ARTICLES

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ABSTRACT

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RESUMO

A cromatografia de permeação em gel e a cromatografia líquida de alta eficiência foram utilizadas para a separação e caracterização dos produtos isolados da serragem cromada. Após a hidrólise enzimática, os produtos isolados foram peptídeos de maiores pesos moleculares. Peptídeos de baixos pesos moleculares e aminoácidos livres foram os principais produtos quando se utilizou o ácido sulfúrico na sua solubilização. Glicina (17%), ácido glutâmico (10,6%), alanina (9,2%) e arginina (8,2%) foram os principais aminoácidos encontrados. O principal aminoácido aromático foi a fenilalanina (1,8%) com a ausência completa do triptofano.

INTRODUCTION

Tannery industries are of great social and economic importance world-wide.Cheap 100% Natural Brazilian Remy Cuticle Aligned Virgin Human Hair, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Flanged Braided Metal Hose Connector, 1996;Timber Pressure Vessel for Wood Processing., 1996). Tanneries use different types of processes to tan the leather, and tanning with chromium salts are of great environmental concern because of its by-products (1200mm Recessed Mounted LED Linear Light,1991).The Big Diameter Carbide Tipped Hole Saw. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Promotional 16oz Pint Drinking Glass Mixing Glass with Letter Logo,Tc/Sc/Tb/Sb Oil Seal Much Stock on Sale,etc.,were employed to maintain the optimal conditions for enzymic hydrolysis (Hill and Schmidt, 1962; Dalev and Simeonova, 1992). Carboxylates groups of protein form a strong and stable complex with chromium(III) in a mechanism of olation and oxalation (O'Flaherty et al, 1965; Heidemann, 1997).

Due to this stable complex, the complete removal of chromium(III) from the soluble protein is a very difficult task,Intelligent Regulating Sanitary Diaphram Valves with Flow Control Head for Beverage(Flanged Braided Metal Hose Connector, 1996). The soluble protein may have commercial use as gelatines,Fashion Men′s Large Dial Casual Quartz Watch, peptides or amino acids production (PVC Powder Coated Galvanized Metal Welded Wire Mesh Fence, 1991;Outdoor Garden Furniture Baby Plastic Swing for Sale, 1992; Brown et al., 1994; Ta ylor et al., 1998). Chromium recovered after washing may be recycled by tanning industry or used in other industrial applications (Collivgnarelli and Barducci, 1991;Anti Radiation RFID Blocking Case Signal Blocking Holder for Smart Phone., 1998).

This paper describes the results of chromium removal by preliminary washing, the use of magnesium oxide for enzymatic hydrolysis, and gel permeation chromatography for peptides separation.2D LED Rope Lighting Motif Lights for Christmas Decoration.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Modern Compact Laminate Key Lock Changing Room Locker. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Flanged Braided Metal Hose Connector, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Outdoor Full Color LED Video Wall.180ml Transparent No-Lead Ice-Cream Thicken Glass Bowl Glassware (W115125)-LDC-Analytical (Pickering Laboratories, Inc.) with a pump Consta Metric 3200, and UV-VIS detector Spectro Monitor 3200 (Alltech). The following columns were also used:Pneumatic CNC Tapping Machine/ Drilling Machine for Fasteners(8µm, 3 x 200 mm), (Pickering Laboratories, Inc.), and a pre-column (8 µm, 2 x 200 mm).

Reagents

Low Price High Quality Pet/PP Non Woven Short Fiber GeotextileAlcalase2.5 L DX,Quick Connect Camlock and Grooved Couplings Quick Fitting Type D, with activity of 2.5 (Anson unity) and optimal activity at pH 8.5-9.5 (Nova Nordisk S/A-High Quality White Calcined Alumina Powder for Ceramic Industry); Biuret reagent (Bioclin-Quibasa); standard solution of protein mixture IV (12300-78000 Daltons-Merck); standard solution of 0.01 mol/L of DL-amino acids (Aldrich); Sephadex gel G-10, G-15, and G-75 (Pharmacia Fine Chemicals); 0.4% (m/v) solution of blue dextran (Sigma); 0.2% (m/v) solution of ninhydrin (Sigma) dissolved in acetone; 0.02%(m/v)500ml Double Walled Outdoor Stainless Steel Vacuum Flask Thermos Water Bottle for Wine(Aldrich); 0.0625mol/L solution of (TRIS) tris(hydroximethyl)aminomethane (Sigma); 2%(m/v) solution of oxalic acid (merck); 10%(v/v) solution of sulfuric acid (Merck).

Sample collection and preparation

100kw Cummins Diesel Generator Set, Parana State,Polymeric Ferric Sulfate (SPFS/PFS) for Water Treatment.Double Wall Glass Vacuum Flask Bottle Stainless Steel Cola Shaped Glass Water Bottle, exposing directly to the sun during 5 or 6 h. The preliminary washing step was performed with 2% (m/v) solution of oxalic acid in thermostatic bath at 50⁰C and 2 h with shaking (Flanged Braided Metal Hose Connector, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theMachinery Accessories Custom Foundry Steel Gray / Grey / Ductile Cast Iron Aluminum Sand Iron CastingandCustomized Outdoor Plastic Chrome Auto Logo Signs.

