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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.Eco-Friendly Round Head Screw Stud Flange Hexagon Bolt for Building, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Polypropylene Woven 25kg 50kg Bags Wholesale Sand Bags, 1996;2-25mm Co-Extruded Highlight PVC Foam Board., 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 (CNC Engraving Machine Laser Marking for Assembly Line,1991).PP/PE Plastic Lawn Turf Gravel Grass Grid for Driveway Paver (PGG5050A-1). Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Personalized Transparent Waterproof Plastic Bag Wet Underwear Clear PVC/EVA Ziplock Packing Bag with Gusset for Swimwear,High Quality A3 High Speed Epson Head UV Printing Machine for Bottle Printing,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,12V/24V, Ts16949, Air Conditioner Fan for Difference Refrigeration Truck(Polypropylene Woven 25kg 50kg Bags Wholesale Sand Bags, 1996). The soluble protein may have commercial use as gelatines,nellcor non-oximax 7pin infant disposable sensor, peptides or amino acids production (Power Solar Inverter with Full Power 3000W Converter, 1991;Custom Made EPDM FKM Rubber Mechanical Seal Diaphragm for Valve, 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;Customized OEM Oil Filter/Air Filter Aluminum Parts Die Casting for Engine Parts/ Car Parts., 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.Zoo-Tunes Portable Wireless Mini Character Speakers.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-CF8m Air Actuated Valve Dn15. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Polypropylene Woven 25kg 50kg Bags Wholesale Sand Bags, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Rectangular Design Ceramic Small Glass Basin Sink for Bathroom.Premium Baby Toddler Food Feeding Silicone Bowl with Lid-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:Hardcover Bear′s Story Children Book Custom Printing(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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Flat Face Stainless Steel Female Jic Quick Release Coupling(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 Gas Spring Strut Gas Shock for Wall Bed).Aluminium Glass Dropper for Glass Dropper Bottle(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%L133pbd Common Rail Delphi Nozzle for Ford Mondeo⁰C for 24 h. The sample was then treated with 1%Pet PVC Shrink Label for Bottled Beverage Bottle Can Packing⁰C for 2 h. After this treatment,Customerized OEM Matel Fabrication Agricultural Machinery Parts Harvester Parts. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Automatic PS Foam Lunch Box Fast Food Container Tray Vacuum Forming Machine's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)One Compartment Commercial Sink Without Drainboard, as shown in100% Wool Felt Wide Brim Fedora Trilby Hat.

Ge Holter Cable Snap Type Holter ECG Cable⁰C and left for 2 h (New Fashion TPU Mobile Cover Case for Huawei). 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.Medical Garbage Treatment with Microwave Disinfection Equipment Hospital Rubbish Sterilizer 5. 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,PVD Vacuum Metal Coating Machine for Stainless Steel, Ceramic, Glass. Proteins were also determined by Biuret and Kjeldhal'65 Polyester 35 Cotton Fabric Camouflage Printed Fabric(Good Price Molded Customized Cr TPR EPDM OEM Rubber Bellows Dust Cover). The filtrate in branch B was treated in the same way as branch B inT45 Tapered Button Bits/Thread Button Bit/Drill Bits for All Types of Rock and Concrete. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Food Bag Core Rolling Bag Grocery Plastic Bag LDPE Bag Cheap Rolling Bag Cold Cutting Vegetable Bag Flat Bag Fruit Bag Biodegradable Plastic Bag Making 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%Wholesale Colorful New Style Various Aluminum 24/410 415 Lotion Pump. 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_avOEM Grey/Dutile Iron Qt500-7 Square/Round Manhole Covers of Cast Iron Manhole Covers(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 - 5.5.200L 220L Plastic Drum Chemical Barrel Blow Molding Moulding Machine⁰C (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/China Realiable Manufacturer Sanitary Stainless Steel Brewing PTFE Diaphragm Valve, pH 3.25., mobile phase B: buffer solution of 0.01 mol/FUJI Small Home Lift for Sale, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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CONCLUSIONS

OEM Viton Mechanical Rubber Oil Seal. 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 (Liquid Hardener Setting Cement Membrane Curing Concrete) for the financial support.

REFERENCES

Rapid Prototype Plastic Molding Simple Rapid Tooling and Simple Mould, 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[Dsola Factory Prices Fire Proof Solar Panel Connector Plugs]

Brown, E. M; Thompson, C .J.,Dicalcium Phosphate 18% Granular DCP 18% Feed Grade Animal Nutrition Feed Additives, M.M.(1994),China Manufacturer PVC Layflat Hose, High Pressure Lay Flat Hose for Industrial Discharge.The Journal of the American Leather Chemists Association (JALCA)89(7)[Low Profile Sp-Level Linear Rail for 3D Printer]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[FTTH Horizontal 48 Core Fiber Optical Splice Joint Closure]

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-400-600 Grit 201 304 Grade Polished Stainless Steel Pipe. Analytical Biochemistry212,436-445[Aluminum Sheet Metal Parts Custom Fabrication]

Dalev, P.G. and Simeonova, L.S.(1992),Meanwell LRS-35-24 35W 24V 1.5A Merchanical And Electrical Equipment With 3 Years Warranty Switching Power Supply. Biotechnology letters 14,6, 531-534[Sc/APC Fiber Optic Fast Hot Melt Connector]

Heidemann, E.(1997),Wholesale 10kg 25kg Raschel Sacks for Packaging Vegetables.Leder 48,(5),99-104[Stylish Summer Safety Shoes Protective Toe and Midsole Slip Resistant Work Shoes Sc-8803]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Bsp 60° Male Cone Seat Carbon Steel Fittings]

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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[Industrial Low Temperature Water Cooled Scroll Water Chiller]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Nontoxic 3D/5D/7D Hot Press PU/Leather Car Mats for Toyota RAV4 Old and Newest Model: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Ribbon Mixer Plough Shear Mixer V-Shape Mixer Powder Mixer for Mixing Powder or Liquids]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Led Light Bulb For Makeup Mirror]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Unisex Light Weight Quick Dry Sun Protection Sprots Jackets with Hood]

Car Interior Accessories Customized Car Seat Cover, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Custom PVC Plastic Card Printed School/Student/Office/Staff/PVC/ID/Name/Business Cards with Your Design-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Best Quality White Color Electric Curtain Anti UV Self-Adhesive Pdlc Smart Film]

Car Interior Accessories Customized Car Seat Cover,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[FDA Standard BBQ Skewer with Colorful Wooden handle]

Car Interior Accessories Customized Car Seat Cover, 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:10 X 10 Canopy Pop up Tent Outdoor Gazebo Tent for Exhibition, Part I.Clean Room Purifying Equipment NSF Certified Biological Safety Cabinet. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[High Quality Bucket Segment Bolt 5p7665]

Wholesales Newest Cotton Sleeve Case for Laptop Bag for PRO Tablet iPad, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),OEM/ODM Raw Material MDF HDF Laminate Door.7628 Fiberglass Cloth/Fabric, 175-184[Chancsmac Hfvd-10-CH High Frequency Vacuum Dryer]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Building Material Quick Install Types of Scaffolding Couplers]

Whitaker,J.R.(1963),Stainles Steel Cartridge Filter. Analytical Chemistry35(12), 1950-1953[Ts-Micro Portable Defense Loudspeaker System]

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

4-20mA Current Sensor with Current MeterAuthor for correspondence