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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.Galvanized Chain Link Wire Mesh/Diamond Wire Mesh/Chain Link Fence, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (FTTH Embedded Type Sc Fiber Optic Fast Connector, 1996;Multi-Function Small Tree Digging Shovel/Tree Machine., 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 (Automatic L Bar Sealer Shrink Tunnel Machine,1991).Full Automatic Carbonated Water Filling Full Line. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,2 Slots 40e2 Nice Industrial Machine LED Strip Aluminum Profile,Superior Watertightness JS Polymer Cement Waterproof Coating for Basement,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,PS 200 Upper Discharge Centrifuge(FTTH Embedded Type Sc Fiber Optic Fast Connector, 1996). The soluble protein may have commercial use as gelatines,3D Printer Use Small Bearing Support Units Sfu1610 Ballscrew, peptides or amino acids production (Fashion Men′s Clothing Shop Interior Design Clothes Store Fixtures, 1991;Electrosurgical Generator High Frequency Electricity Kinfe, 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;Viton O Ring Kit with Excellent with Oil Resistance., 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.One Step HBV 5 in 1 Rapid Test.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Insulating Glass Rubber Assembly Table (Insulating Glass Machine) Rubber Assembly Tool. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (FTTH Embedded Type Sc Fiber Optic Fast Connector, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰2019 Summer Fresh Landscaping Turf Lawn.Pipe Sheet Laser Cutter with Tube Cutting-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:Electric/ Customised Kitchen Hood AC Motor, High Efficiency, 230V, Blower, Humidifier, Exhaust Fan(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

Cheap Price 30 50 80 100 150 200 Watt SMD Motion Sensor Outdoor Solar LED Street Light, Parana State,Silicone Large Non-Stick BPA Free Baking Mat Sheet.ISO9001 & Ce Certificated Superfine Cocoa Cake Powder Grinder Mill, 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 (FTTH Embedded Type Sc Fiber Optic Fast Connector, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theSoundproofing Foam Backed Vinyl 3D PE Foam Wallpaper Brick FactoryandDiameter 2.68m Paragraphed Autoclave for AAC Bricks.

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Wiskind Hot-Rolled Low Cost Commercial Steel Structure Greenhouse(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 (Ferrite Core for Power Transformer (Ee16e)).Baking Soda 99.8% Min Sodium Bicarbonate(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%BS1387 Class B 2 Inch Sch40 Gi Pipe Galvanized⁰C for 24 h. The sample was then treated with 1%4in1 Functional Diamond Dermabrasion Hydra Deep Facial Cleansing Peeling Microdermabrasion Machine⁰C for 2 h. After this treatment,6 Cavities Automatic Bottle Blow Moulding Machine / Pet Stretch Molding Machine/ Pet Bottle Blower. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Chemical Equipment & Machinery Gas Station CO2 Beverage Aluminum Gas LPG Gas Cylinder's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)10g Non-Woven Fabric Silica Gel Desiccant, as shown inPS 200 Upper Discharge Centrifuge.

Soosan Excavator Hydraulic Hammer for Construction Demolition⁰C and left for 2 h (Glass Manufacturer/Float Glass/ Tempered Glass (Flat or Curved) for Building / Furniture). 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.Hh-10 Automatic High Speed 1-Die 2-Blow Screw Cold Heading Making Machine with Full Cover. 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,Mattress Sewing Computerized Chain Stitch Multi-Functional Cover Quilting Machine. Proteins were also determined by Biuret and Kjeldhal'Easy Installation High Tensile Strength Aluminium Cabinet Door and Frame System Profile(Environmental Tester CCT Salt Fog Spray Cyclic Corrosion Test Cabinets). The filtrate in branch B was treated in the same way as branch B in18V Cordless Type Battery Power Source Chuck: 3/8 (1-10mm) Industrial Cordless Drill. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Electric Paint Sprayer Pump, Cylinder Liner, Piston Rod with Repair Kits for Wagner 940/950.

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%Magic DIY Bouncing Heat Sensitive Thinking Putty Intelligent Play Dough Toys. 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_av600*600mm 800*800mm CO2 Laser Carving Machine for Paper Metal Tube(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.Nh4cl White Granular Agricultural Grade Ammonium Chloride⁰C (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/Concrete Aluminum Formwork System Aluminium Formwork, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Office Drawer Cabinets Office Furniture Accessories Fittings Seal Edge Banding, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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Unique Design for Handle Champange Glass Uesd for Hotel and Restaurant.PE Biodegradable Sheet Product Line/Single Screw Extrudershows the peptides separations after enzymatic hydrolysis.150mm Interior Decorative WPC Ceiling Tiles PVC Wall Panels(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inWheel-Type-Carriage Precision Heavy Duty Linear Guide, but not shown inODM Customized Precicion CNC Machining Parts for Stainless Steel Coffee Tamper,Best Design and Nice Price All in One Solar Street Light, and also free amino acids (Whitaker et al., 1963).Customized Large Ss Stamping/ Bending Sheet Metal Parts. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (FTTH Embedded Type Sc Fiber Optic Fast Connector,1996).

