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Biomimetic growth of bone-like apatite via simulated body fluid on hydroxyethyl cellulose/polyvinyl alcohol electrospun nanofibers

Chahal S, Chalal S, Fathima SJ, Yusoff MB

Biomed Mater Eng, 2014;24(1):799-806.
PMID: 24211966 DOI: 10.3233/BME-130871

In this study, randomly oriented hydroxyethyl cellulose/polyvinyl alcohol (HEC/PVA) nanofibers were fabricated by electrospinning. The blend solutions of HEC/PVA with different weight ratio of HEC to PVA were prepared using water as solvent to fabricate nanofibers. These nanofibrous scaffolds were coated with bone-like apatite by immersing into 10x simulated body fluid (SBF) for different time periods. The morphology and structure of the nanofibers were characterized by SEM, FTIR and DSC. FESEM-EDS and FTIR analysis were used to confirm the deposition of apatite on the surface of nanofibers. The results of this study suggest that this apatite coated nanofibrous scaffolds could be a suitable biomaterial for bone tissue engineering.

Matched MeSH terms: Calcium Phosphates/chemistry
Influence of Buffer Composition and Calcium Chloride on GdnHCl Denaturation of Bacillus licheniformis α-Amylase

Kandandapani S, Tan CY, Shuib AS, Tayyab S

Protein Pept Lett, 2016;23(6):537-43.
PMID: 26936029

The influence of buffer composition on the conformational stability of native and calciumdepleted Bacillus licheniformis α-amylase (BLA) was investigated against guanidine hydrochloride (GdnHCl) denaturation using circular dichroism, fluorescence and UV-difference spectroscopy. Differential effect of buffer composition on GdnHCl denaturation of BLA was evident from the magnitude of these spectral signals, which followed the order: sodium phosphate > Tris-HCl > HEPES > MOPS. These effects became more pronounced with calcium-depleted BLA. Sephacryl S-200 gel chromatographic results showed significant BLA aggregation in the presence of 6 M GdnHCl.

Matched MeSH terms: Phosphates/chemistry*
Reduction of Mo(VI) by the bacterium Serratia sp. strain DRY5

Rahman MF, Shukor MY, Suhaili Z, Mustafa S, Shamaan NA, Syed MA

J Environ Biol, 2009 Jan;30(1):65-72.
PMID: 20112865

The need to isolate efficient heavy metal reducers for cost effective bioremediation strategy have resulted in the isolation of a potent molybdenum-reducing bacterium. The isolate was tentatively identified as Serratia sp. strain DRY5 based on the Biolog GN carbon utilization profiles and partial 16S rDNA molecular phylogeny. Strain DRY5 produced 2.3 times the amount of Mo-blue than S. marcescens strain Dr.Y6, 23 times more than E. coli K12 and 7 times more than E. cloacae strain 48. Strain DRY5 required 37 degrees C and pH 7.0 for optimum molybdenum reduction. Carbon sources such as sucrose, maltose, glucose and glycerol, supported cellular growth and molybdate reduction after 24 hr of static incubation. The most optimum carbon source that supported reduction was sucrose at 1.0% (w/v). Ammonium sulphate, ammonium chloride, glutamic acid, cysteine, and valine supported growth and molybdate reduction with ammonium sulphate as the optimum nitrogen source at 0. 2% (w/v). Molybdate reduction was optimally supported by 30 mM molybdate. The optimum concentration of phosphate for molybdate reduction was 5 mM when molybdate concentration was fixed at 30 mM and molybdate reduction was totally inhibited at 100 mM phosphate. Mo-blue produced by this strain shows a unique characteristic absorption profile with a maximum peak at 865 nm and a shoulder at 700 nm, Dialysis tubing experiment showed that 95.42% of Mo-blue was found in the dialysis tubing suggesting that the molybdate reduction seen in this bacterium was catalyzed by enzyme(s). The characteristics of isolate DRY5 suggest that it would be useful in the bioremediation ofmolybdenum-containing waste.

