One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite

Dimitrijević, Aleksandra; Veličković, Dušan; Bihelovic, Filip; Bezbradica, Dejan; Jankov, Ratko M.; Milosavić, Nenad

doi:10.1016/j.biortech.2011.11.077

Cited by 19 publications

(12 citation statements)

References 28 publications

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“…On one hand HAp is used by the body as a mineral reservoir owing to its ability to easily undergo ionic substitutions and accommodate a variety of differently sized and charged ionic species, while on the other hand HAp has been used with success as an adsorbent in chromatographic columns 50,51 , gas sensors 52 and environmental purification technologies 53,54,55 ; hence the rationale for testing both of these methods for the capture of selenite ions. Sorption of selenite proceeds by substituting phosphate groups on the surface of HAp 56 , yet in the case of co-precipitation the additional possibility for the bulk incorporation of selenite is likely to produce a higher ion incorporation efficiency.…”

Section: Resultsmentioning

confidence: 99%

One ion to rule them all: the combined antibacterial, osteoinductive and anticancer properties of selenite-incorporated hydroxyapatite

Uskoković

Iyer

2017

J. Mater. Chem. B

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Although hydroxyapatite (HAp) has been doped with dozens of different ions, the quest for an ion imparting a combination of properties conducive to bone healing is still ongoing. Because of its protean potency and the similarity in size and shape to the phosphate tetrahedron, selenite ion presents a natural ionic substitute in HAp. The incorporation of selenite into synthetic HAp using two different methods – co-precipitation and ion-exchange sorption - was studied for its effect on crystal properties and on a triad of biological responses: antibacterial, anticancer and osteoinductive. Co-precipitation yielded HAp with higher selenite contents than sorption and the stoichiometry of HAp richest in selenite was represented as Ca9.75(PO4)5.75(SeO3)0.25(OH)1.75. Crystallinity of HAp decreased in direct proportion with the amount of selenite incorporated. Because of their lower selenite content, HAp powders prepared by ion-exchange exhibited a consistently higher crystallinity compared to the co-precipitated ones. Annealing partially recovered the crystallinity, yet the difference in crystallinity between powders prepared by co-precipitation and by ion-exchange remained, suggesting that the amorphization is mainly due to structural incorporation of selenite, not its effect on the crystal growth kinetics. The addition of selenite changed the morphology of HAp nanoparticles from acicular to rounded and affected the crystal lattice parameters in different ways depending on whether the powders were annealed or not. As for the annealed powders, the incorporation of selenite contracted the lattice in both a and c crystallographic directions. In the agar diffusion assay, the effectiveness of HAp was more dependent on the presence or absence of selenite in it than on its concentration and was highest against E. coli and S. aureus, moderately high against S. enteritidis and ineffective against P. aeruginosa. In liquid inoculation tests, on the other hand, the antibacterial activity of HAp was directly proportional to the amount of selenite contained in it. The viability of K7M2 osteosarcoma cells decreased in direct proportion with the amount of selenite in HAp and was significantly different from the untreated control and from pure HAp at contents equal to or higher than 1.9 wt.%. In contrast, no reduction was observed in the viability of primary fibroblasts treated with HAp incorporating different amounts of selenite ions, suggesting their potentially selective anticancer activity: lethal for the cancer cells and harmless for the healthy cells. Finally, mRNA expression of bone gamma-carboxyglutamate protein (BGLAP3) was higher in differentiated MC3T3-E1 osteoblastic cells treated with selenite-incorporated HAp particles than in cells treated with pure HAp. The osteoinductive effect was due to an overall higher metabolic activity of cells treated with the particles and not due to increased proliferation. In such a way, a triad of antibacterial, osteoinductive and anticancer activities was attributed to selenite-incorporated ...

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Section: Resultsmentioning

confidence: 99%

One ion to rule them all: the combined antibacterial, osteoinductive and anticancer properties of selenite-incorporated hydroxyapatite

Uskoković

Iyer

2017

J. Mater. Chem. B

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“…Most purification processes employ concentration step, followed by chromatographic separation which can be single or multiple. Most of the conventional purification attained through multistep processes has been found to yield 2-20% of enzyme [4]. For large scale application purification strategies are required which are fast, inexpensive, high yielding and simple in operation.…”

Section: Introductionmentioning

confidence: 99%

Biochemical Characterisation of a Newly Isolated Low Molecular Weight Lipase from Rhizopus oryzae NRRL 3562

Adak¹,

Banerjee²

2013

Enz Eng

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“…Lipases from the Candida genus, including those from Candida rugosa, Candida antarctica, Candida cylindracea, present molecular weight ranging from 33 to 64 kDa and multiple isoforms [13][14][15][16]. These lipases normally show similar biochemical properties such as optimal activity in the pH range 7.0-8.0 and at 30-60°C [13][14][15][16]. Lipases from C. antarctica (CALB), C. rugosa and Mucor miehei were immobilized by adsorption on hydrophobic supports as a quick and inexpensive alternative to obtain purified samples, rendering high yield and, in some cases, activation of the enzyme [17].…”

Section: Introductionmentioning

confidence: 99%

Biochemical properties of free and immobilized Candida viswanathii lipase on octyl-agarose support: Hydrolysis of triacylglycerol and soy lecithin

Almeida

Terrasan

Terrone

et al. 2018

Process Biochemistry

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A B S T R A C TMicrobial lipases are important enzymes in food and pharmaceutical industries. In this work, a Candida viswanathii lipase was purified by hydrophobic interaction chromatography on octyl Sepharose. The purification presented 78.4% yield and the enzyme was 8.7-fold purified, with specific activity 700.4 U/mg protein and 69 kDa molecular weight. Immobilization of the enzyme on the same support presented 72.5% yield and a derivative with 101% expressed activity (109.2 U/g support), indicating hyperactivation of the enzyme. Optimal activity for both free and immobilized lipase was observed at pH 4.0 and 45°C. The free and immobilized lipase showed broad-range stability from acid to neutral pH, and apparent activation and stability on organic solvents. The derivative was 60-fold thermostabilized in relation to the free enzyme and fully retained its activity after four cycles of p-nitrophenyl palmitate hydrolysis. Slight activation was observed with dithiothreitol and βmercaptoethanol. The free and immobilized lipase efficiently hydrolyzed monoesters, simple and mixed long chain triacylglycerols, as well as soy lecithin. The activity and stability in acid pH, the organic solvent tolerance and the lecithin hydrolysis indicate high potential application of the enzyme and its derivative in textile, food and pharma industries and for chemical synthesis.

show abstract

One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite

Cited by 19 publications

References 28 publications

One ion to rule them all: the combined antibacterial, osteoinductive and anticancer properties of selenite-incorporated hydroxyapatite

One ion to rule them all: the combined antibacterial, osteoinductive and anticancer properties of selenite-incorporated hydroxyapatite

Biochemical Characterisation of a Newly Isolated Low Molecular Weight Lipase from Rhizopus oryzae NRRL 3562

Biochemical properties of free and immobilized Candida viswanathii lipase on octyl-agarose support: Hydrolysis of triacylglycerol and soy lecithin

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