Displaying publications 1 - 20 of 54 in total

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  1. Idris SN, Amelia TSM, Bhubalan K, Lazim AMM, Zakwan NAMA, Jamaluddin MI, et al.
    Environ Res, 2023 Aug 15;231(Pt 1):115988.
    PMID: 37105296 DOI: 10.1016/j.envres.2023.115988
    Plastics have become an integral part of human life. Single-use plastics (SUPs) are disposable plastics designed to be used once then promptly discarded or recycled. This SUPs range from packaging and takeaway containers to disposable razors and hotel toiletries. Synthetic plastics, which are made of non-renewable petroleum and natural gas resources, require decades to perpetually disintegrate in nature thus contribute to plastic pollution worldwide, especially in marine environments. In response to these problems, bioplastics or bio-based and biodegradable polymers from renewable sources has been considered as an alternative. Understanding the mechanisms behind the degradation of conventional SUPs and biodegradability of their greener counterpart, bioplastics, is crucial for appropriate material selection in the future. This review aims to provide insights into the degradation or disintegration of conventional single-use plastics and the biodegradability of the different types of greener-counterparts, bioplastics, their mechanisms, and conditions. This review highlights on the biodegradation in the environments including composting systems. Here, the various types of alternative biodegradable polymers, such as bacterially biosynthesised bioplastics, natural fibre-reinforced plastics, starch-, cellulose-, lignin-, and soy-based polymers were explored. Review of past literature revealed that although bioplastics are relatively eco-friendly, their natural compositions and properties are inconsistent. Furthermore, the global plastic market for biodegradable plastics remains relatively small and require further research and commercialization efforts, especially considering the urgency of plastic and microplastic pollution as currently critical global issue. Biodegradable plastics have potential to replace conventional plastics as they show biodegradation ability under real environments, and thus intensive research on the various biodegradable plastics is needed to inform stakeholders and policy makers on the appropriate response to the gradually emerging biodegradable plastics.
  2. Ganeson K, Alias AH, Murugaiyah V, Amirul AA, Ramakrishna S, Vigneswari S
    Pharmaceutics, 2023 Feb 23;15(3).
    PMID: 36986606 DOI: 10.3390/pharmaceutics15030744
    Cancer is the leading cause of death, acting as a global burden, severely impacting the patients' quality of life and affecting the world economy despite the expansion of cumulative advances in oncology. The current conventional therapies for cancer which involve long treatment duration and systemic exposure of drugs leads to premature degradation of drugs, a massive amount of pain, side effects, as well as the recurrence of the condition. There is also an urgent demand for personalized and precision-based medicine, especially after the recent pandemic, to avoid future delays in diagnosis or treatments for cancer patients as they are very essential in reducing the global mortality rate. Recently, microneedles which consist of a patch with tiny, micron-sized needles attached to it have been quite a sensation as an emerging technology for transdermal application to diagnose or treat various illnesses. The application of microneedles in cancer therapies is also being extensively studied as they offer a myriad of benefits, especially since microneedle patches offer a better treatment approach through self administration, painless treatment, and being an economically and environmentally friendly approach in comparison with other conventional methods. The painless gains from microneedles significantly improves the survival rate of cancer patients. The emergence of versatile and innovative transdermal drug delivery systems presents a prime breakthrough opportunity for safer and more effective therapies, which could meet the demands of cancer diagnosis and treatment through different application scenarios. This review highlights the types of microneedles, fabrication methods and materials, along with the recent advances and opportunities. In addition, this review also addresses the challenges and limitations of microneedles in cancer therapy with solutions through current studies and future works to facilitate the clinical translation of microneedles in cancer therapies.
  3. Jeremy Wong HS, Huong KH, Shafie NAH, Amirul AA
    J Biotechnol, 2021 Aug 20;337:71-79.