Matched MeSH terms: Phosphates/chemistry
Fulltext Kinetics of molybdenum reduction to molybdenum blue by Bacillus sp. strain A.rzi

Othman AR, Bakar NA, Halmi MI, Johari WL, Ahmad SA, Jirangon H, et al.

Biomed Res Int, 2013;2013:371058.
PMID: 24369531 DOI: 10.1155/2013/371058

Molybdenum is very toxic to agricultural animals. Mo-reducing bacterium can be used to immobilize soluble molybdenum to insoluble forms, reducing its toxicity in the process. In this work the isolation of a novel molybdate-reducing Gram positive bacterium tentatively identified as Bacillus sp. strain A.rzi from a metal-contaminated soil is reported. The cellular reduction of molybdate to molybdenum blue occurred optimally at 4 mM phosphate, using 1% (w/v) glucose, 50 mM molybdate, between 28 and 30 °C and at pH 7.3. The spectrum of the Mo-blue product showed a maximum peak at 865 nm and a shoulder at 700 nm. Inhibitors of bacterial electron transport system (ETS) such as rotenone, sodium azide, antimycin A, and potassium cyanide could not inhibit the molybdenum-reducing activity. At 0.1 mM, mercury, copper, cadmium, arsenic, lead, chromium, cobalt, and zinc showed strong inhibition on molybdate reduction by crude enzyme. The best model that fitted the experimental data well was Luong followed by Haldane and Monod. The calculated value for Luong's constants p max, K(s), S(m), and n was 5.88 μmole Mo-blue hr(-1), 70.36 mM, 108.22 mM, and 0.74, respectively. The characteristics of this bacterium make it an ideal tool for bioremediation of molybdenum pollution.

Matched MeSH terms: Phosphates/chemistry
Tropical stormwater nutrient degradation using nano-TiO2 in photocatalytic reactor detention pond

Rad S, Shamsudin S, Taha MR, Shahid S

Water Sci Technol, 2016;73(2):405-13.
PMID: 26819397 DOI: 10.2166/wst.2015.465

The photo-degradation of nutrients in stormwater in photocatalytic reactor wet detention pond using nano titanium dioxide (TiO2) in concrete was investigated in a scale model as a new stormwater treatment method. Degradation of phosphate and nitrate in the presence of nano-TiO2 under natural ultra violet (UV) from tropical sunlight was monitored for 3 weeks compared with normal ponds. Two types of cement, including ordinary Portland and white cement mixed with TiO2 nano powder, were used as a thin cover to surround the body of the pond. Experiments with and without the catalyst were carried out for comparison and control. Average Anatase diameter of 25 nm and Rutile 100 nm nano particles were applied at three different mixtures of 3, 10 and 30% weight. The amounts of algae available orthophosphate and nitrate, which cause eutrophication in the ponds, were measured during the tests. Results revealed that the utilization of 3% up to 30% weight nano-TiO2 can improve stormwater outflow quality by up to 25% after 48 h and 57% after 3 weeks compared with the control sample in normal conditions with average nutrient (phosphate and nitrate) removal of 4% after 48 h and 10% after 3 weeks.

Matched MeSH terms: Phosphates/chemistry*
Ion-Doped Silicate Bioceramic Coating of Ti-Based Implant

Mohammadi H, Sepantafar M

Iran Biomed J, 2016 Sep;20(4):189-200.
PMID: 26979401

Titanium and its alloy are known as important load-bearing biomaterials. The major drawbacks of these metals are fibrous formation and low corrosion rate after implantation. The surface modification of biomedical implants through various methods such as plasma spray improves their osseointegration and clinical lifetime. Different materials have been already used as coatings on biomedical implant, including calcium phosphates and bioglass. However, these materials have been reported to have limited clinical success. The excellent bioactivity of calcium silicate (Ca-Si) has been also regarded as coating material. However, their high degradation rate and low mechanical strength limit their further coating application. Trace element modification of (Ca-Si) bioceramics is a promising method, which improves their mechanical strength and chemical stability. In this review, the potential of trace element-modified silicate coatings on better bone formation of titanium implant is investigated.

