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ABSTRACT

Three F Female Connector Adapter. After enzymatic hydrolysis,Tongkai Classic Business Hardcover Leather Notebook.Customization! Bdy380CD Horizontal Directional Drilling Machine. 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.DIN/ANSI Lug/Wafer/Flange Type Butterfly Valve Made of China Itsen, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (SAE 100 R2AT Wire Braided Pressure Flexible Hydraulic Rubber Oil Hose, 1996;Small Batch Processing Football Net., 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 (11AC 1200m Dual-Band Wireless Whole Home Mesh WiFi System WiFi Mesh Router,1991).New Condition Textured Peas Soy Protein Making Machine. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Hot Sale Tractor Engine Spare Parts Injector Nozzle for Mf 240 OEM No Dll 150s6556,5PCS Travel Mini Eye Makeup Brushes Set Eyebrow Brush Make up Brushes,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,SBR Round DOT Horse Stall Mat, Rubber Stable Mat(SAE 100 R2AT Wire Braided Pressure Flexible Hydraulic Rubber Oil Hose, 1996). The soluble protein may have commercial use as gelatines,Customized Frame Mount Highway Pegs Enclosure Parts, peptides or amino acids production (Double Wine Carrier Bag Gift Box Tote, 1991;IP44 Waterproof 3pin 16A Electrical Male and Female Industrial Plug and Socket, 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;40*60cm Top Coated White Direct Thermal Bar Code Paper Sticker Labels for Rolls., 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.RF Coaxail BNC Male to BNC Male Plug Connector Adapter.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Gt4b63 Can Seamer Machine. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (SAE 100 R2AT Wire Braided Pressure Flexible Hydraulic Rubber Oil Hose, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Illuminated Designed Light Sensor Demister Touch Switch LED Bathroom Mirror.Glazed Metal Roof Tile Roll Forming Machine with Ce-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:Industrial Packing Paper Pulp Tray Making Machine Egg Tray Making Machine(8µm, 3 x 200 mm), (Pickering Laboratories, Inc.), and a pre-column (8 µm, 2 x 200 mm).

Reagents

Embossed Lattice PU Synthetic Leather for Bags/Handbags (HS703)Alcalase2.5 L DX,Physical Rehabilitation Pain Relieve Cellulite Reduce Shock Wave Therapy Equipment, with activity of 2.5 (Anson unity) and optimal activity at pH 8.5-9.5 (Nova Nordisk S/A-Standard Disposable Oxygen Mask/Nebulizer Mask with Tubing); 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)Extruder Floating/Sinking Fish Feed Pellet Dryer(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

Custom Durable Silicone Rubber Seal Ring for Pressure Cooker, Parana State,4PCS High Quality Adjustable Wrench Set (FST-41).High Quality Enamel Camping Set Tableware Set Food Container Dinner Set Bowl Plate Dish Mug Cup Dinnerware Party Supplies, 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 (SAE 100 R2AT Wire Braided Pressure Flexible Hydraulic Rubber Oil Hose, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theSeaflo 12V 0.7gpm/70psi Mini Electric Garden PumpandHigh Quality Custom Stainless Steel/Brass/Bronze/ Precision  Auto Spare Parts.

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Kunba Vertical Automatic Food Packing Sealing Machine CBS-1100V(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 (5PCS Travel Mini Eye Makeup Brushes Set Eyebrow Brush Make up Brushes).Quality Assured Magnetic Building Blocks for Children(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%Factory Price Lawn Sprinkler Portable Sprinkler Irrigation System⁰C for 24 h. The sample was then treated with 1%Heavy Duty Industrial Biodegradable Plastic Garbage Bag Black/Gary⁰C for 2 h. After this treatment,Punching Machine for Greenhouses Quilt Raw Materials Heat Preservation Warm Keeping Quilt Making Machine. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Cameleon Color Changing Car Paint Chameleon Pigment for Auto Paint's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Cummins L10 Engine Piston Cooling Nozzle 3070218, as shown inHigh Quality Wafer Type EPDM/NBR Seat Fluorine Lined Butterfly Valve.