    PMID: 34233208 DOI: 10.1016/j.jbiotec.2021.07.001
    The sustainability in polyhydroxyalkanoates (PHA) production is drawing increasing attention as the effort to increase the economic feasibility for commercialization pursues. Oleic acid is widely preferred by bacteria but its employment for PHA production makes sustainability rather dubious. This study showed promising poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] content of 68 wt % by lipase genes-harbouring Cupriavidus malaysiensis USMAA2-4 transformant from palm olein and 1-pentanol. High oleic acid content and low oil saturation caused palm olein to outperform crude palm oil, crude palm kernel oil and soybean oil due to its preference for oleic acid shown by previous screening. The transformant showed 8-fold and 40-fold higher lipase activity compared to C. necator H16 and its wild-type respectively. The transformant was unaffected by Co2+ but the growth of C. necator H16 was inversely proportional to Co2+ concentration and the employment of 1-pentanol also ceased its growth and PHA accumulation. Although the inhibitory effect of Fe2+, Cu2+ and Zn2+ at high molarity on LipA decreased PHA content of C. malaysiensis USMAA2-4 transformant by 23-24 wt %, the lipase activity was restorable with high molarity of Ca2+, thus resulted in higher PHA content. The transformant enabled the employment of low-cost 1-pentanol as the precursor for cost-effective PHA production and its preference for palm olein contributed to higher sustainability.
  4. Vigneswari S, Noor MSM, Amelia TSM, Balakrishnan K, Adnan A, Bhubalan K, et al.
    Life (Basel), 2021 Aug 10;11(8).
    PMID: 34440551 DOI: 10.3390/life11080807
    Polyhydroxyalkanoates (PHA) are biodegradable polymers that are considered able to replace synthetic plastic because their biochemical characteristics are in some cases the same as other biodegradable polymers. However, due to the disadvantages of costly and non-renewable carbon sources, the production of PHA has been lower in the industrial sector against conventional plastics. At the same time, first-generation sugar-based cultivated feedstocks as substrates for PHA production threatens food security and considerably require other resources such as land and energy. Therefore, attempts have been made in pursuit of suitable sustainable and affordable sources of carbon to reduce production costs. Thus, in this review, we highlight utilising waste lignocellulosic feedstocks (LF) as a renewable and inexpensive carbon source to produce PHA. These waste feedstocks, second-generation plant lignocellulosic biomass, such as maize stoves, dedicated energy crops, rice straws, wood chips, are commonly available renewable biomass sources with a steady supply of about 150 billion tonnes per year of global yield. The generation of PHA from lignocellulose is still in its infancy, hence more screening of lignocellulosic materials and improvements in downstream processing and substrate pre-treatment are needed in the future to further advance the biopolymer sector.
  5. Vigneswari S, Gurusamy TP, Khairul WM, H P S AK, Ramakrishna S, Amirul AA
    Polymers (Basel), 2021 Jul 26;13(15).
    PMID: 34372060 DOI: 10.3390/polym13152454
    Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] is a bacterial derived biopolymer widely known for its unique physical and mechanical properties to be used in biomedical application. In this study, antimicrobial agent silver sulfadiazine (SSD) coat/collagen peptide coat-P(3HB-co-4HB) (SCCC) and SSD blend/collagen peptide coat-P(3HB-co-4HB) scaffolds (SBCC) were fabricated using a green salt leaching technique combined with freeze-drying. This was then followed by the incorporation of collagen peptides at various concentrations (2.5-12.5 wt.%) to P(3HB-co-4HB) using collagen-coating. As a result, two types of P(3HB-co-4HB) scaffolds were fabricated, including SCCC and SBCC scaffolds. The increasing concentrations of collagen peptides from 2.5 wt.% to 12.5 wt.% exhibited a decline in their porosity. The wettability and hydrophilicity increased as the concentration of collagen peptides in the scaffolds increased. In terms of the cytotoxic results, MTS assay demonstrated the L929 fibroblast scaffolds adhered well to the fabricated scaffolds. The 10 wt.% collagen peptides coated SCCC and SBCC scaffolds displayed highest cell proliferation rate. The antimicrobial analysis of the fabricated scaffolds exhibited 100% inhibition towards various pathogenic microorganisms. However, the SCCC scaffold exhibited 100% inhibition between 12 and 24 h, but the SBCC scaffolds with SSD impregnated in the scaffold had controlled release of the antimicrobial agent. Thus, this study will elucidate the surface interface-cell interactions of the SSD-P(3HB-co-4HB)-collagen peptide scaffolds and controlled release of SSD, antimicrobial agent.
