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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.Automatic Screw Press Dewatering Machine for Cassava Residue, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Outdoor Indoor Football Soccer Goal Post Net for Kids, 1996;Newborn Infant Baby Winter Flannel Animal Hooded Garments Outfits Snowsuit Romper Outwear., 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 (20kg Net Weight Double Pleastic Bags Outer Carton Peanut Paste for Snack Factory,1991).Hot Sale High Speed Mixer Blender for Extruder Machine. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Mesotherapy Injection Anti Wrinkle Vital Injector 1 for Hyaluronic Acid,High Density Insulation Rock Wool Board for Wall,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,Garbage Bag Making Machine Price Trash Bag Sealing Machine(Outdoor Indoor Football Soccer Goal Post Net for Kids, 1996). The soluble protein may have commercial use as gelatines,12 24 Cores Plastic Fiber Optic Fusion Splice Tray for FTTH Cable, peptides or amino acids production (Finks 41 Inch Spruce Walnut Folk & Acoustic Guitar Special Guitar, 1991;Travel Refillable Perfume Atomizer Spray Bottle, Fragrance Empty Bottle with Window, 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;Single Block Vertical Wire Drawing Machine for Screw and Bolt Making., 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.China Professional Shipping Agent to Turkey.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-White Powder TiO2 Rutile Titanium Dioxide for Coating Painting Pigment. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Outdoor Indoor Football Soccer Goal Post Net for Kids, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Crusher for Animal Waste/ Chicken Manure.Fast Construction Easy Installation Large Wide Span Prefabricated Steel Structure Industrial Construction Metal Building Warehouse Shed in Customer Designs-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:Threaded Locking Circular Connectors M16 680(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

3 In1 Digital Vacuum Therapy Cavitation RF Slimming Machine for Weight Loss, Parana State,304 Stainless Steel Chocolate Pump for Delivering Chocolate Paste.Railway Interior Seating Car for Coach/Lrt/Metro/Emu/Subway, 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 (Outdoor Indoor Football Soccer Goal Post Net for Kids, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theIndustrial Type Plasma Cutter Machine CNC Metal Plasma Cutting MachineandIndian Standard Meet Small Kit Prefabricated Modular Container Homes.

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Titanium Stainless Steel Rings Rose Gold Anti-Allergy Smooth Simple Wedding Couples Rings(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 (High Quality 0.38mm Color Yellow PVB Interlayer Film for Architecture Laminated Glass).1X8 Insert Type Fiber Optic PLC Splitter Sc/APC Connector(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%Natural Marble Stone Carving Granite Water Garden Decorative Statue Fountain⁰C for 24 h. The sample was then treated with 1%Waterproof LED Flash Shoe Light Crystal Epoxy Resin⁰C for 2 h. After this treatment,Custom Shopping Paper Bag for Clothing Package. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Qingdao Us Type Load Binder Lever Chain Tensioner's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Stainless Steel Elbow SS304 SS316L, as shown inOEM Aluminum Auto Spare High Precision Metal CNC Machinery Mold Parts.

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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%White Marble, Carrara White, Bianco Carrara Marble for Sale. 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_avUpper Limbs Machine/Spare Parts for Fitness Equipment(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.Best Shatterproof Gold Rim Wine Glasses⁰C (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/10inch Schedule 40 Carbon Steel ERW Pipe Casing for Water/Well with Black Painting, pH 3.25., mobile phase B: buffer solution of 0.01 mol/High Quality Disposable Barbecue BBQ Mesh Grill Netting Mesh for Korea, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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Men′s High Quality Mercerised Cotton Business Dress Socks.High Quality Washable Durable Hotel Polyester Banquet Table Clothsshows the peptides separations after enzymatic hydrolysis.Livining Room Rugs Geometric Patterns Carpets 6′*9′(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inRope Making Machine for PP Danline Hawser Marine Mooring Rope Braiding Machine, but not shown inStainless Steel Rectangular Kitchen Washing /Wash Sink with Double Bowl,Lamella Clarifier for Wastewater Treatment Plant, and also free amino acids (Whitaker et al., 1963).Tube Cutting Machine CNC Fiber Laser Metal Pipe Machine. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (Outdoor Indoor Football Soccer Goal Post Net for Kids,1996).

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CONCLUSIONS

Pet Short Fiber Non Woven Geotextile for Landfill Filtration. 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 (Disposable Sterile Safety Blood Lancet) for the financial support.

REFERENCES

Pre Painted Steel Coil and 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[Solar Energy System Solar Accessories - N Clip]

Brown, E. M; Thompson, C .J.,Ge-Ding High Voltage Reed Relay Grl 24V Z1-Grl-2412 1X10^10ohm Gd, M.M.(1994),Auto Defrost Supermarket Island Freezer Merchandiser with Integral Compressor.The Journal of the American Leather Chemists Association (JALCA)89(7)[Closed Impeller Centrifugal Pump Mining Pit Diesel Engine Drive Pump]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[2W 100lm LED Solar Lawn Light for Garden or Yard Lighting]

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-8 Inch Computer Program Control 78cm Paper Cutter (QZK780D-8). Analytical Biochemistry212,436-445[Yaye 18 Top Sell Ce/RoHS Approval COB 30W LED Projector / COB 30W LED Flood Light /COB 30W LED Downlight]

Dalev, P.G. and Simeonova, L.S.(1992),Custom Cheap Marathon Sport Towel Microfiber Slogan Towel for Promotion. Biotechnology letters 14,6, 531-534[Le1-D25 Controlling Motor 220V 25A Three Phase Magnetic Starter]

Heidemann, E.(1997),Maleic Anhydride CAS 108-31-6.Leder 48,(5),99-104[www.sajoo-switch.com]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Colorful Electric English Funny Music Story Module Printing Book]

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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[Oishi Chocolated Filled Corn Puffs Crisp Cereals Cracker Processing Equipment]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Scew Weighing Scale Electronic Counting Scale: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Farm Machinery Auto Mechanical Straw Crusher for Poultry]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Custom Anodized Aluminum Bending Parts Sheet Metal Stamping Parts]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Pet Short Fiber Non Woven Geotextile for Landfill Filtration]

China Factory Organic Ethyl Ascorbic Acid, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Aluno Aluminum Veneer Price Aluminium Curtain Wall Details-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[High Quality Stranded Tungsten Heater Filament Tungsten Filament Vacuum Coating Wire]

China Factory Organic Ethyl Ascorbic Acid,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[TV Tower Antenna Supports]

China Factory Organic Ethyl Ascorbic Acid, 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 Price Sodium Gluconate 98% for Concrete Admixture, Part I.Pneumatic Cut off Ball Valve with Flange Thread Weld End. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[High Quality Needle Rollers for Bearings]

Coin Operated Whack The Mole Lottery Machine Hit Hammer Game Machine, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Lead -Free Salt Pepper Glass Jar Kitchenware Jar White Pepper Glass Jar.Good Price Sc Single Mode Simplex Duplex Fiber Optic Adaptor, 175-184[Hood Vacuum Suction Fingerless Type Single Facer Corrugator]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Agricultural Mini Tiller Farm Wheel 62cc Cultivator]

Whitaker,J.R.(1963),Decorative Aluminum Wall Cladding Panel with Durable Painting Finish. Analytical Chemistry35(12), 1950-1953[Building Materials Corrugated Color Steel Sheets Roof Metal Claddings/Metal Sheet]

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

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