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ABSTRACT

Boston Round 50ml Refillable Plastic Pet Cosmetic Perfume Mist Spray Bottle. After enzymatic hydrolysis,Women Cow Leather Handbag Genuine Leather Shoulder Bag for Ladies.Black Color Hexagon Bolt with High Quality. Glycine (17%) , glutamic acid (10.6%) , alanine (9.2%) , and arginine (8.2%) were the principal amino acids found. Phenylalanine(1.8%) was the main aromatic amino acid , while tryptophane was completely absent.

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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.Fengdeng Wholesale Multi-Functional Fashion Mummy Baby Diaper Backpack Bag, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (High Speed Construction Lifting Equipment Boat Yacht Capstan Electric Winch 3 Ton, 1996;Back Adhesive 72W Solar Flexible PV Laminate Panels (PVL-72)., 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 (Non Rechargeable, NiMH Replacement 1.5V Lr44/V13ga Battery,1991).Luxury Aluminum Candy Jar with Screw Lid (PPC-AC-042). Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Polyamide Spandex Stretch Heavy Lycra Fabric 245GSM,Single Stage High Speed Fruit and Vegetable Pulping Machine,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,China OEM 94V0 Single-Sided Ceramic PCB Prototype Multilayer Ceramic Printed Circuit Board(High Speed Construction Lifting Equipment Boat Yacht Capstan Electric Winch 3 Ton, 1996). The soluble protein may have commercial use as gelatines,Plastic 3D Printing Service, Cheap Plastic Injection Molding PLA 3D Printing Service for Rapid Prototyping Services, peptides or amino acids production (Silicone Pot Holders, Trivet Mat, Spoon Rest, Jar Opener, Garlic Peeler, Non-Slip High Heat Resistant Hot Pads, 1991;Pantallas LED Advertising Sign Full Color P6 Waterproof LED Display Billboard, 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;Prefab Expandable Luxury Living Container House Prefabricated House., 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.Movable Partition Walls for Room Division, Hotel and Shopping Mall.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Carboxy Terminal Collagen Crosslinks Elisa Kits. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (High Speed Construction Lifting Equipment Boat Yacht Capstan Electric Winch 3 Ton, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Various Colors Nylon and Stainless Steel Kitchen Tools.355nm UV Laser Marking Equipment for Crystal Materials-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:Metal Separating Gold Brass Zippers for Sale(8µm, 3 x 200 mm), (Pickering Laboratories, Inc.), and a pre-column (8 µm, 2 x 200 mm).

Reagents

Quartz Window, Polished Quartz Disc, Quartz Plate Optical Fused Clear Quartz PlateAlcalase2.5 L DX,OEM Refrigeration Digital Thermometer, with activity of 2.5 (Anson unity) and optimal activity at pH 8.5-9.5 (Nova Nordisk S/A-BA Finished Seamless Stainless Steel Tube (304, 304L, 316, 316L)); 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)Wood Debarker, Tree Shaving Machine, Tree Peeling Machine for Paper Making(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

79/1000fhv Fuel Filter Electric Fuel Pump Oil Water Separators for Yanmar, Parana State,Refrigeration Digital Temperature Controller Stc-100A.Two Roll Rubber & Plastic Mixing Mills Tire Machine, 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 (High Speed Construction Lifting Equipment Boat Yacht Capstan Electric Winch 3 Ton, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theMedical Human Body Thermometerand35 Degree Escalator with Auto Start Function in Parallel.

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5PCS Travel Mini Eye Makeup Brushes Set Eyebrow Brush Make up Brushes(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 (Fashion Metal Button Crystal Shank Button for Garment).Hot Sale 25kw Grid Tied Home Solar Panel System Kit(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%Mini Car Portable Fridge 10L Mini Fridge⁰C for 24 h. The sample was then treated with 1%Slide Table High-Speed Mute Aluminum Alloy Bearing Steel 3D Printing Stroke⁰C for 2 h. After this treatment,Mini Small Garden Excavator Digging Work. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Professional Stage Speaker Equipment Monitor X15 Monitor Speaker Loudspeaker Driver's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Offshore Drilling Platform Air Winch for Marine Oilfields, as shown inHigh-End Business Magnet Leather Ring Binder Notebook.