Matched MeSH terms: Calcium Phosphates/chemistry
Role of phosphate solubilizing bacteria on rock phosphate solubility and growth of aerobic rice

Panhwar QA, Radziah O, Zaharah AR, Sariah M, Razi IM

J Environ Biol, 2011 Sep;32(5):607-12.
PMID: 22319876

Use of phosphate-solubilizing bacteria (PSB) as inoculants has concurrently increased phosphorous uptake in plants and improved yields in several crop species. The ability of PSB to improve growth of aerobic rice (Oryza sativa L.) through enhanced phosphorus (P) uptake from Christmas island rock phosphate (RP) was studied in glasshouse experiments. Two isolated PSB strains; Bacillus spp. PSB9 and PSB16, were evaluated with RP treatments at 0, 30 and 60 kg ha(-1). Surface sterilized seeds of aerobic rice were planted in plastic pots containing 3 kg soil and the effect of treatments incorporated at planting were observed over 60 days of growth. The isolated PSB strains (PSB9 and PSB16) solubilized significantly high amounts of P (20.05-24.08 mg kg(-1)) compared to non-inoculated (19-23.10 mg kg(-1)) treatments. Significantly higher P solubilization (24.08 mg kg(-1)) and plant P uptake (5.31 mg plant(-1)) was observed with the PSB16 strain at the highest P level of 60 kg ha(-1). The higher amounts of soluble P in the soil solution increased P uptake in plants and resulted in higher plant biomass (21.48 g plant(-1)). PSB strains also increased plant height (80 cm) and improved root morphology in aerobic rice. The results showed that inoculation of aerobic rice with PSB improved phosphate solubilizing activity of incorporated RP.

Matched MeSH terms: Phosphates/chemistry*
Mechanical and cytotoxicity properties of hybrid ceramics filled polyamide 12 filament feedstock for craniofacial bone reconstruction via fused deposition modelling

Abdullah AM, Rahim TNAT, Hamad WNFW, Mohamad D, Akil HM, Rajion ZA

Dent Mater, 2018 11;34(11):e309-e316.
PMID: 30268678 DOI: 10.1016/j.dental.2018.09.006

OBJECTIVE: To compare the mechanical and biological properties of newly developed hybrid ceramics filled and unfilled polyamide 12 (PA 12) for craniofacial reconstruction via a fused deposition modelling (FDM) framework.
METHODS: 15wt% of zirconia (ZrO2) as well as 30, 35, and 40wt% of beta-tricalcium phosphate (β-TCP) were compounded with PA 12, followed by the fabrication of filament feedstocks using a single screw extruder. The fabricated filament feedstocks were used to print the impact specimens. The melt flow rate, tensile properties of fabricated filament feedstocks, and 3D printed impact properties of the specimens were assessed using melt flow indexer, universal testing machine, and Izod pendulum tester, respectively. The microstructure of selected filament feedstocks and broken impact specimens were analysed using a field emission scanning electron microscope and universal testing machine. Human periodontal ligament fibroblast cells (HPdLF) were used to evaluate the cytotoxicity of the materials by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid) (MTT) assay.
RESULTS: Hybrid ceramics filled PA 12 indicated sufficient flowability for FDM 3D printing. The tensile strength of hybrid ceramics filled PA 12 filament feedstocks slightly reduced as compared to unfilled PA 12. However, the tensile modulus and impact strength of hybrid ceramics filled PA 12 increased by 8%-31% and 98%-181%, respectively. A significant increase was also detected in the cell viability of the developed composites at concentrations of 12.5, 25, 50 and 100mg/ml.
SIGNIFICANCE: The newly developed hybrid ceramics filled PA 12 filament feedstock with improved properties is suitable for an FDM-based 3D printer, which enables the creation of patient-specific craniofacial implant at a lower cost to serve low-income patients.