  6. Huong KH, Sevakumaran V, Amirul AA
    Crit Rev Biotechnol, 2021 Jun;41(4):474-490.
    PMID: 33726581 DOI: 10.1080/07388551.2020.1869685
    Polyhydroxyalkanoate (PHA) is a biogenic polymer that has the potential to substitute synthetic plastic in numerous applications. This is due to its unique attribute of being a biodegradable and biocompatible thermoplastic, achievable through microbial fermentation from a broad utilizable range of renewable resources. Among all the PHAs discovered, poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] stands out as a next generation healthcare biomaterial for having high biopharmaceutical and medical value since it is highly compatible to mammalian tissue. This review provides a critical assessment and complete overview of the development and trend of P(3HB-co-4HB) research over the last few decades, highlighting aspects from the microbial strain discovery to metabolic engineering and bioprocess cultivation strategies. The article also outlines the relevance of P(3HB-co-4HB) as a material for high value-added products in numerous healthcare-related applications.
  7. Mohamed S, May Amelia TS, Abdullah Amirul AA, Abdul Wahid ME, Bhubalan K
    Biologicals, 2021 Jun;71:51-54.
    PMID: 33858743 DOI: 10.1016/j.biologicals.2021.03.002
    A natural biodegradable polymer, polyhydroxyalkanoate (PHA), was adjuvanted with a vaccine seed to observe the biomaterial's ability in enhancing an immune response in rats. The adjuvant potential of PHA was tested using the whole-killed Pasteurella multocida B:2 (PMB2) vaccine in Sprague Dawley (SD) rats to detect changes in serum immunoglobulin G (IgG) and immunoglobulin M (IgM) responses. A common PHA, poly(3-hydroxybutyrate) [P(3HB)], from Bacillus megaterium UMTKB-1 was constructed into microparticles using the solvent evaporation method. Twelve SD rats were divided into four treatment groups: 1) non-treatment as negative control, 2) P(3HB) adjuvant, 3) PMB2 vaccine, and 4) adjuvanted-P(3HB)/PMB2 vaccine groups, which were intramuscularly vaccinated twice. Immunoglobulins IgG and IgM levels were used as markers of the immune response induced by the adjuvanted-P(3HB)/PMB2 vaccine and analysed over an eight-week study period. The group vaccinated specifically with adjuvanted-P(3HB)/PMB2 vaccine had higher concentrations of immunoglobulins compared to other treatment groups, hence demonstrating the potential of the adjuvant to enhance immune response. Findings showed a need to delay the delivery of the second booster dose to determine the appropriate regime for the adjuvanted-P(3HB)/PMB2 vaccine.
  8. Oyekanmi AA, Kumar USU, H P S AK, Olaiya NG, Amirul AA, Rahman AA, et al.
    Polymers (Basel), 2021 May 20;13(10).
