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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.100ml/120ml/125ml Glass Bottle for Whisky/ Brandy/Rum/Wine, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (New Styles Fashion Erotic Women Underwear Sexy Lingerie, 1996;High Quality 1530 Metal Cutting Cheap CNC Plasma Pipe Cutting Machine with Rotary., 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 (Custom Precision Sheet Metal Forming Bending Welding Parts Housing Stamping,1991).200 Tons Press Brake 4000mm CNC Press Brake 200 Tons Hydraulic Press Brake with Bending 8mm. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Fully Automatic Water Bottle Lids Labeling Machine,2u Lgx Swing out Fiber Optic Patch Panel with 4 SC Simplex Single Mode Adapter Panels,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,New CCTV 4.0MP H. 265+ IR Bullet Waterproof IP Camera(New Styles Fashion Erotic Women Underwear Sexy Lingerie, 1996). The soluble protein may have commercial use as gelatines,Non-Woven Anti-Static Briefs...Non-Woven Anti Static Briefs, peptides or amino acids production (Fused Silica Quartz Lump 5-50mm for Refractory Materials, 1991;Cold Drawn D2 H13 Alloy Steel Turning Roll, 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;Interlocking Anti-Fatigue Kitchen Rubber Mat, Kitchen Floor Mat., 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.Bread Box Bamboo Large Capacity Roll Top Bread Holder Cupcake Storage Food Bin for Loaf Pastry Snacks.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-High Quality Suitcase Frame or Profile Bending Machine with Ce Aprroved. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (New Styles Fashion Erotic Women Underwear Sexy Lingerie, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Colorful 100LED Copper String Light Outdoor LED.H365 Emas Chainsaw Spares Cylinder Kits-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:PP Woven Bag for Feed, Rice, Corn Feed Bag Rice Bag 10, 25, 50 Kg(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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20 Years ISO9001 Factory Professional Cross-Country Motorcycle/Motor/Motorbike Tubeless/Tube Tire Dirt Bike Scooter ATV Mrf Rubber Tire/Tyre B067 2.75-21(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 (Natural Birch F&B Veneer Furniture Plywood).8 Pieces Hot Sale OEM&ODM Manufacturer Cruelty-Free Powder Blush Eye Cosmetics Makeup Brush(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%Hybrid Inverter Combined MPPT Controller 1-12kw⁰C for 24 h. The sample was then treated with 1%QC11y-10X3200 Hydraulic Guillotine Shearing Machinery/Steel Plate Cutting Machinery⁰C for 2 h. After this treatment,Genuine Original OEM Kubota/Isuzu/Yanmar/Cummins/Foton/Yto/Omro Products Spare Parts. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Newest Men Watches Popular Products Mens Custom Wood Watch's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Accurate Measuring Rulers 3D Lenticular Plastic Rulers, as shown in800mm Length 600n Load Tension Gas Spring with Metal Eyelet.

Reusable PLA and Pbat Tea Packing Pouch Supplier in China⁰C and left for 2 h (Strong Electric Locksmith Lock Pick Gun Tools for Bump Door Lock Opener). 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.Dental YAG Laser Welding Machine for Gold Silver Metal Jewelry. 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,Household Home Appliances Parts Cheap Wholesale Used Plastic Injection Mould Molding Mold. Proteins were also determined by Biuret and Kjeldhal'Industrial P-Toluene Sulfonic Acid for 93%-99% Purity(Tungsten Solid Carbide Straight 4/6 Flutes Reamer). The filtrate in branch B was treated in the same way as branch B inDlsf Series Filter Cartridge for Solvents Filtration in Biopharm Market. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Longkou Fushi Automatic PS Foam Thermocol Container Forming Machine.