Matched MeSH terms: Calcium Phosphates/chemistry
Enhancement of thermal, mechanical and physical properties of polyamide 12 composites via hybridization of ceramics for bone replacement

Abdullah AM, Mohamad D, Rahim TNAT, Akil HM, Rajion ZA

Mater Sci Eng C Mater Biol Appl, 2019 Jun;99:719-725.
PMID: 30889745 DOI: 10.1016/j.msec.2019.02.007

This study reports the influence of ZrO2/β-TCP hybridization on the thermal, mechanical, and physical properties of polyamide 12 composites to be suited for bone replacement. Amount of 15 wt% of nano-ZrO2 along with 5,10,15,20 and 25 wt% of micro-β-TCP was compounded with polyamide 12 via a twin-screw extruder. The hybrid ZrO2/β-TCP filled polyamide 12 exhibited higher thermal, mechanical and physical properties in comparison to unfilled polyamide 12 at certain filler loading; which is attributed to the homogenous dispersion of ZrO2/β-TCP fillers particle in polyamide 12 matrix. The hybrid ZrO2/β-TCP filled PA 12 demonstrated an increment of tensile strength by up to 1%, tensile modulus of 38%, flexural strength of 15%, flexural modulus of 45%, and surface roughness value of 93%, as compared to unfilled PA 12. With enhanced thermal, mechanical and physical properties, the newly developed hybrid ZrO2/β-TCP filled PA 12 could be potentially utilized for bone replacement.

Matched MeSH terms: Calcium Phosphates/chemistry
Development of a bone substitute material based on alpha-tricalcium phosphate scaffold coated with carbonate apatite/poly-epsilon-caprolactone

Bang LT, Ramesh S, Purbolaksono J, Long BD, Chandran H, Ramesh S, et al.

Biomed Mater, 2015 Aug;10(4):045011.
PMID: 26225725 DOI: 10.1088/1748-6041/10/4/045011

Interconnected porous tricalcium phosphate ceramics are considered to be potential bone substitutes. However, insufficient mechanical properties when using tricalcium phosphate powders remain a challenge. To mitigate these issues, we have developed a new approach to produce an interconnected alpha-tricalcium phosphate (α-TCP) scaffold and to perform surface modification on the scaffold with a composite layer, which consists of hybrid carbonate apatite / poly-epsilon-caprolactone (CO3Ap/PCL) with enhanced mechanical properties and biological performance. Different CO3Ap combinations were tested to evaluate the optimal mechanical strength and in vitro cell response of the scaffold. The α-TCP scaffold coated with CO3Ap/PCL maintained a fully interconnected structure with a porosity of 80% to 86% and achieved an improved compressive strength mimicking that of cancellous bone. The addition of CO3Ap coupled with the fully interconnected microstructure of the α-TCP scaffolds coated with CO3Ap/PCL increased cell attachment, accelerated proliferation and resulted in greater alkaline phosphatase (ALP) activity. Hence, our bone substitute exhibited promising potential for applications in cancellous bone-type replacement.

Matched MeSH terms: Calcium Phosphates/chemistry*
Macro-nutrients recovery from liquid waste as a sustainable resource for production of recovered mineral fertilizer: Uncovering alternative options to sustain global food security cost-effectively

Sniatala B, Kurniawan TA, Sobotka D, Makinia J, Othman MHD

Sci Total Environ, 2023 Jan 15;856(Pt 2):159283.
PMID: 36208738 DOI: 10.1016/j.scitotenv.2022.159283