    PMID: 34065404 DOI: 10.3390/polym13101664
    Antimicrobial irradiated seaweed-neem biocomposite films were synthesized in this study. The storage functional properties of the films were investigated. Characterization of the prepared films was conducted using SEM, FT-IR, contact angle, and antimicrobial test. The macroscopic and microscopic including the analysis of the functional group and the gas chromatography-mass spectrometry test revealed the main active constituents present in the neem extract, which was used an essential component of the fabricated films. Neem leaves' extracts with 5% w/w concentration were incorporated into the matrix of seaweed biopolymer and the seaweed-neem bio-composite film were irradiated with different dosages of gamma radiation (0.5, 1, 1.5, and 2 kGy). The tensile, thermal, and the antimicrobial properties of the films were studied. The results revealed that the irradiated films exhibited improved functional properties compared to the control film at 1.5 kGy radiation dosage. The tensile strength, tensile modulus, and toughness exhibited by the films increased, while the elongation of the irradiated bio-composite film decreased compared to the control film. The morphology of the irradiated films demonstrated a smoother surface compared to the control and provided surface intermolecular interaction of the neem-seaweed matrix. The film indicated an optimum storage stability under ambient conditions and demonstrated no significant changes in the visual appearance. However, an increase in the moisture content was exhibited by the film, and the hydrophobic properties was retained until nine months of the storage period. The study of the films antimicrobial activities against Staphylococcus aureus (SA), and Bacillus subtilis (BS) indicated improved resistance to bacterial activities after the incorporation of neem leaves extract and gamma irradiation. The fabricated irradiated seaweed-neem bio-composite film could be used as an excellent sustainable packaging material due to its effective storage stability.
  9. Yahya EB, Amirul AA, H P S AK, Olaiya NG, Iqbal MO, Jummaat F, et al.
    Polymers (Basel), 2021 May 17;13(10).
    PMID: 34067569 DOI: 10.3390/polym13101612
    The global transplantation market size was valued at USD 8.4 billion in 2020 and is expected to grow at a compound annual growth rate of 11.5% over the forecast period. The increasing demand for tissue transplantation has inspired researchers to find alternative approaches for making artificial tissues and organs function. The unique physicochemical and biological properties of biopolymers and the attractive structural characteristics of aerogels such as extremely high porosity, ultra low-density, and high surface area make combining these materials of great interest in tissue scaffolding and regenerative medicine applications. Numerous biopolymer aerogel scaffolds have been used to regenerate skin, cartilage, bone, and even heart valves and blood vessels by growing desired cells together with the growth factor in tissue engineering scaffolds. This review focuses on the principle of tissue engineering and regenerative medicine and the role of biopolymer aerogel scaffolds in this field, going through the properties and the desirable characteristics of biopolymers and biopolymer tissue scaffolds in tissue engineering applications. The recent advances of using biopolymer aerogel scaffolds in the regeneration of skin, cartilage, bone, and heart valves are also discussed in the present review. Finally, we highlight the main challenges of biopolymer-based scaffolds and the prospects of using these materials in regenerative medicine.
  10. Vigneswari S, Amelia TSM, Hazwan MH, Mouriya GK, Bhubalan K, Amirul AA, et al.
    Antibiotics (Basel), 2021 Feb 24;10(3).
    PMID: 33668352 DOI: 10.3390/antibiotics10030229
    Nanobiotechnology has undoubtedly influenced major breakthroughs in medical sciences. Application of nanosized materials has made it possible for researchers to investigate a broad spectrum of treatments for diseases with minimally invasive procedures. Silver nanoparticles (AgNPs) have been a subject of investigation for numerous applications in agriculture, water treatment, biosensors, textiles, and the food industry as well as in the medical field, mainly due to their antimicrobial properties and nanoparticle nature. In general, AgNPs are known for their superior physical, chemical, and biological properties. The properties of AgNPs differ based on their methods of synthesis and to date, the biological method has been preferred because it is rapid, nontoxic, and can produce well-defined size and morphology under optimized conditions. Nevertheless, the common issue concerning biological or biobased production is its sustainability. Researchers have employed various strategies in addressing this shortcoming, such as recently testing agricultural biowastes such as fruit peels for the synthesis of AgNPs. The use of biowastes is definitely cost-effective and eco-friendly; moreover, it has been reported that the reduction process is simple and rapid with reasonably high yield. This review aims to address the developments in using fruit- and vegetable-based biowastes for biologically producing AgNPs to be applied as antimicrobial coatings in biomedical applications.