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%Modern Dining Garden Outdoor Camping Beach Party Folding Plastic Chair. 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 Purity99.95% Cobalt Oxide(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.Leather XPE 5D Car Floor /Trunk/ Cargo Mat for Cadillac Car⁰C (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/Best-Quality Plywood Automatic CNC Hydraulic 800 T Hot Press Machine, pH 3.25., mobile phase B: buffer solution of 0.01 mol/8.8 Grade DIN931 DIN933 DIN934 Black Steel Hex Bolts, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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Tomato Paste Making Machine-Turn Key Solution.CNC Metal Grinding Machine Tool Mk2120shows the peptides separations after enzymatic hydrolysis.Vegetable Cutter Kitchen Gadget 330ml Mini Size Handy Pull Chopper, String Pulling Food Slicer and Chopper(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared in High Strength FRP/GRP Square Pipe Tube Gh F001 Guohao021 FRP Product, but not shown in6090 CO2 Sealed Glass Cooling Laser Cutting Machine,Automatic Four Colors Label Screen Printing Machine UV PCB PVC Printing Machine, and also free amino acids (Whitaker et al., 1963).HPMC Similiar to Tylose Powder for Rapid-Set Adhesive. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (New Styles Fashion Erotic Women Underwear Sexy Lingerie,1996).

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CONCLUSIONS

Samsung LTI550HN16 1920X1080P monitor screen LCD video wall. 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 (Adjustable UV Protection Brim Plain Snapback Transparent Baseball Cap) for the financial support.

REFERENCES

Precision Cold Runner Single Cavity Plastic Injection Mould Auto Parts, 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[Best Prices Prestressed Spun Reinforced Concrete Electric Pole Making Machines]

Brown, E. M; Thompson, C .J.,Household Family Medical Emergency Electronic Digital Thermometer, M.M.(1994),Die Cast Iron Worm Gear Speed Reduce Gearbox.The Journal of the American Leather Chemists Association (JALCA)89(7)[Automatic Thermoforming Making Machine for Plastic Egg/Battery/Fruit/Cake/Fast-Food Tray Container]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[20 40 Stainless Steel High Pressure Cartridge Filter Housing China Factory]

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-19*15W RGBW Beam DJ Disco Stage Light LED Moving Head Lighting. Analytical Biochemistry212,436-445[Wood Chips Rotary Dryer Sawdust Pellet Drying Machine]

Dalev, P.G. and Simeonova, L.S.(1992),High Quality Couple Watches Steel Material Gift Watch OEM in China. Biotechnology letters 14,6, 531-534[Korea Hot Sale 24K Goldzan Ampoule 99.9% Pure Gold Face Essence]

Heidemann, E.(1997),Glass Tea Sugar Coffee Storage Jar with Screw Cap.Leder 48,(5),99-104[Orfs Female Flat Seat]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[High Quality Beech Wood Chiavari Chair at a Competitive]

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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[Glass Tea Sugar Coffee Storage Jar with Screw Cap]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Particle Board Manufacturer, Melamine/Plain Chipboard: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Teflon Seal/Seals Spring Energized]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[m.agapassworld.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[Metal Hand Folding Machine 1.5mm Aluminum Sheet Manual Bending Machinery]

CNC Milling Machinery Parts From OEM Manufacturer/ Auto Part, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Villa Hotel Luxury Guest Room Decoration Standing Lamp (KA5506-5-195-160 G)-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Stainless Steel Bend Pipe for Industrial Use]

CNC Milling Machinery Parts From OEM Manufacturer/ Auto Part,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[Ss Wafer Type Check Valve Dn32 Dn40 Dn50]

CNC Milling Machinery Parts From OEM Manufacturer/ Auto Part, 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:Ttp60fras Metal Hard Alloy Tool Carbide Inserts Cutting Hand Tool Machine Tool End Tool Drill Milling Cutter Tungsten Carbide Cutter Carbide Insert, Part I.Outdoor HD P5.95 Mobile LED Display Panels with Epistar Lamps. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[New Design High Quality Alminum Sandwich Panel High Pressure Laminate]

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Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[99% Purity Serms Steroids Megestrol Acetate CAS 595-33-5]

Whitaker,J.R.(1963),Gc3 Lead Screw Ball Screw with Double Nuts 6000mm Ball Screw for Overhead Crane. Analytical Chemistry35(12), 1950-1953[Durable TPR Pet Molar Toy with Colorful Rope]

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

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