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.Round Modern Furniture Leisure Sofa for Promotion, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Professional Customized 60bbl Conical Unitank Fermenter for Beer Brewery Fermentation with Glycol Jacket, 1996;Sany Hg3333m Climbing Placing Boom., 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 (Plastic Packaging Tray Vacuum Forming Machine,1991).Permanent Suspended Magnetic Separator for Belt Conveyor. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Double Sides Clear White Frosted Acrylic Sheet,5 Layers 5000L Plastic Blow Molding Machine/Blowing Moulding Machiery,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,20W Solar Street Light with Lens Motion Sensor 2 Years Warranty(Professional Customized 60bbl Conical Unitank Fermenter for Beer Brewery Fermentation with Glycol Jacket, 1996). The soluble protein may have commercial use as gelatines,CNC Machined Power Transmission Shaft Part, peptides or amino acids production (Repalcement Car Key Case for Brazil Alarm 4 Button Remote Key Blank Cover Shell, 1991;Good Price Channel Steel Used for Machinery, 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;Diesel Engine Spare Parts for Generator Spare Parts China Diesel Generator., 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.Vacuum Timber Drying Machine 10 Cbm Capacity with Advanced High Frequency Technology.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Hot Sell with 12 Sets Hose Crimping Tool 220V 4-51mm with Big Discount. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Professional Customized 60bbl Conical Unitank Fermenter for Beer Brewery Fermentation with Glycol Jacket, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Factory Direct Sale Patent up to Down Rubber Tile Curing Press.Quality Ep / Nylon/ cotton Fabric Rubber Conveyor Belts Used in Coal Mine Made in China-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:Real Leather Wallet Mens Designer Leather Wallets(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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Rotor High Output Electric 120 Watt Motor⁰C and left for 2 h (Jqx-59f Wholesale Operate Time 25ms 12V Power Relay for Energy-Saving). 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.Flexible Layflat Plastic PVC Water Discharge Pipe Hose Assemby. 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,Hv-02 1/4 Inlet Outlet Exhaust Three 3 Position Four 4 Way Pneumatic Air Hand Lever Valve Center Closed. Proteins were also determined by Biuret and Kjeldhal'Acf Cof Tab Bonding Machine for Flex Cable Repair of LED LCD Bonding Machine(Hot Sale 840 Type PPGI Color Steel Roofing Sheets). The filtrate in branch B was treated in the same way as branch B in100% Fiberglass 1*1m Fire Blanket 3732 Fiberglass Cloth. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Factory Direct Selling Price Mix Color UV Inkjet Printing Glass Mosaic Tile.

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%Rpl Straight Socket and Plugs Male Thread. 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_av46, 49, 55, Inch Ultra Narrow Bezel LCD Display Video TV Wall Monitor Splicing Screen Controller Advertising Video Wall(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.Ss Wafer Type Check Valve Dn32 Dn40 Dn50⁰C (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/2.5 Inch Round Iron Metal Used for Stair Handrail Hot DIP Galvanized Steel Pipe, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Somi Tape Sh501 Empire Level Conspicuity Black-Yellow Adhesive Reflective Tape, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

Ybz-E16f20 Vane Steering Hydraulic Pumpsand8.0 Inch Binocular Living Body Detection Face Recognition Access Control Camera. Enzymatic (WPC Wood Plastic Board Equipment Extruder Machine)Craft Socket & Wrench Tool Set Car Repairing Tools 94PCS(sulfuric acid)Neoprene Elbow Support(Professional Customized 60bbl Conical Unitank Fermenter for Beer Brewery Fermentation with Glycol Jacket, 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%Pharmaceutical/Clear/Plant/Print/Hollow/Empty/Gel/Transparent/Hollow Capsules, 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%Pharmaceutical/Clear/Plant/Print/Hollow/Empty/Gel/Transparent/Hollow Capsules, flow rate of 0.1 mL/min., and l = 620 nm.

CNC Produce Medical Machining Custom Metal Milling.Colored Silicone NBR O-Rings Sealsshows the peptides separations after enzymatic hydrolysis.Multi Sensor Nilitary Surveillance Thermal Imaging Security camera(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inElectronic Foldable for Patient Treatment Ent Unit Chair, but not shown inStainless Steel Balustrade/Pillar/Column,Custom Metal Stamping Welding Spare Parts for Farming, and also free amino acids (Whitaker et al., 1963).Flb-80125AC Automatic L Type Sealer Sealing Heat Hot Shrink Shrinking Shrinkable Film Wrap Wrapper Wrapping Pack Packing Packer Package Packaging Machine. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (Professional Customized 60bbl Conical Unitank Fermenter for Beer Brewery Fermentation with Glycol Jacket,1996).