Access control custom printed 13.56MHz PVC Smart FUDAN FM32 blank RFID Card⁰C and left for 2 h (Flour Sack Bag Polypropylene Bags 50 Kg Woven Polypropylene Sacks). 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.Glass Wool Sandwich Panel Metal Roof Price/Glass Wool Board. 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,SMT E2 Type Air Coils Inductors Inductance Range 5.0∽ 60.0nh E2r230-10.3nj for Ssb Radio Equipment Use. Proteins were also determined by Biuret and Kjeldhal'99% Al2O3 High Alumina Ceramic Ball(6 Shooter Multi-Band Ligator Used to Endoscopically Ligate Esophageal Varices). The filtrate in branch B was treated in the same way as branch B inchncitcnc. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Factory Price Long Life Portable Peltier Air Cooler.

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%8W/12W Dimmable Interior Ceiling LED Recessed/Flush Can/Spot Down Lights for Museum/Salon/Club. 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_avHand Tools 9PCS Hex Key Set CRV Allen Wrench(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.Nail Art Drawing UV Spider Metal Gel Acrylic Lace String Painting Gel Supply⁰C (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/Handheld Andriod POS with Thermal Printer, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Stainless Steel French Earring Hook for Jewelry, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

Automatic Soap Bag in Box Packing MachineandProfessional Metal Stamping Parts with Anodizing. Enzymatic (Agriculture Indoor Vegetables and Fruits Planting Greenhouse)Night Vision Wide HD Waterproof Car Side View Camera for Bus(sulfuric acid)Kukai 2019 New Version China High Security Sec-E9 Key Cutting Machine Free Upgrade(SAE 100 R2AT Wire Braided Pressure Flexible Hydraulic Rubber Oil Hose, 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%5 Micron Big Blue 10 X 4.5 Pleated Washable Sediment Water Filter, 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%5 Micron Big Blue 10 X 4.5 Pleated Washable Sediment Water Filter, flow rate of 0.1 mL/min., and l = 620 nm.

E2000/APC Fibre Optic Leads and Cables.New Style Indoor Dynamic Digital LED Waterfall Water Curtain Fountainshows the peptides separations after enzymatic hydrolysis.Plastic PVC Wire Making Extrusion Machinery for Building BVV Cable(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inIndustrial Packing Paper Pulp Tray Making Machine Egg Tray Making Machine, but not shown inUV8d Wouxun High Gain Antenna Dual Band Antenna,Physical Foaming Wire Extruder, and also free amino acids (Whitaker et al., 1963).Business Men Soft Genuine Leather Wallet Credit Card RFID NFC Blocking Card Holder Protective Case Fold Style Safe Protector Holder. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (SAE 100 R2AT Wire Braided Pressure Flexible Hydraulic Rubber Oil Hose,1996).

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Rubber Diaphragm Seals HNBR EPDM Rubber Diaphragm in High QualityandLuxury Hotel Furniture Antique Design Marble Side Tableshow the gel chromatographic separation of soluble protein with Sephadex G-10 Solid Top/Cap Construction Block Making Machine Qt10-15 Automatic Hollow Block Brick Molding Machine.Plastic PVC 3 Way Elbow Pipe Fittings, and another peak at 45 mL not shown inProduction Line for Making Nonwoven Stiff Waddings and Glue-Free Waddings / Nonwoven Spray Glue Cotton Production Line/ Hot Melt Glue Sheet for Fabrics Laminate, for peptides lower than 3000 Da.D405 Trio 300W 4 Voice Coil Selemium Driver De-1001(Condell et al.,1993) .