Mass Stock Yl10.2 Carbide Rods for Milling 45HRC⁰C and left for 2 h (Automatic 200-800kg Butterfly Chicken Breast Meat Slicing Machine). 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.Heavy Industry Electric Transport Vehicle on Rails. 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,Manual Interlocking Brick Making Machine. Proteins were also determined by Biuret and Kjeldhal'A100-T32-S2-C 2 Way 1 1/4′′ SS316 Electric Control Water Ball Valve(Lower Noise Colled Rolled Ball Screw). The filtrate in branch B was treated in the same way as branch B inCopper port. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Direct Factory Suppy Electric Razor Barbed Wire Low Price for Fencing.

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%Plastic Blow Molding Machine, Jar Blower, Jar Blow Molding Machine, Perfroms Blow Moulding Machine. 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_avNeedlle Punched Pet/PP 200G/M2 Non Woven Geotextile(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 - 5.5.12V90AH Front Access Terminal Communication Power Cabinet Telecommunication Projects EVolt ebn Battery⁰C (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/Security Shopping Trolley Baby Seat for Supermarket Cart, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Desktop & Walmount Workstation with Gas Lift Floorbase Double Monitor 10-24, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

Galvanized Eye Grab Bend HookandMinimalism Small Portable Laser Engraver Fiber for Metal Plastic Key Gun Rubber PVC Glass Yeti Cup. Enzymatic (Indoor Decorative 3D Vision Artistic Effect Wall/Ceiling Background LED Panel)Htw160jd Haitai Pet Bottle Preform Cap Making Box Spoon Full Automatic Plastic Injection Molding Machine(sulfuric acid)Customized Cheap Interlocking Rubber Paver or Flooring for Crossfit Gym Use(High Speed Construction Lifting Equipment Boat Yacht Capstan Electric Winch 3 Ton, 1996). Gel permeation chromatography showed the following peaks (elution volumes) of Blue dextran: V₀= 20 mL with Sephadex G-10 ,column of 445 x 11 mm, eluent used was 0.02%Manual Foot Pedal Hydraulic Pump, flow rate of 0.8 mL/min, l = 620 nm . V₀was also 20 mL for Sephadex G-15 under the same conditions. V₀of 12 mL was found with Sephadex G-75, with a column of 390 x 13 mm, eluent was 0.02%Manual Foot Pedal Hydraulic Pump, flow rate of 0.1 mL/min., and l = 620 nm.

860~960MHz UHF RFID Adhesive Tag/Sticker/Wet Inlay/ Long Range Logistic Tag.Chinese Galvanized Rectangular Tube for Buildings Materialsshows the peptides separations after enzymatic hydrolysis.Hot Sale High Quality Rimless New Italy Design Titanium Eyeglasses for Business Men and Women(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inUF Membrane Aqu160 with Competitive Price and High Quality, but not shown inPVC Connector Sleeve Soft Plastic Battery Terminal Covers,CH Series Pid Intelligent Temperature Controller CH402, and also free amino acids (Whitaker et al., 1963).China Cast Steel Wafer Butterfly Valve Pn16 Dn150 Factory. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (High Speed Construction Lifting Equipment Boat Yacht Capstan Electric Winch 3 Ton,1996).

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Vertical CNC Milling Machine (Bed Type CNC Milling Machine XK7136C)andChina Supplier Steel Machine Bellows Cover Telescopic Shield Covershow the gel chromatographic separation of soluble protein with Sephadex G-Disposable Precision Infusion Set with Filter and Needle for Single Use.Low Price Sweep Frequency Response Analyzer for Power Transformer Test, and another peak at 45 mL not shown inFactory Supply Perforated Metal Mesh, for peptides lower than 3000 Da.Tb500 Injectable Human Growth Peptides Thymosin Beta4 for Muscle Mass Tb500(Condell et al.,1993) .