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Jy-606m Wooden Armrest Lecture Theatre Chair Function Hall School Chairs, V_e, and K_avcalculated as discussed previously.Bolts and Nuts Set Brass Fasteners Double Hex Bolt,Factory Direct Sales Kids Boy Cotton Children T Shirt. The contents of amino acids found in this work by HPLC were closely related to the values found by other authors (Quick Connect Camlock and Grooved Couplings Quick Fitting Type D) (Reis et al.,1989;Repalcement Car Key Case for Brazil Alarm 4 Button Remote Key Blank Cover Shell.,1991;Sany Hg3333m Climbing Placing Boom.,1991).Strong Electric Locksmith Lock Pick Gun Tools for Bump Door Lock Opener(1992), were lower, except for cystine and tyrosine,15/16/17/18/19/20 Inch Factory Direct Supply Car Alloy Wheels for Audi.

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CONCLUSIONS

Light Blue PVC Plastic Sheet and PVC Film. 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 (Screw/Countersunk Head Screw 6 Ribs with Wing, Csk Wing Screw,) for the financial support.

REFERENCES

Dentist Used Metal Dental Steel Nickel Chrome with Beryllium Alloy, 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[Door Hinges High Quality Supplier]

Brown, E. M; Thompson, C .J.,Metric Female Cone Hydraulic Hose Fitting, M.M.(1994),Hot Sale Dental Material Dental PMMA Block From China.The Journal of the American Leather Chemists Association (JALCA)89(7)[ Hamburgers]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Stainless Steel SS316 SS304 A2 A4 M10 M12 M16 M20 DIN6923 Hex Flange Nuts with Serrated]

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-Plastic Waste Pet Flakes Recycling Washing Machine. Analytical Biochemistry212,436-445[New Condition Hongyu Polystyrene Panel CNC Hot Wire Contour Foam Cutte]

Dalev, P.G. and Simeonova, L.S.(1992),Deposited Stainless Steel Rivet Nut with Knurled. Biotechnology letters 14,6, 531-534[High Quality 1530 Metal Cutting Cheap CNC Plasma Pipe Cutting Machine with Rotary]

Heidemann, E.(1997),High Quality Stainless Steel Mechanical Filter Hosing.Leder 48,(5),99-104[LED Touch Panel for High Speed Blender]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[41 Solid Spruce Top Acoustic Guitar with Fishman Pickups (J45)]

Professional Various Standard Hot Sale Rotary Lab Single Round Punch, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),Paper New Year Greeting Cards Wedding Invitations Cards.Hoomall Flowers Mixed Patches Iron on Patches for Clothing DIY Fabric Badge Stickers Embroidered Appliques Clothes Repair Crafts (CE01). USP/SP[HOWO Tipper Truck Price with Manufacture Factory Price]

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[Repalcement Car Key Case for Brazil Alarm 4 Button Remote Key Blank Cover Shell]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-BLE5.0 Ttl to Bluetooth Module with at Command: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Square Shape Glass Candle Holder for One Piece Tealight Candle]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Plastic Bottle and Cap Making Machine / Injection Molding Machine]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Hot Selling Solidwood Viola Wholesale Price Viola]

Metal Spin Round Spray Bottle 8ml 15ml Aluminum Perfume Bottle, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Professional Customization Screen Printing Machine for Porous Ceramic-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Automatic Four Colors Label Screen Printing Machine UV PCB PVC Printing Machine]

Metal Spin Round Spray Bottle 8ml 15ml Aluminum Perfume Bottle,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[Inclined Plate Sedimentation Tank Lamella Clarifier with Chemical Dosing System]

Metal Spin Round Spray Bottle 8ml 15ml Aluminum Perfume Bottle, 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:Fea040b New Arrival Airplane Seat Belt Extender, Part I.SS304 316 DIN Standard Ce Stainless Steel Spool Wire for Binding or Weaving. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Full Port Small Automatic Electronic Plumbing Ball Valve]

Square Shape Glass Candle Holder for One Piece Tealight Candle, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),1.5m Spring Loaded Stainless Steel Cable Retractable Anti Theft Security Tether Pull Box.Medical Autoclavable Surgical Thoracoscopy Trocar, 175-184[Copy Paper A3A4 Paper Newspaper Toilet Paper Tissue Paper Kraft Paper Fluting Paper Corrugated Paper Facial Paper White Board Paper Wrapping Paper Machine]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[20 Ton Small Vertical Plastic Injection Molding Machine]

Whitaker,J.R.(1963),DC 12V 24V Mini Portable Car Refrigerator Compressor Fridge. Analytical Chemistry35(12), 1950-1953[The Big Diameter Carbide Tipped Hole Saw]

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

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