaC (PhaC1As) were generated and heterologously expressed in Cupriavidus necator PHB-4. Removal of the first six residues at the N-terminus enabled the modulation of PHA composition without altering the PHA content in cells. Meanwhile, deletion of 13 amino acids from the C-terminus greatly affected the catalytic activity of PhaC1As, retaining only 1.1-7.4% of the total activity. Truncation(s) at the N- and/or C-terminus of PhaC1As gradually diminished the incorporation of comonomer units, and revealed that the N-terminal region is essential for PhaC1As dimerization whereas the C-terminal region is required for stabilization. Notably, transmission electron microscopy analysis showed that PhaC modification affected the morphology of intracellular PHA granules, which until now is only known to be regulated by phasins. This study provided substantial evidence and highlighted the significance of both the N- and C-termini of PhaC1As in regulating intracellular granule morphology, activity, substrate specificity, dimerization and stability of the synthase.
    Matched MeSH terms: Polyhydroxyalkanoates/metabolism*
  20. Fulltext Expression of Aeromonas caviae polyhydroxyalkanoate synthase gene in Burkholderia sp. USM (JCM15050) enables the biosynthesis of SCL-MCL PHA from palm oil products
    Chee JY, Lau NS, Samian MR, Tsuge T, Sudesh K
    J Appl Microbiol, 2012 Jan;112(1):45-54.
    PMID: 22054430 DOI: 10.1111/j.1365-2672.2011.05189.x
    Burkholderia sp. USM (JCM15050) isolated from oil-polluted wastewater is capable of utilizing palm oil products and glycerol to synthesize poly(3-hydroxybutyrate) [P(3HB)]. To confer the ability to produce polymer containing 3-hydroxyhexanoate (3HHx), plasmid (pBBREE32d13) harbouring the polyhydroxyalkanoate (PHA) synthase gene of Aeromonas caviae (phaC(Ac)) was transformed into this strain.
    Matched MeSH terms: Polyhydroxyalkanoates/biosynthesis*
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