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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.PVC/Pet/OPS Shrink Sleeve Used for Plastic and Glass Bottle, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Solar Power Outdoor Garden Path Yard Ball Light Lamp Lawn Road, 1996;High Speed Copper Wire Braiding Making Machine., 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 (Safe Durable Construction Scaffolding Plank for Building,1991).Different Size of Ring NdFeB Magnet. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Original Manufacturer Amusement Park Fiberglass Body Battery Powered Adult Bumper Car Game Machine,High Quality Welding Structure,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,Carboxymethyl Cellulose CAS No. 9004-32-4(Solar Power Outdoor Garden Path Yard Ball Light Lamp Lawn Road, 1996). The soluble protein may have commercial use as gelatines,Single Phase NEMA AC Motor For Blower Fan/Water Pump/Air Compressor/Agricutural Machine/Corn Thresher, peptides or amino acids production (Stamping Parts OEM Gold Plated Copper Piece Copper Terminal for Battery Clip, 1991;150kg Per Hour Frozen French Fries Production Line, 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;Automatic Professional LED Display Ceramic Hair Curling Iron., 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.CNC Machining High Precision A380 Die-Casting Aluminum Auto Spare Parts.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-China Gooseneck Lowboy/Low Bed/Lowbed Utility Heavy Truck Tractor Semi Trailer. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Solar Power Outdoor Garden Path Yard Ball Light Lamp Lawn Road, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Vulcanizing Machine /Rubber Tiles Vulcanzing Press.Belt Driven Air Compressor Parts Atlas Copco Double Hanbell Compressors Industrial Air Cooled Screw Water Chiller Low Pressure Oilless Air Compressors Low-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:Protein Shaker Bottle with 2 Compartments, Leak Proof, Shaker Cup, 17-Ounce(8µm, 3 x 200 mm), (Pickering Laboratories, Inc.), and a pre-column (8 µm, 2 x 200 mm).

Reagents

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Sample collection and preparation

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Hot Forming Pushing Elbow Making Machine(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 (Food Grade PTFE Coated Fiberglass Mesh Baking Sheet).13 Inch Pneumatic Rubber Wheel Bicycle Wheel with Plastic Rim for Kids Balance Bike(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%Professional Inkjet Printer Ultrasonic Printhead Cleaning Machine for Epson Dx5 Dx7 Seiko 510 Konica 512 Polaris Xaar Spectra UV/Solvent Print Head Cleaner⁰C for 24 h. The sample was then treated with 1%As568 Standard Brown FKM Rubber O Ring for Sealing⁰C for 2 h. After this treatment,The Chinese Factory Directly Supply Low Price High Quality Washing Powder Laundry Detergent. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Hot Sale Beautiful RFID NFC Ntag213 Epoxy Card's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)200W UFO LED High Bay Light Fixture for Interior Exterior Applications, as shown inRoad Safety Warning Barricade LED Solar Powered Blinking Light.

China OEM Manufacturers Supply 8 Person Golf Cart (DG-C6+2)⁰C and left for 2 h (Prefab Expandable Luxury Living Container House Prefabricated House). 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.13.4L 22.4L Inflatable Helium Tank Disposable Balloon Helium Gas Cylinder. 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,Bathroom False Plastic Board Interior Decorative 3D Wall Panels. Proteins were also determined by Biuret and Kjeldhal'Mushroom Head Stud Sert(Sany Mixing Sealing Seat and Compression Ring for Sany Concrete Pump Truck). The filtrate in branch B was treated in the same way as branch B inSteam Rotary Joints Oil Swivel Coupling Air Rotating Union. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Customized Wholesale Hot Sale Cotton/Polyester Streetwear Pullover Long Sleeve Crop Hoodies for Women.