Global food security, which has emerged as one of the sustainability challenges, impacts every country. As food cannot be generated without involving nutrients, research has intensified recently to recover unused nutrients from waste streams. As a finite resource, phosphorus (P) is largely wasted. This work critically reviews the technical applicability of various water technologies to recover macro-nutrients such as P, N, and K from wastewater. Struvite precipitation, adsorption, ion exchange, and membrane filtration are applied for nutrient recovery. Technological strengths and drawbacks in their applications are evaluated and compared. Their operational conditions such as pH, dose required, initial nutrient concentration, and treatment performance are presented. Cost-effectiveness of the technologies for P or N recovery is also elaborated. It is evident from a literature survey of 310 published studies (1985-2022) that no single technique can effectively and universally recover target macro-nutrients from liquid waste. Struvite precipitation is commonly used to recover over 95 % of P from sludge digestate with its concentration ranging from 200 to 4000 mg/L. The recovered precipitate can be reused as a fertilizer due to its high content of P and N. Phosphate removal of higher than 80 % can be achieved by struvite precipitation when the molar ratio of Mg2+/PO43- ranges between 1.1 and 1.3. The applications of artificial intelligence (AI) to collect data on critical parameters control optimization, improve treatment effectiveness, and facilitate water utilities to upscale water treatment plants. Such infrastructure in the plants could enable the recovered materials to be reused to sustain food security. As nutrient recovery is crucial in wastewater treatment, water treatment plant operators need to consider (1) the costs of nutrient recovery techniques; (2) their applicability; (3) their benefits and implications. It is essential to note that the treatment cost of P and/or N-laden wastewater depends on the process applied and local conditions.

Matched MeSH terms: Phosphates/chemistry
Fulltext In vivo efficacy of tobramycin-loaded synthetic calcium phosphate beads in a rabbit model of staphylococcal osteomyelitis

Lulu GA, Karunanidhi A, Mohamad Yusof L, Abba Y, Mohd Fauzi F, Othman F

Ann Clin Microbiol Antimicrob, 2018 Dec 28;17(1):46.
PMID: 30593272 DOI: 10.1186/s12941-018-0296-3

BACKGROUND: Osteomyelitis is an acute or chronic inflammatory process of the bone following infection with pyogenic organisms like Staphylococcus aureus. Tobramycin (TOB) is a promising aminoglycoside antibiotic used to treat various bacterial infections, including S. aureus. The aim of this study was to investigate the efficacy of tobramycin-loaded calcium phosphate beads (CPB) in a rabbit osteomyelitis model.
METHODS: Tobramycin (30 mg/mL) was incorporated into CPB by dipping method and the efficacy of TOB-loaded CPB was studied in a rabbit osteomyelitis model. For juxtaposition, CPB with and without TOB were prepared. Twenty-five New Zealand white rabbits were grouped (n = 5) as sham (group 1), TOB-loaded CPB without S. aureus (group 2), S. aureus only (group 3), S. aureus + CPB (group 4), and S. aureus + TOB-loaded CPB (group 5). Groups infected with S. aureus followed by CPB implantation were immediately subjected to surgery at the mid-shaft of the tibia. After 28 days post-surgery, all rabbits were euthanized and the presence or absence of chronic osteomyelitis and the extent of architectural destruction of the bone were assessed by radiology, bacteriology and histological studies.
RESULTS: Tobramycin-loaded CPB group potentially inhibited the growth of S. aureus causing 3.2 to 3.4 log10 reductions in CFU/g of bone tissue compared to the controls. Untreated groups infected with S. aureus showed signs of chronic osteomyelitis with abundant bacterial growth and alterations in bone architecture. The sham group and TOB-loaded CPB group showed no evidence of bacterial growth.
CONCLUSIONS: TOB-incorporated into CPB for local bone administration was proven to be more successful in increasing the efficacy of TOB in this rabbit osteomyelitis model and hence could represent a good alternative to other formulations used in the treatment of osteomyelitis.