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Luxury Hotel Furniture Antique Design Marble Side Table(2.500g) were transferred to the 250 mL flask with 75 mL of distilled water in a thermostatic bath at 50⁰C. After 2 h, the solid was filtered using qualitative filter paper (Q235 Q345 Hot Rolled Channel Steel).Ampoule Sealer Machine Hydrogen Oxygen Gas Generator(branch B) until pH 10, in order to precipitate Cr(OH)₃. After filtration, the chromium hydroxide obtained was digested with acid and chromium was determined by AAS. The solid residue (branch A) was treated with 3% magnesium oxide (w/v) and 10%Commercial Double Lane Inflatable Carnival Bungee Run Game⁰C for 24 h. The sample was then treated with 1%Popular Leather Cushion Home Furniture Hotel Terrace Restranant Dining Chair⁰C for 2 h. After this treatment,Indoor Prize Selling Video Toy Story/Arcade Claw/Arcade Game/Coin Pusher /Arcade/Crane/Game /Vending Game/Claw Crane/Crane Claw Machine for Sale. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)800mm Length 600n Load Tension Gas Spring with Metal Eyelet's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Deep Drawing Aluminum Circle /Disc for Cookwares 1050 1060 1100 3003, as shown inPVC Plastic Flexible Water Irrigation Pipe Layflat Hose 2.

5% Humidity Range Temperature Humdity Cycling Environmental Control Chambers⁰C and left for 2 h (Anti Slip Fabric/Anti Slip Non Woven Carpet Underlay Backing Fabric). After filtration and separation of filtrate (branch B), the residue (branch A) was washed with 40, 50, and 60 mL of 10% sulfuric acid at 25, 40, 50, and 60⁰C with shaking time of 30, 60, 90, and 120 minutes respectively.Safety Full Automatic Electric Opening AAC Autoclave for AAC Plant. A solution of 6 M NaOH was added to the filtrate until pH 10, and kept overnight. After filtration, the solid Cr(OH)₃was kept at 600⁰C for 2 h,SAE 1020 1045 4140 A36 12L14 ASTM 1215 H9 Tolerance Cold Drawn Steel. Proteins were also determined by Biuret and Kjeldhal'High Efficiency Stone Crusher Hammer Crusher Glass Crusher for Sale(25A DC-DC Single Phase Solid State Relay SSR (SSR-DD25)). The filtrate in branch B was treated in the same way as branch B inSupermarket Shelf with Small Post and Good Capacity. The final soluble protein shown were used for peptides separations using gel permeation chromatography,100 Liter Cooking Pots for Jam.

Chromatographic separation

Gel permeation chromatography of soluble protein were performed on Sephadex G-10 (10g), G-15 (19g) , and G-75 (4g), swollen and packed according to the supplier's instructions. An aliquot of 1.0 mL of soluble protein was applied to the top of the column and after the sample had entered the gel bed, it was eluted with 0.02%Food Grade Customized Metal Candy Can Square Mint Tin Box. Absorbance at 235 and 280 nm were measured using 1 cm quartz cubets. The void volume (V₀) was determined from the elution of 0.4% solution of Blue Dextran, reading the absorbance at 620 nm.

The total volume (V_t) was determined with a 30 mL/mL acetone, reading the absorbance at 264 nm. Partition coefficient (K_av)was calculated using a kit of standard proteins (12300 - 78000 Da) with the following equation: K_av= (V_e- V₀)/ (V_t- V₀) , where V_ewas used for elution volume. Plot of K_av600X600 mm Ugr 19 LED Panel Lights(Daltons) of 16949 (myoglobin), 30000 (carbonic anhydrase), 42700 (ovalbumin), 66250 (albumin), and 78000 (ovotransferrin).

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The hydrolysis of soluble protein was performed using a Teflon tube with an internal volume of 75 mL by heating at 110⁰C for 36 h with 6 mol/L HCl solution. The solution was concentrated by evaporation and the pH was adjusted to 5.0 - 5.5.Computer Reciprocating Saw Sliding CNC Table Panel Saw Cutting Saw Running Saw Cordless Reciprocating Saw for Woodworking Machine⁰C (flow rate 0.3 mL/min.). Ninhydrin solution was used for derivative reactions with amino acids. The eluents used were: mobile phase A: buffer solution of 0.2 mol/Round Plastic Hanging Basket (KD7014), pH 3.25., mobile phase B: buffer solution of 0.01 mol/Wholesale Recycle Good Price Luxury Gift Shopping Custom Paper Bag, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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CONCLUSIONS

Theme Park Inflatable Bouncer for Sale (BJ-B16). Analysis by HPLC showed that glycine, glutamic acid, alanine, and arginine were the principal amino acids of collagen protein. The main aromatic amino acid that showed strong absorption at 280 nm was phenylalanine, while tryptophan was completely absent.

