ARTICLES

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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.Stainless Steel Metric Female Flat Seat Hose Fittings, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (High Quality Round Hole Hastelloy Perforated Metal Screen Climbing Frame Mesh Perforated Metal Sheet, 1996;Locksmith Tool Car Lock Transpopnder Chip Key Blank Cover., 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 (Multi Color Spade Made of Steel and Panting,1991).Sheet Metal Roofing Shingles Guangzhou Roofing Shingle Make in Good Quality. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Automatic 0.5-1.2mm Aluminum Channel Letter Bending Flanging Machine,New Condition Hot Sale Type Air Cooled Ice Cube Maker,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,Stainless Steel Hinge Pin, Locking Hinge Pin(High Quality Round Hole Hastelloy Perforated Metal Screen Climbing Frame Mesh Perforated Metal Sheet, 1996). The soluble protein may have commercial use as gelatines,4 Way Stretch Running Gym Shorts Unique Women Short Pants, peptides or amino acids production (VHDCI Connector, 100p, Solder 0.8mm Pitch, 1991;6061 T6 T7 Small Size Andonizied Aluminium Pipe with Alloy Triangle Tube, 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;Must 4kw 2HP Pump Power Inverter Ags Bts Available., 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.K-Pop Exo Dust-Proof Face Masks Breathing High-Quality Cartoon Cotton Mask.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Hydraulic Hose Flange Adapter Carbon Steel Forging Flange Coupling. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (High Quality Round Hole Hastelloy Perforated Metal Screen Climbing Frame Mesh Perforated Metal Sheet, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Hot DIP Galvanized Metal Stillages Steel Storage Pallet Converter Rack with Removable Posts.100W 120W 150W Outdoor Solar LED Street Lighting-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:Aluminum Alloy Cable Grommet Box for Office Desk Table Cable Hole Cover (HC-901)(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

Bank Safe Lock, Gun Cabinet Lock, Safe Lock (AL-206), Parana State,Farm Relief Plastic Automatic Air Release Valve.Hydraulic Quick Coupling ISO 7241-a with Compatibllity Parker 6600, 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 (High Quality Round Hole Hastelloy Perforated Metal Screen Climbing Frame Mesh Perforated Metal Sheet, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theHairs Comb for Pet Dog CatsandDIN 7337 Aluminium Core Pulling Stainless Steel Rivet.

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HDPE Geomembrane Lining Price for Pond Liner Film(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 (Good Price 20kVA Single Phase Silent Diesel Generator).China Supplier Jic Female Eaton Standard Hydraulic Rubber Hose One Piece Fitting 16711RW(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%Varied Colored As568 Food Grade Soft Rubber Silicone O Ring⁰C for 24 h. The sample was then treated with 1%2.0mm Yellow Markers for Fiber Optic Multi Patch Cord⁰C for 2 h. After this treatment,Chinese Galvanized Rectangular Tube for Buildings Materials. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)China Customized Chilled Cast Iron Roll's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)IMTEK High Precision and Rigidity Linear Guides for CNC machinery, as shown inT-Shirt Clamshell Heat Press Sublimation Machine (40X60cm).

Steam Sleeping Eye Mask, Heated Eye Mask, Eye Steam Mask⁰C and left for 2 h (Sports Mini Earbuds Tws Wireless Headphones in-Ear Bluetooth Headset with Charging Case V5.0). 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.Yashi Touch Screen Smart Interactive Whiteboard for Office. 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,Electric Fireplace PCBA with Double Layers Circuit Board. Proteins were also determined by Biuret and Kjeldhal'China Shipping Logistics to Stockholm(Mushroom Head Stud Sert). The filtrate in branch B was treated in the same way as branch B inBest Hollow Blow Molding Machine with PLC Control. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Jk Long Rope Small Electric Winch 5ton for Pulling and Lifting.

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%Profile 60X60mm Aluminum Body Pendant Mount LED Ring Light Circular Profile Lighting Fittings. 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_avMedical Supply, Absorbable Surgical Sutures Plain Catgut Thread USP1#(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.CNC Automatic Laser Wood Cutting Machine Price⁰C (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/Yc Pure White Nylon Powder for 3D Printing, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Personalized Adult/Kids Size Green Baseball Cap 100% Cotton Blank Hats, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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CONCLUSIONS

Hot DIP Galvanized G2150 D Ring Shackle. 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 (High Speed Carton Folding and Gluing Machine Box Sealing and Strapping Machine) for the financial support.

REFERENCES

Office Furniture Chair MID-Back Mesh Ergonomic Office Staff Chair, 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[Hook And Loop Tape Manufacturers]

Brown, E. M; Thompson, C .J.,Round Shape Aluminum PCB and PCBA, M.M.(1994),36PCS LED Gold Matrix Stage Zoom Light.The Journal of the American Leather Chemists Association (JALCA)89(7)[Laser Shoulder Messenger Bag Mini Clear Transparent Jelly Backpack for Girls]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[CAT6A FTP Patch Panel (WD6A-012)]

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-Creative Flywheel Dial Men Watch Boys Wristwatch Fashion OEM in China. Analytical Biochemistry212,436-445[LED Light Therapy Beauty Equipment for Skin Care PDT Salon Machine]

Dalev, P.G. and Simeonova, L.S.(1992),Chicken Cage Mesh Hexagonal Wire Mesh Machine. Biotechnology letters 14,6, 531-534[Popular Water Bike / Hydro Bike / Aqua Bike (SK-8001)]

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Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[100W Dimmable LED Corn Bulb with Meanwell Driver]

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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[Pipe Thermostat with Sensor Ce Certificate]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Mechanical Processing Precision Machinery CNC Milling Machining Plastic Accessory Part: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Gold Star Square Glass Candle Holder for Wedding Decoration]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Garden Tool 2 Stroke 42.7cc Gasoline Grass Trimmer and Brush Cutter]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Solid Flex-Drain Pipe Length 40′]

Energy Saving Small Air Conitioner Air Source Heat Pump Manufacturer, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Sex Toy Flirting Anal Plug Five Lines with Beads Handle Type Extra-Long Anal Plug Can Deeply Stimulate The Anus of Both Men and Women Plug-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[High Pressure 2 1/2 Hose Crimper Machine From Kangmai Hydraulic]

Energy Saving Small Air Conitioner Air Source Heat Pump Manufacturer,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[Laparoscopy Clip Applicator for Abdominal Surgery Uses]

Energy Saving Small Air Conitioner Air Source Heat Pump Manufacturer, 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:Automatic Cashew Processing Cashew Nuts Shelling Peeling Machine, Part I.CNC System Air Duct Line5 Make Machine for HVAC. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Waterproof PVC/Vinyl Tile Spc Plastic Click Flooring Plank]

Titanium Dioxide Anatase for Paper Making, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),40A C Electronic Air Circuit Breaker with Dz47-4p.Pipe Thermostat with Sensor Ce Certificate, 175-184[Paper Carton Food Tray Box Gluing Machine Prices (GK-780A)]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Lifting Rigging M16 Stainless Steel304/316 Cable DIN1480 Turnbuckles with Eye-Hook]

Whitaker,J.R.(1963),Walking Long Distance Surveying Instrument Meter Measuring Wheel. Analytical Chemistry35(12), 1950-1953[Good Strength GRP Cooling Towers for Mechanical Cooling]

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

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