M. spathulata R25L270 strain was obtained from the culture collection of the Universidade Federal de Minas Gerais (Belo Horizonte, Brazil). Macaúba cake was obtained from Biodiesel Company (Minas Gerais, Brazil). The fish oil was obtained from BTSA, Biotecnologías Aplicadas, S.L. (Madrid, Spain), DEAE-agarose (Diethylaminoethyl-agarose), Octyl-Sepharose™ CL-4B, Phenyl-Sepharose™ CL-4B, Butyl-Sepharose, Sulfopropyl-Sepharose and Cyanogenogen Bromide (BrRCN)(CNBr) supports were obtained from GE Bio-Sciences AB (Uppsala, Sweden). The substrate pNPP and all other chemicals were obtained from Sigma Chemical Co. (St. Louis, MO, USA).
Octyl sepharose cl 4b
Manufactured by GE Healthcare
Sourced in Sweden
Octyl-Sepharose CL-4B is a type of chromatography media used for the purification and separation of biomolecules. It is a hydrophobic interaction chromatography (HIC) resin composed of octyl (C8) groups coupled to a crosslinked agarose matrix. This resin is designed to facilitate the separation and purification of proteins, enzymes, and other biomolecules based on their hydrophobic interactions with the octyl ligands.
Automatically generated - may contain errors
Lab products found in correlation
Phenyl sepharose cl 4b, by GE Healthcare (1 mentions)
Butyl sepharose, by GE Healthcare (1 mentions)
Sulfopropyl sepharose, by GE Healthcare (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Triethylamine, by Merck Group (1 mentions)
Sodium borohydride, by Merck Group (1 mentions)
Ethylenediamine, by Thermo Fisher Scientific (1 mentions)
Sodium periodate, by Merck Group (1 mentions)
Epichlorohydrin, by Merck Group (1 mentions)
Glutaraldehyde solution, by Thermo Fisher Scientific (1 mentions)
Iminodiacetic acid, by Merck Group (1 mentions)
Bradford reagent, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
2 propanol, by Merck Group (1 mentions)
Trisodium citrate, by Chempur (1 mentions)
Tris base, by Merck Group (1 mentions)
N heptane, by Merck Group (1 mentions)
R flurbiprofen, by Merck Group (1 mentions)
5 protocols using octyl sepharose cl 4b
1
Macaúba Cake and Fish Oil Enzyme Purification
2
Immobilization and Characterization of Enzymes
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Agarose 4 BCL was purchased from Agarose Bead Technologies (Madrid, Spain). Epichlorohydrin, iminodiacetic acid, triethylamine, sodium borohydride, sodium periodate, 1-octanothiol, bovine serum albumin, O-nitro-phenyl-β-d -galactopyranoside (O-NPG), p-nitrophenyl proprionate (p-NPP), 4-nitrophenyl-β-d -glucopyranoside (p-NPG), and high molecular weight protein (Sigma MarkerTM, St. Louis, MO, USA) were purchased from Sigma (Sigma-Aldrich®, St. Louis, MO, USA). Glutaraldehyde solution (25%, v/v) and ethylenediamine were purchased from Alfa Aesar (Thermo Fisher Scientific®, Waltham, MA, USA). Octyl-Sepharose CL-4B was purchased from GE Healthcare Bio-Sciences (Uppsala, Sweden). The β-galactosidase from Kluyveromyces lactis (Lactozym pure 6500 L) (KlBgal) was kindly supplied by Novozymes A/S (Copenhagen, Denmark). The lipase from Candida rugosa (CRL) was purchased from Sigma (Sigma-Aldrich®, St. Louis, MO, USA). Overexpression and purification of lipase (LipC12) and β-glucosidase (EaBglA) were performed as previously described by Glogauer et al. [40 (link)] and Crespim et al. [47 (link)], respectively. All other chemicals used were of analytical grade.
3
Enzymatic Enantioselective Flurbiprofen Synthesis
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Octyl-Sepharose CL-4B (GE Healthcare, Uppsala, Sweden), (R,S)-flurbiprofen, (R)-flurbiprofen, methanol, n-heptane, 2-propanol, trifluoroacetic acid, 1,2-dichloropropane (DCP), diisopropyl ether (DIPE), tert-butyl methyl ether (MTBE), Bradford reagent, hydrochloric acid, and Tris base were purchased from Sigma-Aldrich (Steinheim, Germany). Dichloromethane (DCM), 1,2-dichloroethane (DCE), citric acid, disodium hydrogen phosphate dihydrate, monosodium hydrogen phosphate monohydrate, and molecular sieves 4 Å were gained from POCH (Gliwice, Poland). Trisodium citrate was from Chempur (Piekary Ślaskie, Poland). The lipase B from Candida antarctica (CALB, produced in yeast) was from ChiralVision (Leiden, The Netherlands), and the lipase from Candida rugosa (CRL-OF) from Meito Sangyo (Tokyo, Japan).
4
Arabinose Metabolism Analysis via GC/MS
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Most of the reagents were bought from Sigma-Aldrich and used without further purification. The silylation agent, HTP-trisil, was bought from Thermo-Fisher Scientific. Solvents were purchased from Fisher Scientific. Uniformly labeled 13C5-D-arabinose was purchased from Cambridge Isotope Laboratories, Inc. Octyl sepharose CL 4B was obtained from GE Healthcare. GC/MS was recorded on CP 3800 gas chromatograph (Varian) equipped with an MS320 mass spectrometer. Helium was used as the carrier gas with a flow rate of 1 ml/min. Argon was used as the collision gas in MS/MS experiments and methane was used as the chemical ionization gas for the negative chemical ionization analyses. The samples were analyzed on an Agilent J&W capillary column VF-5ms column (30 m × 0.25 mm i.d.). The data analyses were carried out on a Varian WS data station. NMR spectroscopy was carried out on Innova 400 (Varian) at the Central Instrument Facility, CSU. Healthy nonendemic urine (NEU) samples were collected from willing co-worker-volunteers in CSU. The primary antibody CS-35, used in the dot blot assay was developed in our laboratory and the detection antibody anti-mouse IgG alkaline phosphatase was purchased from Sigma-Aldrich
5
Enzymatic Activity Characterization
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A TLL liquid formulation with 20.77 mg protein/mL was utilized in this paper, while lipase B from Candida antarctica (CALB) was a liquid formulation with 7.7 mg protein (kindly donated by Novozymes Spain (Madrid, Spain). Bradford’s reagent (utilized to calculate the protein concentration [98 (link)]), p-nitrophenyl-butyrate (p-NPB), triacetin, R- and S-methyl mandelate, acetonitrile for HPLC (gradient grade, ≥99.9%), glutaraldehyde (GA) solution (25% in H2O), ethylenediamine (EDA), picrylsulfonic acid (TNBS), polyethylenimine (PEI, MW 25,000), dextran sulfate (DS, MW 20,000), N-3-(dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (ECD), NiCl2, MgCl2, CoCl2, CuCl2 and ZnCl2 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Octyl Sepharose® CL-4B was acquired from GE Healthcare (Uppsala, Sweden). All other reagents were of analytical grade.
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