  11. Faisalina AF, Sonvico F, Colombo P, Amirul AA, Wahab HA, Majid MIA
    Nanomaterials (Basel), 2020 Oct 26;10(11).
    PMID: 33114572 DOI: 10.3390/nano10112123
    Polyhydroxyalkanoate (PHA) copolymers show a relatively higher in vivo degradation rate compared to other PHAs, thus, they receive a great deal of attention for a wide range of medical applications. Nanoparticles (NPs) loaded with poorly water-soluble anticancer drug docetaxel (DCX) were produced using poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB), copolymers biosynthesised from Cupriavidus malaysiensis USMAA1020 isolated from the Malaysian environment. Three copolymers with different molar proportions of 4-hydroxybutirate (4HB) were used: 16% (PHB16), 30% (PHB30) and 70% (PHB70) 4HB-containing P(3HB-co-4HB). Blank and DCX-loaded nanoparticles were then characterized for their size and size distribution, surface charge, encapsulation efficiency and drug release. Preformulation studies showed that an optimised formulation could be achieved through the emulsification/solvent evaporation method using PHB70 with the addition of 1.0% PVA, as stabilizer and 0.03% VitE-TPGS, as surfactant. DCX-loaded PHB70 nanoparticles (DCX-PHB70) gave the desired particle size distribution in terms of average particle size around 150 nm and narrow particle size distribution (polydispersity index (PDI) below 0.100). The encapsulation efficiency result showed that at 30% w/w drug-to-polymer ratio: DCX- PHB16 NPs were able to encapsulate up to 42% of DCX; DCX-PHB30 NPs encapsulated up to 46% of DCX and DCX-PHB70 NPs encapsulated up to 50% of DCX within the nanoparticle system. Approximately 60% of DCX was released from the DCX-PHB70 NPs within 7 days for 5%, 10% and 20% of drug-to-polymer ratio while for the 30% and 40% drug-to-polymer ratios, an almost complete drug release (98%) after 7 days of incubation was observed.
  12. Amelia TSM, Lau NS, Amirul AA, Bhubalan K
    Data Brief, 2020 Aug;31:105971.
    PMID: 32685631 DOI: 10.1016/j.dib.2020.105971
    Marine sponges are acknowledged as a bacterial hotspot and resource of novel natural products or genetic material with industrial or commercial potential. However, sponge-associated bacteria are difficult to be cultivated and the production of their desirable metabolites is inadequate in terms of rate and quantity, yet bioinformatics and metagenomics tools are steadily progressing. Bacterial diversity profiles of high-microbial-abundance wild tropical marine sponges Aaptos aaptos and Xestospongia muta were obtained by sample collection at Pulau Bidong and Pulau Redang islands, 16S rRNA amplicon sequencing on Illumina HiSeq2500 platform (250 bp paired-end) and metagenomics analysis using Ribosomal Database Project (RDP) classifier. Raw sequencing data in fastq format and relative abundance histograms of the dominant 10 species are available in the public repository Discover Mendeley Data (http://dx.doi.org/10.17632/zrcks5s8xp). Filtered sequencing data of operational taxonomic unit (OTU) with chimera removed is available in NCBI accession numbers from MT464469 to MT465036.
  13. Chai CJ, Amirul AA, Vigneswari S
    Data Brief, 2020 Feb;28:104777.