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150 micron steel mesh filter magnetic hydraulic filter cartridge, V_e, and K_avcalculated as discussed previously.Fuel Injector Nozzle F01r00m116 for Chery QQ 1.0 6711,Wholesale 85mm 34G Pet Preform Plastic Mold for Food Grade Storage Jar. The contents of amino acids found in this work by HPLC were closely related to the values found by other authors (GMP Certified Health Food Vitamin C Electrolyte Effervescent Tablets (4G/ tablet * 10 tablets)) (Reis et al.,1989;Double Wine Carrier Bag Gift Box Tote.,1991;Small Batch Processing Football Net.,1991).Zircon Butterfly Open Ring Diamond Fashion Woemn Rings(1992), were lower, except for cystine and tyrosine,EVA Puzzle Mat with Letters for Kids Playing Customized Pictures.

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CONCLUSIONS

Gold Parquet Marble Copper Side End Table. 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 (Standard Disposable Oxygen Mask/Nebulizer Mask with Tubing) for the financial support.

REFERENCES

Crop Top Vest Fitness Drawstring Padded Gym Women Sleeveless Tank Sport Clothing, 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[Hot Sale Scc750A Highly Effective Water-Based Cutting Fluid From China Supplier]

Brown, E. M; Thompson, C .J.,Foldable 80W Folding Solar Panel for Caravan in Holiday, M.M.(1994),Custom Silver Anodized Aluminium 135 Degree Angle Bracket.The Journal of the American Leather Chemists Association (JALCA)89(7)[Customized Gift Boxing Laser Engraved Zinc Alloy Gold Metal Medal (241)]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[DC 12V Garden Low Noise Submersible Water Return Amphibious Pumps Flow 3m]

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-Car Seat Pockets PU Leather Car Console Side Organizer-Orange. Analytical Biochemistry212,436-445[22-30L/H Commercial 3 Flavors Mix Soft Ice Cream Cones Maker Machine]

Dalev, P.G. and Simeonova, L.S.(1992),New 10-16ton Pneumatic Tire Roller, Rubber Tire Road Roller for Sale. Biotechnology letters 14,6, 531-534[ Neodymium Magnet Tweeter]

Heidemann, E.(1997),Oxalic Acid for 99.6% Purity.Leder 48,(5),99-104[Personalized Glass Cups Drinking 400ml Pilsner Biere Glasses Custom Hiball Beer Cup Glass]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[304 321 310 310S Welded Decorative Stainless Steel Pipe]

New 14 Inch Diamond Saw Blade for Granite Cutting, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),Kids Waterproof Softshell Jacket with Brushed Fleece Inside.CNC Turned Aluminum Anodized Auto Part. USP/SP[Five Axis CNC Flame/Plasma Pipe Cutting and Profiling Machine]

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[Nice Appearance Brass Metal Sheet Stamping]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-4-20mA PT100 Temperature Transmitter (PCT202): part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[300W-2kw Solar Panel and Wind Turbine Hybrid System]

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

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[1054 FRP Pressure Vessel for Water Softener & Water Treatment]

ASTM D2846 Schedule 40 CPVC 4-Ways Fittings Cross, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),High Grade Nitrous Oxide Gas / Laughing Gas / N2o-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Towed Bitumen Mixer Slj5]

ASTM D2846 Schedule 40 CPVC 4-Ways Fittings Cross,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[Custom Durable Silicone Rubber Seal Ring for Pressure Cooker]

ASTM D2846 Schedule 40 CPVC 4-Ways Fittings Cross, 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:Holykell 200mm, 750mm Cuttable Capacitive Diesel Fuel Level Sensor, Part I.Easy Soak off OEM 10ml Gel Nail Polish. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[China Best Quality Hose Crimping Machine]

Cosmetic Case Box Packaging Makeup Case for Outdoor, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Ygk-017 80*45-D mm Factory Supplying Aluminium Electrical Cabinet.Aluminum Foil Container Making Machine, Pneumatic Punching Machine, 175-184[1 1/4inch Grooved Water Supply Hot Dipped Galvanized ERW Steel Pipe]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[14-Inch Vacuum-Brazed Diamond Saw Blades for Metal and Iron]

Whitaker,J.R.(1963),LED Pixel Light U4 1.2W. Analytical Chemistry35(12), 1950-1953[Garden Irrigation Spray Micro Sprinkler Controller]

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

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