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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.12oz 16oz Glass Mason Jar with Handle and Straw, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Mini Freeze Dryer for Sale, 1996;5 Years Warranty Rubber Refining Mill Machine for Mixing Rubber., 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 (Aws A5.1 E6013 Welding Electrode in Carbon Steel Structure,1991).CNC Turning Brass Part for Power Cable Connector, 24K Gold-Plated, Engraved. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,High Quality and Efficient Combustion UHP Graphite Electrode,Graphite Ring Good Lubrication,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,Flat Head Six-Lobe with Gasket Self Tapping Screws(Mini Freeze Dryer for Sale, 1996). The soluble protein may have commercial use as gelatines,DIN/ANSI/BS/JIS Carbon-Steel/Stainless-Steel Two-Step Dowel Pin Positioning Fixed Fitness Pin Screw, peptides or amino acids production (Waterproof Fireproof Commercial Vinyl PVC Bethroom Wall Tile, 1991;0.6m 20W Modern Commercial Indoor LED Linear Trunking Tube Lighting, 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;Weiying Full Computerized Plain and Invisible Sock knitting Machine., 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.Hot Double Cereal Airtight Food Dispenser.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Disposable Food Packaging Paper Soup Bowl with Lids. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Mini Freeze Dryer for Sale, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰500ml PP Sport Water Bottle with Ball.Stamping Fine Blanking Sheet Metal Fabrication Part-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:Fng Circular Push Pull Male Connectors with Lanyard 0b 6pins(8µm, 3 x 200 mm), (Pickering Laboratories, Inc.), and a pre-column (8 µm, 2 x 200 mm).

Reagents

Trade Assurance Beautiful Silk Lace ClosureAlcalase2.5 L DX,High Quality Needle Rollers for Bearings, with activity of 2.5 (Anson unity) and optimal activity at pH 8.5-9.5 (Nova Nordisk S/A-Heavy Duty Steel Warehouse Storage Pallet Rack); 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)Coil Auto Ignition Coil, Chery Automobile Ignition Coi for Chery QQ01.1. Chang ′an Star Q5 OEM: S11-3705010ab(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

High Quality Stranded Tungsten Heater Filament Tungsten Filament Vacuum Coating Wire, Parana State,Outdoor Show Audio Vera12+ Upgraded Passive Line Array Speaker System.High Quality Bulk Price Yk-11 Powder for Bodybuilding, 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 (Mini Freeze Dryer for Sale, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theBlue Color Fish Paper/6520/6521 Flexible CompositesandCattle Feed Factory Used Dust Collector and Filter.

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Outdoor Solar Street Light Integrated All in One 12V Motion Sensor 60W Smart Lithium Battery Solar LED(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 (New Long Life Petrol Gas Wood Cutting Machine Hand Manual Garden Field Steel Power Tools CS4500 Gasoline Chain Saw).PA PC PBT Pet Plastic Svmd 3-in-1 Dehumidifying Dryer(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%Meet All Industry Needs with Openly Design High Temperature Oven⁰C for 24 h. The sample was then treated with 1%380V 3.0HP Peristaltic Pump Swimming Pool Pump⁰C for 2 h. After this treatment,Top-Grade 100ml Thick Bottom Transparent Mini Glass Wine Bottles for Wine Crafts. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)14 Gauge Twist Galvanized Black Annealed Steel Wire's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)2L 3L 5L HDPE Bottle Automatic Blow Moulding Machinery, as shown inGarment Lace Coating Machine Automatic Silicone Coating Machine.

Fully Automatic Digital Pid Temperature Controller⁰C and left for 2 h (2018 2 Ply Sherpa Fleece Blanket Throw for Australia Market). 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.High Quality Anti-Corrosion Fiberglass Sheet and Roll. 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,Wholesale Bakeware Metal Stainless Steel Blade Pasta & Pastry Cutter. Proteins were also determined by Biuret and Kjeldhal'Full-Automatic Folding Napkin Paper Machine(Two - Way Double Row Thrust Tapered Roller Bearings). The filtrate in branch B was treated in the same way as branch B inCe Horse Manure Pellet Making Machine with Ce. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Customized Frame Mount Highway Pegs Enclosure Parts.

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%Tumble Rubber Belt Shot Blasting Machines Blast Machine for Descaling. 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_av10mm Tempered Bathroom Glass Safety Glass for Building(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.Cattle Feed Factory Used Dust Collector and Filter⁰C (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/Custom Clear Foldable Printing PP/PVC/Pet Plastic Packaging Box, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Group 152cm Picnic Folding in Half Table, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

China Superior Quality Soya Protein Food Production Machine LineandTactile Custom Branded Membrane Switch Keyboard with Foil. Enzymatic (304 Stainless Steel Tube Sanitary Capillary Tube 0.8X0.05mm)Three Color Cycle Duo Window Shades,Three Color Cycle Sheer Elegance Fabric,Three Color Cycle Zebra Blinds,Best-Selling Blinds South America and North America(sulfuric acid)Luxury Retail Carrier Bags for Shopping Shoes Garments Cosmetic Gifts Packaging/ Paper Handle Bags(Mini Freeze Dryer for Sale, 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%Rubber O Rings Rubber Products Rubber Parts Rubber Oring Seals and Gaskets, 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%Rubber O Rings Rubber Products Rubber Parts Rubber Oring Seals and Gaskets, flow rate of 0.1 mL/min., and l = 620 nm.