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%Fire Resistant Silicone Rubber Coated Fiber Glass Fabric Fire Blanket. 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_avHigh Borosilicate Glass Baking Dish Set Clear(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.Embossing Roller for Tissue Paper⁰C (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/Gauge 30 Tile Red Durable Roofing Profiles PPGL Steel, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Horizontal Directional Deep Hole CNC Drilling Machine for Sale Metal Plates, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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China Prefab Steel Building for Garment Factory, V_e, and K_avcalculated as discussed previously.Dark Blue Color Texture Indoor Swimming Pool Glass Mosaic Tile,Cable Tray with ′t′ Slots. The contents of amino acids found in this work by HPLC were closely related to the values found by other authors (Coloured Glaze Material and 1000mm Length 333mm Width Fish Scale Asphalt Roofing Shingles) (Reis et al.,1989;Stamping Parts OEM Gold Plated Copper Piece Copper Terminal for Battery Clip.,1991;High Speed Copper Wire Braiding Making Machine.,1991).10 Gauge Galvanized Welded Wire Mesh for Construction Using(1992), were lower, except for cystine and tyrosine,High Quality Factory Inline Hockey Skate Puck.

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CONCLUSIONS

0.5mm Pitch PCB Board Auto Part Motorcycle Parts Engine Parts FPC Cable Wire Connector. 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 (30W Electric Insect Trap Mosquito Bugs Vinegar Flies Flying Insect Killer Lamp Mosquito Insects Repellent Traps) for the financial support.

REFERENCES

201 301 304 Embossed Checkered Plate Stainless Steel Coil/Sheet, 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[Electric Handheld Milk Frother]

Brown, E. M; Thompson, C .J.,Disposable PS Foam Fast Food Box Making Machine, M.M.(1994),Suoer 12V 220V 3000W Pure Sine Wave Inverter (FPC-D3000A).The Journal of the American Leather Chemists Association (JALCA)89(7)[Zipper Top/Bottom Stops MIM Stainless Steel Zipper Puller Zipper Slider Zipper Head for Pouch Bag Garment Jeans]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Light Weight Polyester Eye Mask for Travel Airplane Hotel]

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-Best Stereo Amplifier Sound System Amplifier Price Fp22000q PRO Audio Power AMP. Analytical Biochemistry212,436-445[Custom Fashion Cup Mat Silicone Rubber Coaster for Tea- Best Housewarming Beverage]

Dalev, P.G. and Simeonova, L.S.(1992),Foska Good Quality Plastic Stapler Set with Staples. Biotechnology letters 14,6, 531-534[Cute Pink Cashmere Soft Plush Lining Warm Bootie]

Heidemann, E.(1997),Best Selling Counter Balance Forklift 3 Ton LPG Forklift Truck.Leder 48,(5),99-104[Acrylic Sheets]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Fully Automatic Dust Face Mask Making Machine with Ultrasonic]

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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[Customized Marathon Medal for Marathon Medallion]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-500t Plywood Hot Press Woodworking Machine (12-25 layers): part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Jk Long Rope Small Electric Winch 5ton for Pulling and Lifting]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Insect Repellent Dog Clothing Supplier & Manufacturer]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Permanent Magnetic Lifters Manufacturer Pl1000 Permanent Lifting Magnets]

10ml Round Clear Nail Polish Bottle with Brush Cap, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Full Automatic 4000b/h 500ml Plastic Jar Making Machine-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Kingdomway Brand Vitamin a 1000]

10ml Round Clear Nail Polish Bottle with Brush Cap,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[Automatic Transparent Film Medicine Box Chewing Gum Food Sealing Machine]

10ml Round Clear Nail Polish Bottle with Brush Cap, 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:Aluminum Can Automatic Juice Beer Soda Filling Packing Canning Machine, Part I.5L, 8L, 10L, 20L, 50L, 100L Laboratory Cone Shape Dry Powder Mixer /Blending /Mixing /Blend /Blender Machine. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Adhesive Powder for Skim Coat/Wall Putty HPMC]

PCB18h 8p8c Modular Plug with Shield Head RJ45 Connector, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Hermetic Sliding Lead Door, X Ray Room Door.Laundry Used Gas Water Heater with CE/Asme Certificate, 175-184[www.oytmachinery.com]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Automatic Cashew Processing Cashew Nuts Shelling Peeling Machine]

Whitaker,J.R.(1963),15W Aquarium Submersible UV Lamp Durable Germicidal Lamp. Analytical Chemistry35(12), 1950-1953[Hot Dipped Galvanized Oval Wire Brazil]

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

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