Matched MeSH terms: Calcium Phosphates/chemistry*
Fulltext Reduction of saltiness and acrylamide levels in palm sugar-like flavouring through buffer modification and the addition of calcium chloride

Tan PY, Tan CP, Abas F, Ho CW, Mustapha WA

Molecules, 2013 Jun 10;18(6):6792-803.
PMID: 23752466 DOI: 10.3390/molecules18066792

Palm sugar-like flavouring (PSLF) is a type of flavour product that is formed by heating amino acids and sugar under specific heating conditions. Unfortunately, PSLF has a salty taste and contains high amounts of acrylamide. Hence, the objective of this research was to reduce saltiness and acrylamide without negatively affecting the aroma properties of PSLF. A decrease in the sodium phosphate (NaHPO₄) buffer concentration from 0.20 to 0.02 M was found to reduce sodium to approximately 15% of the level found in original PSLF. A further decrease (~25%) in the sodium content was achieved by removing monobasic sodium phosphate (NaH₂PO₄) from the buffer system. Meanwhile, the addition of CaCl₂ at 20-40 mg/L reduced the acrylamide content in PSLF by as much as 58%. A CaCl₂ concentration of 20 mg/mL was most favourable as it most efficiently suppressed acrylamide formation while providing an acceptably high flavour yield in PSLF. In view of the high acrylamide content in PSLF, additional work is necessary to further reduce the amount of acrylamide by controlling the asparagine concentration in the precursor mixture.

Matched MeSH terms: Phosphates/chemistry
3D-printed biphasic calcium phosphate scaffolds coated with an oxygen generating system for enhancing engineered tissue survival

Touri M, Moztarzadeh F, Osman NAA, Dehghan MM, Mozafari M

Mater Sci Eng C Mater Biol Appl, 2018 Mar 01;84:236-242.
PMID: 29519434 DOI: 10.1016/j.msec.2017.11.037

Tissue engineering scaffolds with oxygen generating elements have shown to be able to increase the level of oxygen and cell survivability in specific conditions. In this study, biphasic calcium phosphate (BCP) scaffolds with the composition of 60% hydroxyapatite (HA) and 40% beta-tricalcium phosphate (β-TCP), which have shown a great potential for bone tissue engineering applications, were fabricated by a direct-write assembly (robocasting) technique. Then, the three-dimensional (3D)-printed scaffolds were coated with different ratios of an oxygen releasing agent, calcium peroxide (CPO), which encapsulated within a polycaprolactone (PCL) matrix through dip-coating, and used for in situ production of oxygen in the implanted sites. The structure, composition and morphology of the prepared scaffolds were characterized by different techniques. The oxygen release kinetics and biological investigations of the scaffolds were also studied in vitro. The results showed that oxygen release behaviour was sustained and dependant on the concentration of CPO encapsulated in the PCL coating matrix. It was also demonstrated that the coated scaffolds, having 3% CPO in the coating system, could provide a great potential for promoting bone ingrowth with improving osteoblast cells viability and proliferation under hypoxic conditions. The findings indicated that the prepared scaffolds could play a significant role in engineering of large bone tissue implants with limitations in oxygen diffusion.

Matched MeSH terms: Calcium Phosphates/chemistry*
Synthesis and characterization of magnesium doped biphasic calcium phosphate

Toibah AR, Sopyan I, Mel M

Med J Malaysia, 2008 Jul;63 Suppl A:83-4.
PMID: 19024995

The incorporation of magnesium ions into the calcium phosphate structure is of great interest for the development of artificial bone implants. This paper investigates the preparation of magnesium-doped biphasic calcium phosphate (Mg-BCP) via sol gel method at various concentrations of added Mg. The effect of calcinations temperature (ranging from 500 degrees C to 900 degrees C) and concentrations of Mg incorporated into BCP has been studied by the aid of XRD, TGA and infrared spectroscopy (IR) in transmittance mode analysis. The study indicated that the powder was pure BCP and Mg-BCP with 100% purity and high crystallinity. The results also indicated that beta-tricalcium phosphate (beta-TCP) phase can be observed when the powder was calcined at 800 degrees C and above.