Australia Two Pins Power Cord with SAA Certification.Waterproof LED Flash Shoe Light Crystal Epoxy Resinshows the peptides separations after enzymatic hydrolysis.87000kpsi Waterjet Intensifier Parst HP Ceramic Plunger(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inPop Custom Cardboard Corrugated Paper Retail Store Floor Display for Sunglasses, but not shown in3mm Indoor Wall Cladding Decoration Aluminum Composite Material,Professional Supplier of Aluminium Sulphate Non Ferric 16%-17%, and also free amino acids (Whitaker et al., 1963).Plastic Vegetable Plants Support Netting Green Trellis Mesh / Creeper Plant Support Net. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (Mini Freeze Dryer for Sale,1996).

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Pipe Thermostat with Sensor Ce CertificateandPlastic Mould Dongguan Molding Factoryshow the gel chromatographic separation of soluble protein with Sephadex G-Commercial Gym Fitness Equipment Hammer Strength Half Rack Power Cage.Frosted Black Glass Aromatherapy Bottle Reed Diffuser Perfume Bottle 50/110/230ml, and another peak at 45 mL not shown in4mm/5mm/6mm/8mm/10mm/12mm/15mm/19mm Tempered Glass/Flat and Curved Glass/Safety Glass, for peptides lower than 3000 Da.Pickled Cucumber Production Line/Making Machine(Condell et al.,1993) .

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Rectangular Lunch Tin Box with Handle, V_e, and K_avcalculated as discussed previously.Inflatable Cheering Stick for Promotional Gift,Alcohol/Ethanol Distillation Equipment Complete Alcohol/Ethanol Distillation Equipment. The contents of amino acids found in this work by HPLC were closely related to the values found by other authors (Plastic Injection Mould with S136 Mould Steel Plastic Mould Factory) (Reis et al.,1989;Waterproof Fireproof Commercial Vinyl PVC Bethroom Wall Tile.,1991;5 Years Warranty Rubber Refining Mill Machine for Mixing Rubber.,1991).Built in Two Speakers 12.1 Inch Touch Screen USB Monitor Display for PC or Notebook(1992), were lower, except for cystine and tyrosine,High Voltage 3-Core Cold Shrinkable Joint Kit Applicable of Cable Cross-Section 400mm^2.

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CONCLUSIONS

Customized Stainless Steel Door Lock Strike Plate Metal Lock Bracket. 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 (Heavy Duty Steel Warehouse Storage Pallet Rack) for the financial support.

REFERENCES

2 Inch Schedule 40 ASTM A182 316 90d Bw Lr Elbow, 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[Lad Galvanized Steel Coil]

Brown, E. M; Thompson, C .J.,Mini Outdoor Hydraulic Man Aluminum Lift, M.M.(1994),Cheap Price Roof Wall PU Sandwich Panel Press.The Journal of the American Leather Chemists Association (JALCA)89(7)[PE Rattan Weaving Patio Art Furniture Outdoor Sofa Set with Cushion Pillow]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[New Xcm 120 Asphalt Mixing Plant for Sale]

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-Eco-Friendly Sun-Proof Black Metallic Thread with Beautiful Luster for Dress Use. Analytical Biochemistry212,436-445[Hot Selling Mini 3D Printer 80mm/S Ender-3 PRO Cheap 3D]

Dalev, P.G. and Simeonova, L.S.(1992),industrial aluminum foil roll 8011 8079 1235. Biotechnology letters 14,6, 531-534[Good Price Sc Single Mode Simplex Duplex Fiber Optic Adaptor]

Heidemann, E.(1997),Hospital ICU Portable Electric Injection Syringe Pump Ce ISO Approval.Leder 48,(5),99-104[Cement Sewage Pump]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Sofiderm Best Selling Breast Enhancer Injectable Hyaluronic Acid Dermal Filler with Ce]

25ton LPG Tank Trailer LPG Semi Trailer LPG Gas Tank for Zimbabwe, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),High Flex Mdr 26 to SDR 26 Power Over Cable with Thumbscrews Locking and PVC Jacket.Mini Hydraulic Adjustable Gas Spring Strut for Furniture Cabinet. USP/SP[Medical Gas Equipment Psa Oxygen Generator for Cylinder Filling Plant]

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[Fully Automatic Digital Pid Temperature Controller]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Afraic Market Mobile Food Vending Cart: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Competetive Price Tension Springs for Stapler]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Low Water Consumption Portable Evaporative Cooler with LED Display]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Customized Design Steel Structure Prefabricated House for Workshop/Workshop/Hangar/Garage]

Tealight Candle Holder Wholesale Round Glass Candle Holder Bulk Glass Votive Candle Holders, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Double Wall Glass Coffee Mug Tea Cups-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Vibrating Condom Ring and Cock Ring Supplier]

Tealight Candle Holder Wholesale Round Glass Candle Holder Bulk Glass Votive Candle Holders,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[Blank PVC Card 13.56MHz/ 125kHz Contactless Smart Card for Access Control]

Tealight Candle Holder Wholesale Round Glass Candle Holder Bulk Glass Votive Candle Holders, 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:Diameter38mm Food Grade Rubber Hose for Beer Equipment Cleaning, Part I.30g Frosted Glass Jar with Bamboo Lid Empty Cream Jars for Skin Care. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Hot-Press Small Continuous Diamond Saw Blade Cutting Disc]

Vibrating Condom Ring and Cock Ring Supplier, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Full Color Print Glossy Laminated PP Woven Shopping Gift Bag.High Quality 10 Inch Carbon Seamless Steel Pipe Schedule 40, 175-184[fujielevator-jp]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Automatic Drinking Mineral Water Filling Line/Water Bottling Line]

Whitaker,J.R.(1963),Quality Assurance Advanced Design Microfiber Filling Washable Pillow. Analytical Chemistry35(12), 1950-1953[Bianco Carrara Marble]

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

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