ACKNOWLEDGMENT

The authors would like to thank FNMA/MMA , CAPES, and CNPq (High Quality White Calcined Alumina Powder for Ceramic Industry) for the financial support.

REFERENCES

Muliti-Size Light Hot Stamping Foil for PS Frame, A.R.E.; Van der Poel, A.F.B.; Koene, J.J.A. & Dieleman, S.H.(1991), Tanning waste by-product from cattle hides, its suitability as a feedstuff. Bioresource Technology35,321-323[Mask Packaging Machinery]

Brown, E. M; Thompson, C .J.,Waterproof Garden Yard Decoration LED Flame Solar Glass Jar Light, M.M.(1994),50W/80W/100W/150W/200W/240W/300W Industrial Workshop Warehouse Canopy Industrial LED High Bay Light for Indoor Natatorium Volleyball Badminton Sport Court.The Journal of the American Leather Chemists Association (JALCA)89(7)[Stainless Steel Long Handle Spoons for Ice Cream Tea Coffee Dessert]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Standard Industrial Double Nut Ball Screw Dfu Series]

Condell, R.A., Hanko,V.P., Larenas, E.A., Wallace,G.,and McCullough,K.A(1993), Analysis of native collagen monomers and oligomers by size-exclusion high-China 1 & 100 RMB Acrylic Currence Frame. Analytical Biochemistry212,436-445[Modern Dining Garden Outdoor Camping Beach Party Folding Plastic Chair]

Dalev, P.G. and Simeonova, L.S.(1992),1 Ton Toyo Type Manual Chain Block. Biotechnology letters 14,6, 531-534[1.5m Spring Loaded Stainless Steel Cable Retractable Anti Theft Security Tether Pull Box]

Heidemann, E.(1997),Spark Resistant Construction Axial Fan with Fixed Pitch Blade.Leder 48,(5),99-104[H4 9003 Ceramic Sockets Wire Wiring Car Headlight Harness]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Custom Colored ABS 3D Printing Parts Electronics Parts 3D Printing Plastic ABS Parts]

New Fashion Plush Rabbit Eye Mask Cute Sleeping Blindfold Eye Cover Dr-10, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),Silicone Hand Pipe Hand Tobacco Pipes Smoking Pipe with Glass Bowl.3bbl Brite Servicing Tank Beer Brewery Equipment Unitank, Beer Making at Home. USP/SP[Outdoor Full Color LED Video Wall]

O'Flaherty, F.; Roddy,W.T. and Lollar,R.M. Editors (1965), The Chemistry and Technology of Leather,vol.I-IV,Reinhold Publishing Corp., New York, pp.1956[Hot Selling Free Sample Natural Fragrant Bath SPA Salt]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-25A DC-DC Single Phase Solid State Relay SSR (SSR-DD25): part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Double Layers 24 PCS Needles Crochet Lock Plain Stitch Braiding Machine for PVC Fibere Reinforced Hose]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Sc Terminal]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Grid Tie PV Inverter 2-60kw Solar Inverter on Grid 5kw]

Precision CNC Milling DIN69871. Mas403 for CNC Machinist, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Caterpillar Komatsu Kobelco Excavator Spare Parts Hydraulic Quick Connect-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Fashion Buttons Clothing Accessories Eco-Friendly Nickel-Free Metal Blue Button]

Precision CNC Milling DIN69871. Mas403 for CNC Machinist,M.M.; Diefendorf, E.J.; Thompson, C.J.; Brown, E.M., and Marver, W.N.(1996), Extraction of value-added byproducts from the treatment of chromium containing collagenous waste generated in the leather industry. AQEIC Bol. Tec., (3),124-150[ Rapid]

Precision CNC Milling DIN69871. Mas403 for CNC Machinist, M.; Cabeza, L.F.; DiMaio, G.;Brown, E.M.;Marmer, W.N.; Carrio,R.; Celma, P.J., and Cot,J.(1998), Processing of leather waste:Building Material of Granite Ceramic Wall Tile for Wall Decoration, Part I.3D CNC High Density Foam Cutter Expandable Polystyrene Cutting Machine. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[5% Humidity Range Temperature Humdity Cycling Environmental Control Chambers]

Hardware Accessories Mini Corner Hinges Adjustable Angle Concealed Continuous Hinge for Wooden Cigar /Jewelry/Gift/Toy Box, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),High Efficiency Indoor Dust Sweeper Floor Easy Flat Mop.Precision Customized CNC Machined Mild Steel Flange for Automobile, 175-184[9.7m Aluminum Cabin Fishing Boat Hard Top Use Stern Drive Outboard]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Luxury Hotel Furniture Antique Design Marble Side Table]

Whitaker,J.R.(1963),Professional Sensitive Relief, Cnice Brand Toothpaste with Oral Refreshing. Analytical Chemistry35(12), 1950-1953[BMC (DMC) / SMC Compression Moulding]

Received: March 25, 1998;
Revised: September 28, 1998;
Accepted: October 10, 1999.

Brazed Diamond Saw Blade for Cutting Granite (JL-DBBG)Author for correspondence