S458 Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576

[P-M.86]

Production of alginate spherical particles for delivery of barkcatechins

Hugo Pulgar, Veronica Sanhueza, Estrella Aspé, Marlene Roeckel,Katherina Fernández ∗

Universidad de Concepción, ChileKeywords: Bark; Catechins; Delivery; Pinus radiata

Introduction: In Chile, Pinus radiata is the main tree used in woodindustry; it generates amounts of bark residues, a byproduct with ahigh amount of natural polyphenols like proanthocyanidins (PAs).PAs are very unstable in human systems; they must be protectedto preserve their bioactivity if they want to be used as food sup-plement. The objective of this work was to encapsulate pine barkextract into alginate spheres, and to evaluate the phenols, tanninsand anti-radical capacity of this extract as well as the entrapmentefficiency of the encapsulation process.

Methodology: Phenols from pine bark were obtained usingmicrowaves with acetone/MQ water (7:3%v/v) and the extract waslyophilized to generate an impure extract (IE), and four differentfractions (F1 to F4) purificated by size exclusion chromatography.Total phenols (by Folin-Ciocalteau), tannins (by methyl-celluloseprecipitation) and anti-radical capacity (by DPPH radical method)were analyzed; these results were compared with BHT andPycnogenol®, commercial antioxidants under the same concen-trations. IE was trapped using a calcium alginate gel with twoconcentrations of CaCl2 (0.05 and 0.01 M) and different immersiontimes to find the maximum level of phenols, which was determinedafter dissolve the capsules with a sodium citrate solution.

Results and discussion: The results showed that F1 and F3-F4 hadthe highest level of phenols and tannins, respectively (Table 1); and

Table 1Concentration of phenols, tannins and anti-radical capacity as inhibition percentage(12 �g extract/mL) for IE and fractions purified.

Sample Total phenols (geq.C/g extract)

Tannins (mgeq.C/g extract)

Inhibitionpercentage (%)

IE 5,2 ± 0,12ad 576 ± 4a 82,3 a ± 2,2aF1 6,9 ± 0,25b 476 ± 14b 68,4 b ± 4,1bF2 5,5 ± 0,03a 523 ± 5c 48,8 c ± 0,7cF3 6,4 ± 0,25c 859 ± 7d 64,2 b ± 3,4bF4 5,1 ± 0,06d 850 ± 8d 79,5 a ± 1,2aBHT - - 52,5 ± 1,3Pycnogenol® - - 16,3 ± 1,5

the highest anti-radical capacity was founded for IE and F4. All theextracts had higher anti-radical capacity than the two commercialsantioxidants tested, except F2. There were not a correspondenceamong a higher level of phenols and the anti-radical capacity.

The encapsulation efficiency was higher at lower concentrationof CaCl2 and minimum immersion time in the solution, reaching amaximum of 47.35% of entrapment efficiency.

Conclusions: Results show that pine bark has high anti-radicalcapacity and alginate has an elevated capacity of trapping tanninsfrom the extract.

doi:10.1016/j.jbiotec.2010.09.674

[P-M.87]

High throughput metabolomics analysis in normal and K-Rastransformed murine fibroblasts

Daniela Gaglio 1,2,∗, Christian Metallo 2, Ferdinando Chiaradonna 1,Chiara Balestrieri 1, Lara Sala Danna 1, Gregory Stephanopoulos 2,Lilia Alberghina 1, Cristina Messa 2

1 Department of Biotechnology and Biosciences, University of Milano-Bicocca, Italy2 Department of Chemical Engineering, Massachusetts Institute ofTechnology, United StatesKeywords: Cancer; K-Ras; Nutrient; Omics Technologies

Transcriptomic analysis is an essential tool to study gene expres-sion and identify the genes involved in human diseases as cancer.However, this important approach is insufficient to achieve adetailed description of changes in biochemical regulation elicitedby a given perturbation or by a pathological condition as the caseof cancer cells. Numerous studies have indicated that tumorigene-sis is associated with several metabolic alterations, resulting fromboth transcriptional and post-translational events, that activelyparticipate to the onset and maintenance of transformation pro-cess. Therefore, to identify and characterize specific metabolicalterations, able to promote growth of K-ras transformed tumorcells as compared to normal counterpart, we show the integrativeresults of data generated by high-throughput techniques, transcrip-tomic and metabolomic, upon alterations in nutrient availability.In particular, transcriptomic data show in transformed cells anincreased expression of several glycolytic genes as well as a dereg-ulated expression of both positive and negative regulators of TCAcycle. In addition, they show an increased expression of glutaminemetabolism genes whose encoded product sustain amino acids,nucleotides and glutathione synthesis using glutamine as a keyintermediate. These results have been confirmed by GC/MS anal-ysis following stable-isotope -[13C]Glucose, [13C]Glutamine and[15N]Glutamine- labeling. Indeed, K-ras Transformed cells show,in keeping with transcriptomic data, an increased glucose to lac-tate conversion, a decreased TCA cycle flux and a preferentialutilization of glutamine-derived carbon and nitrogen for glu-tathione, amino acids and nucleotide synthesis. Together thesedata indicate that transformed cells use glucose and glutamineas a main source of energy and anabolic precursors respectivelyand offer a more global view of cancer cell metabolism that canbe potentially exploited to the development of new cancer thera-pies.

doi:10.1016/j.jbiotec.2010.09.675

[P-M.88]

Bolinaquinone, a New Clathrin-Mediated Endocytosis Inhibitorby Chemical Proteomics

L. Margarucci ∗, M.C. Monti, B. Fontanella, R. Riccio, A. Casapullo

Università di Salerno, ItalyKeywords: Marine Natural Product; Chemical Proteomics; MassSpectrometry

Although the therapeutic potential of the most promising com-pounds is being evaluated in preclinical and clinical trials, oftentheir intracellular targets and the interaction profile remain largelyunknown. In recent years, mass spectrometry-based chemical pro-teomics approaches have been applied to the macromoleculartarget discovery and to the characterization of drug–target inter-