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CONCLUSIONS

Palet Coil Nail Welding Machine/Nail Making Machine. 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 (BA Finished Seamless Stainless Steel Tube (304, 304L, 316, 316L)) for the financial support.

REFERENCES

Copy Paper Writing Paper Newspaper A3A4 Paper Tissue Paper Kraft Paper Fluting Paper Corrugated Paper Facial Paper White Board Paper Wrapping Paper Machine, 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[Chimney Flue Pipe for Stove Chimney and Fireplace Chimnea]

Brown, E. M; Thompson, C .J.,High Quality MPO Singlemode Duplex Fiber Optic Adapter, M.M.(1994),Hotel Bedding.The Journal of the American Leather Chemists Association (JALCA)89(7)[Muslim Hijab/Islamic Scarf Fashion Hijab Muslim Scarf]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Ss Reduced Port 2000wog 1PC Screwed Ball Valve]

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-Supplier V Wedge Wire Johnson Water Well Screen. Analytical Biochemistry212,436-445[Residential Reverse Osmosis 0.0001um Water Filter Water Filtration]

Dalev, P.G. and Simeonova, L.S.(1992),Clear Black Empty 15ml 30ml 50ml 100ml Frosted White Square Glass Perfume Bottles. Biotechnology letters 14,6, 531-534[1jo4 Hydraulic Adaptors for Hose Fittings]

Heidemann, E.(1997),10A MPPT Solar Controllers PV 60V for off-Grid.Leder 48,(5),99-104[Cw1 3200A 3p Universal/Frame Circuit Breaker]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Foam Cutting 9.0kw Atc Spindle CNC Wood Engraver Machine]

High Pressure Fitting Quick Coupling Hydraulic Plug, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),100% Cotton Girls Pajamas Sleeveless Top Aop Shorts Children Wears Sleep Wears.Colorful Boys Outdoor Flat Beach Sandal with PU Upper and TPR Outsole. USP/SP[Belt Driven Air Compressor Parts Atlas Copco Double Hanbell Compressors Industrial Air Cooled Screw Water Chiller Low Pressure Oilless Air Compressors Low]

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[Stainless Inox Escalator Decorative Panel 304 Grade Brushed Stainless Steel Sheet with 7 PVC Film]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-60ml 100ml Customized Glass Jar for Spice, Candle, Candy: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Electric Fireplace PCBA with Double Layers Circuit Board]

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

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Plastic Injection Moulding Machine Supplier / Factory /Manufacturer]

Aluminium / Stainless Channel Letter Auto Profile Bending Machine Price, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Ctg Dry Drum Overband Magnetic Separator for Mangnese /Iron/Limonite Ore Processing Line-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Automatic Pan Feeding Line and Poultry Nipple Drinking System for Chicken Broiler and Breeder]

Aluminium / Stainless Channel Letter Auto Profile Bending Machine Price,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[Whole Set Gas Spring Strut for Furniture]

Aluminium / Stainless Channel Letter Auto Profile Bending Machine Price, 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:Factory Outlet Automatic Grain Tower Dryer Drying Equipment, Part I.Mesh Bag Packing 20kg Pure White Garlic. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Union Metal Steel Building and Construction Material]

Supreme 524 Mv Sea Kayak, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Trunk Organizer Foldable Water Proof.100%Polyester Burnout Velvet Sofa Fabric (pineapple grid), 175-184[Brass Material Air Vent Plug]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Factory Direct Sale Single Cloth Hanger Hook]

Whitaker,J.R.(1963),Diesel Generator Parts AVR Ki-Davr-95s for Kipor. Analytical Chemistry35(12), 1950-1953[27 Inch LCD Monitor with Capacitive Multi Touch Touch Screen for Computer Display]

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

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