    PMID: 31871967 DOI: 10.1016/j.dib.2019.104777
    Electrospinning is a promising approach to fabricate desirable electropsun nanofibrous scaffold that could be applied in the medical fields. In this study, bacterial copolymer poly(3-hydroxybutyrate-co-68 mol% 4-hydroxybutyrate) [P(3HB-co-68mol% 4HB)] copolymer produced was fabricated into electrospun nanofibers using various combination of electrospinning parameters including the polymer solution, applied voltage and injection speed. The morphology of the fabricated scaffolds were observed using scanning electron microscope (SEM). The SEM images were analysed for the fibre diameter distribution of the scaffolds using Image Analyser. The results revealed that the 8 wt% of polymer solution, 25 kV/cm of the applied voltage and 1.5 mL/h of the injection speed was the most suitable combination. This electrospinning parameters combination fabricated nanofibrous P(3HB-co-4HB) scaffold with smooth, beadles and uniform nanofibers with small fibre diameter distribution.
  14. Vigneswari S, Chai JM, Kamarudin KH, Amirul AA, Focarete ML, Ramakrishna S
    Front Bioeng Biotechnol, 2020;8:567693.
    PMID: 33195129 DOI: 10.3389/fbioe.2020.567693
    Biomaterial scaffolds play crucial role to promote cell proliferation and foster the regeneration of new tissues. The progress in material science has paved the way for the generation of ingenious biomaterials. However, these biomaterials require further optimization to be effectively used in existing clinical treatments. It is crucial to develop biomaterials which mimics structure that can be actively involved in delivering signals to cells for the formation of the regenerated tissue. In this research we nanoengineered a functional scaffold to support the proliferation of myoblast cells. Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] copolymer is chosen as scaffold material owing to its desirable mechanical and physical properties combined with good biocompatibility, thus eliciting appropriate host tissue responses. In this study P(3HB-co-4HB) copolymer was biosynthesized using Cupriavidus malaysiensis USMAA1020 transformant harboring additional PHA synthase gene, and the viability of a novel P(3HB-co-4HB) electrospun nanofiber scaffold, surface functionalized with RGD peptides, was explored. In order to immobilize RGD peptides molecules onto the P(3HB-co-4HB) nanofibers surface, an aminolysis reaction was performed. The nanoengineered scaffolds were characterized using SEM, organic elemental analysis (CHN analysis), FTIR, surface wettability and their in vitro degradation behavior was evaluated. The cell culture study using H9c2 myoblast cells was conducted to assess the in vitro cellular response of the engineered scaffold. Our results demonstrated that nano-P(3HB-co-4HB)-RGD scaffold possessed an average fiber diameter distribution between 200 and 300 nm, closely biomimicking, from a morphological point of view, the structural ECM components, thus acting as potential ECM analogs. This study indicates that the surface conjugation of biomimetic RGD peptide to the nano-P(3HB-co-4HB) fibers increased the surface wettability (15 ± 2°) and enhanced H9c2 myoblast cells attachment and proliferation. In summary, the study reveals that nano-P(3HB-co-4HB)-RGD scaffold can be considered a promising candidate to be further explored as cardiac construct for building cardiac construct.
  15. Sam KK, Lau NS, Furusawa G, Amirul AA
    Microbiol Resour Announc, 2019 Nov 14;8(46).
    PMID: 31727719 DOI: 10.1128/MRA.01248-19
    Pararhodobacter-like strain CCB-MM2 is a halophilic alphaproteobacterium isolated from estuarine sediment collected from Matang Mangrove Forest in Malaysia. Here, we present the draft genome sequence of CCB-MM2 and provide insights into its physiological roles and metabolic potential.
  16. Ali MS, Isa NM, Abedelrhman FM, Alyas TB, Mohammed SE, Ahmed AE, et al.
    BMC Microbiol, 2019 06 11;19(1):126.
    PMID: 31185900 DOI: 10.1186/s12866-019-1470-2
    BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is known as a leading cause of morbidity and mortality. Investigation of the MRSA's virulence and resistance mechanisms is a continuing concern toward controlling such burdens through using high throughput whole Genome Sequencing (WGS) and molecular diagnostic assays. The objective of the present study is to perform whole-genome sequencing of MRSA isolated from Sudan using Illumina Next Generation Sequencing (NGS) platform.