Matched MeSH terms: Calcium Phosphates/chemistry*
Nutrient removal in a pilot and full scale constructed wetland, Putrajaya city, Malaysia

Sim CH, Yusoff MK, Shutes B, Ho SC, Mansor M

J Environ Manage, 2008 Jul;88(2):307-17.
PMID: 17467147

Putrajaya Wetlands in Malaysia, a 200ha constructed wetland system consisting of 24 cells, was created in 1997-1998 to treat surface runoff caused by development and agricultural activities from an upstream catchment before entering Putrajaya Lake (400ha). It was designed for stormwater treatment, flood control and amenity use. The water quality improvement performance of a section of the wetland cells is described. The nutrient removal performance was 82.11% for total nitrogen, 70.73% for nitrate-nitrogen and 84.32% for phosphate, respectively, along six wetland cells from Upper North UN6 to UN1 from April to December 2004. Nutrient removal in pilot scale tank systems, simulating a constructed wetland and planted with examples of common species at Putrajaya, the Common Reed Phragmites karka and Tube Sedge Lepironia articulata, and the capacity of these species to retain nutrients in above and below-ground plant biomass and substrate is reported. The uptake of nutrients by the Common Reed and Tube Sedge from the pilot tank system was 42.1% TKN; 28.9% P and 17.4% TKN; 26.1% P, respectively. The nutrient uptake efficiency of the Common Reed was higher in above-ground than in below-ground tissue. The results have implications for plant species selection in the design of constructed wetlands in Malaysia and for optimizing the performance of these systems.

Matched MeSH terms: Phosphates/chemistry
Capillary electrophoresis with capacitively coupled contactless conductivity detection for the determination of cis/trans isomers of octadec-9-enoic acid and other long chain fatty acids

Wong YF, Saad B, Makahleh A

J Chromatogr A, 2013 May 17;1290:82-90.
PMID: 23578483 DOI: 10.1016/j.chroma.2013.03.014

A capillary electrophoresis (CE)-capacitively coupled contactless conductivity detection (C(4)D) method for the simultaneous separation of eleven underivatized fatty acids (FAs), namely, lauric, myristic, tridecanoic (internal standard), pentadecanoic, palmitic, stearic, oleic, elaidic, linoleic, linolenic and arachidic acids is described. The separation was carried out in normal polarity mode at 20 °C, 30 kV and using hydrodynamic injection (50 mbar for 1 s). The separation was achieved in a bare fused-silica capillary (70 cm × 75 μm i.d.) using a background electrolyte of methyl-β-cyclodextrin (~6 mM) and heptakis-(2,3,6-tri-O-methyl)-β-cyclodextrin (~8 mM) dissolved in a mixture of Na2HPO4/KH2PO4 (5 mM, pH 7.4):ACN:MeOH:n-octanol (3:4:2.5:0.5, v/v/v/v). C(4)D parameters were set at fixed amplitude of 100 V and frequency of 1000 kHz. The developed method was validated. Calibration curves of the ten FAs were well correlated (r(2)>0.99) within the range of 5-250 μg mL(-1) for lauric acid, and 3-250 μg mL(-1) for the other FAs. The method was simple and sensitive with detection limits (S/N=3) of 0.9-1.9 μg mL(-1) and good relative standard deviations of intra- and inter-day for migration times and peak areas (≤9.7%) were achieved. The method was applied to the determination of FAs in margarine samples. The proposed method offers distinct advantages over the GC and HPLC methods, especially in terms of simplicity (without derivatization) and sensitivity.

Matched MeSH terms: Phosphates/chemistry
Production of γ-cyclodextrin by Bacillus cereus cyclodextrin glycosyltransferase using extractive bioconversion in polymer-salt aqueous two-phase system

Lin YK, Show PL, Yap YJ, Ariff AB, Mohammad Annuar MS, Lai OM, et al.