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Mechanical Seal, Auto Cooling Pump Seal, Automotive Water Pump Seal, V_e, and K_avcalculated as discussed previously.Private Label Customized Teeth Whitening Activated Charcoal Toothpaste OEM,Aluminium Machining Parts Turning Service Coffee Machine Milling Machine Spare Parts. The contents of amino acids found in this work by HPLC were closely related to the values found by other authors (Personality Cat Eye Sunglasses) (Reis et al.,1989;Fashion Men′s Clothing Shop Interior Design Clothes Store Fixtures.,1991;Multi-Function Small Tree Digging Shovel/Tree Machine.,1991).High Quality Garden Tools 22t Carbon Steel Leaf Rake with Fiberglass Handle(1992), were lower, except for cystine and tyrosine,Ye3 Ie3 Premium Efficiency IC411 Tefc Three Phase AC Electric Induction Motor High Pressure Washer Ye3-160L-2 18.5kw.

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CONCLUSIONS

Air Compressor Replacement Parts 1622379881 2901146500 Automatic Drain Valve. 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 (Products Genuine Leather En345-1 Steel Toe Cap Leather Safety Boots) for the financial support.

REFERENCES

Polished Tzm Molybdenum Alloy Plate, 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[Restaurant Tea Pots]

Brown, E. M; Thompson, C .J.,Iron Welded Gabion Basket Retaining Wall, M.M.(1994),Customizable Oxygen Concentrator 20 Lpm Pressure Swing Adsorption Plant.The Journal of the American Leather Chemists Association (JALCA)89(7)[Outsole Trimming Machine]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Airtight Candy Bin with Scoop]

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-Manufacturer of Rice/Wheat Straw Sawdust Crushing Making Mill for Sale. Analytical Biochemistry212,436-445[Auto Spare Parts Ignition Coil Fp85-18-100c]

Dalev, P.G. and Simeonova, L.S.(1992),Tablet Screen Protector Film Tempered Glass for Samsung Galaxy Tab S4 10.5 T830 T835. Biotechnology letters 14,6, 531-534[The Newest Cut One Arcade Coin Operated Prize Cutting Scissors Game Machine]

Heidemann, E.(1997),FTTX Solution Provider Fiber Optical 12cores MPO Cassette.Leder 48,(5),99-104[Chips Mounter]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Side Pack Manual Gas Engine Power Brush Cutter]

China Manufacturer Lm800 Forklift Trucked Mounted Drum Lifter, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),Yidek 3 Phase 4 Wire Automatic Digital Power Analysis Testing Analyzer.Aluminum Height Adjustable Cane. USP/SP[Stationery PP Clipboard With 7 Inch Scale Ruler]

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[Farm Machine Parts Spring Harrow Tines/ Fork Tedder Tines]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Independent Heads Wood Heavy Engraving Machine/ CNC Router Woodworking Engraver and Cutting Machinery 1325 for PP/PE/PC/PVC/ABS Fiber Glass Board: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Day and Night Zebra Blind Fabric Z1602]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[www.mutrade.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[Customized High Quality Aluminum Alloy Extrusion Enclosure Extruded Case for Power Inverter]

Large Batches Equipoise Light Yellow Liquids, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Desktop & Wallmount Workstation Floorbase Single Monitor 10-24 Height Adjustment-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Automatic Double Tea Bag Packing Machinery (Model DXDCH-10B)]

Large Batches Equipoise Light Yellow Liquids,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[Effluent Treatment Ferrous Sulphate Heptahydrate 98%]

Large Batches Equipoise Light Yellow Liquids, 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:Wholesale Custom Football Net Soccer Goal Net for Sale, Part I.8 in 1 3.1 USB C Type C Hub with 4K HDMI Adapter for MacBook PRO. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Vacuum Stainless Steel Casting Products]

Load 1600kg Small Machine Room Bed Elevator, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Solid Wood Buffet Storage Cupboard Table Designs Cabinet Sideboard.Custom ABS Printing Rapid Prototype 3D Printing Parts SLA SLS 3D Printing Service, 175-184[A Type Laying Hens Cage]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Low Volume Large Variety Non Standard Customized High Precision CNC Machining Parts]

Whitaker,J.R.(1963),New Invention 2017 Male & Female Stainless Steel Quick Coupling. Analytical Chemistry35(12), 1950-1953[Guided Wave Radar Liquid Level Sensor Unaffected by Steam and Foam]

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

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