    RESULTS: The genome of MRSA strain SO-1977 consists of 2,827,644 bp with 32.8% G + C, 59 RNAs and 2629 predicted coding sequences (CDSs). The genome has 26 systems, one of which is the major class in the disease virulence and defence. A total of 83 genes were annotated to virulence disease and defence category some of these genes coding as functional proteins. Based on genome analysis, it is speculated that the SO-1977 strain has resistant genes to Teicoplanin, Fluoroquinolones, Quinolone, Cephamycins, Tetracycline, Acriflavin and Carbapenems. The results revealed that the SO-1977, strain isolated from Sudan has a wide range of antibiotic resistance compared to related strains.

    CONCLUSION: The study reports for the first time the whole genome sequence of Sudan MRSA isolates. The release of the genome sequence of the strain SO-1977 will avail MRSA in public databases for further investigations on the evolution of resistant mechanism and dissemination of the -resistant genes of MRSA.

  17. 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.
  18. Norhafini H, Huong KH, Amirul AA
    Int J Biol Macromol, 2019 Mar 15;125:1024-1032.
    PMID: 30557643 DOI: 10.1016/j.ijbiomac.2018.12.121
    P(3HB-co-4HB) with a high 4HB monomer composition was previously successfully produced using the transformant Cupriavidus malaysiensis USMAA1020 containing an additional copy of the PHA synthase gene. In this study, high PHA density fed-batch cultivation strategies were developed for such 4HB-rich P(3HB-co-4HB). The pulse, constant and mixed feeding strategies resulted in high PHA accumulation, with a PHA content of 74-92 wt% and 4HB monomer composition of 92-99 mol%. The pulse-feed of carbon and nitrogen resulted in higher PHA concentration (30.7 g/L) than carbon alone (22.3 g/L), suggesting that a trace amount of nitrogen is essential to support cell density for PHA accumulation. Constant feeding was found to be a more feasible strategy than mixed feeding, since the latter caused a drastic fluctuation in the C/N ratio, as evidenced by higher biomass formation indicating more carbon flux towards the competitive TCA pathway. A two-times carbon and nitrogen pulse feeding was the most optimal strategy achieving 92 wt% accommodation of the total biomass, with the highest PHA concentration (46 g/L) and yield (Yp/x) of 11.5 g/g. The strategy has kept the C/N at optimal ratio during the active PHA-producing phase. This is the first report of the production of high PHA density for 4HB-rich P(3HB-co-4HB).
  19. 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.
  20. Huong KH, Elina KAR, Amirul AA
    Int J Biol Macromol, 2018 Sep;116:217-223.
    PMID: 29723627 DOI: 10.1016/j.ijbiomac.2018.04.148
    Long carbon chain alkanediols are used in the production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)], however these substrates possess high toxicity towards bacterial cells. This study demonstrated the effective utilisation of a long carbon chain alkanediol, namely 1,8-octanediol, to enhance the yield and production of a copolymer with a high molecular weight of over 1000 kDa, which is desirable for novel applications in medical and biopharmaceuticals. The increased PHA content (47-61 wt%) and concentration (1.7-4.5 g/L) was achieved by additional feeding of a combination of C4 substrates at C/N 10, with 1,8-octanediol + γ-butyrolactone producing P(3HB-co-22 mol% 4HB) with a high molecular weight (1060 kDa) and elongation at break of 970%. The DO-stat feeding strategy of C/N 10 has shown an increment of PHA concentration for both carbon combination, 0.45-4.27 g/L and 0.32-3.36 g/L for 1,8-octanediol + sodium 4-hydroxybutyrate (4HB-Na) and 1,8-octanediol + γ-butyrolactone, but with a slight reduction on molecular weight and mechanical strength. Nonetheless, further study revealed that a nitrogen-absence feeding strategy could retain the high molecular weight and elongation at break of the copolymer, and simultaneously improving the overall P(3HB-co-4HB) production.
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