J Biosci Bioeng, 2016 Jun;121(6):692-696.
PMID: 26702953 DOI: 10.1016/j.jbiosc.2015.11.001

Aqueous two-phase system (ATPS) extractive bioconversion provides a technique which integrates bioconversion and purification into a single step process. Extractive bioconversion of gamma-cyclodextrin (γ-CD) from soluble starch with cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) enzyme derived from Bacillus cereus was evaluated using polyethylene glycol (PEG)/potassium phosphate based on ATPS. The optimum condition was attained in the ATPS constituted of 30.0% (w/w) PEG 3000 g/mol and 7.0% (w/w) potassium phosphate. A γ-CD concentration of 1.60 mg/mL with a 19% concentration ratio was recovered after 1 h bioconversion process. The γ-CD was mainly partitioned to the top phase (YT=81.88%), with CGTase partitioning in the salt-rich bottom phase (KCGTase=0.51). Repetitive batch processes of extractive bioconversion were successfully recycled three times, indicating that this is an environmental friendly and a cost saving technique for γ-CD production and purification.

Matched MeSH terms: Phosphates/chemistry*
Characterization of bovine serum albumin partitioning behaviors in polymer-salt aqueous two-phase systems

Chow YH, Yap YJ, Tan CP, Anuar MS, Tejo BA, Show PL, et al.

J Biosci Bioeng, 2015 Jul;120(1):85-90.
PMID: 25553974 DOI: 10.1016/j.jbiosc.2014.11.021

In this paper, a linear relationship is proposed relating the natural logarithm of partition coefficient, ln K for protein partitioning in poly (ethylene glycol) (PEG)-phosphate aqueous two-phase system (ATPS) to the square of tie-line length (TLL(2)). This relationship provides good fits (r(2) > 0.98) to the partition of bovine serum albumin (BSA) in PEG (1450 g/mol, 2000 g/mol, 3350 g/mol, and 4000 g/mol)-phosphate ATPS with TLL of 25.0-50.0% (w/w) at pH 7.0. Results also showed that the plot of ln K against pH for BSA partitioning in the ATPS containing 33.0% (w/w) PEG1450 and 8.0% (w/w) phosphate with varied working pH between 6.0 and 9.0 exhibited a linear relationship which is in good agreement (r(2) = 0.94) with the proposed relationship, ln K = α' pH + β'. These results suggested that both the relationships proposed could be applied to correlate and elucidate the partition behavior of biomolecules in the polymer-salt ATPS. The influence of other system parameters on the partition behavior of BSA was also investigated. An optimum BSA yield of 90.80% in the top phase and K of 2.40 was achieved in an ATPS constituted with 33.0% (w/w) PEG 1450 and 8.0% (w/w) phosphate in the presence of 8.5% (w/w) sodium chloride (NaCl) at pH 9.0 for 0.3% (w/w) BSA load.

Matched MeSH terms: Phosphates/chemistry
Recovery of laccase from processed Hericium erinaceus (Bull.:Fr) Pers. fruiting bodies in aqueous two-phase system

Rajagopalu D, Show PL, Tan YS, Muniandy S, Sabaratnam V, Ling TC

J Biosci Bioeng, 2016 Sep;122(3):301-6.
PMID: 26922478 DOI: 10.1016/j.jbiosc.2016.01.016

The feasible use of aqueous two-phase systems (ATPSs) to establish a viable protocol for the recovery of laccase from processed Hericium erinaceus (Bull.:Fr.) Pers. fruiting bodies was evaluated. Cold-stored (4.00±1.00°C) H. erinaceus recorded the highest laccase activities of 2.02±0.04 U/mL among all the processed techniques. The evaluation was carried out in twenty-five ATPSs, which composed of polyethylene glycol (PEG) with various molecular weights and potassium phosphate salt solution to purify the protein from H. erinaceus. Optimum recovery condition was observed in the ATPS which contained 17% (w/w) PEG with a molecular weight of 8000 and 12.2% (w/w) potassium phosphate solution, at a volume ratio (VR) of 1.0. The use of ATPS resulted in one-single primary recovery stage process that produced an overall yield of 99% with a purification factor of 8.03±0.46. The molecular mass of laccases purified from the bottom phase was in the range of 55-66 kDa. The purity of the partitioned laccase was confirmed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

Matched MeSH terms: Phosphates/chemistry

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