To determine the cellulolytic activities of the tested bacteria, cultures were grown overnight (OD600 ∼0.5) and spot-inoculated in triplicates of 5 µL on plates with solid Mandels and Reese medium (Mandels and Reese, 1957 (link)) containing carboxymethyl cellulose (CMC) sodium salt (Sigma-Aldrich) (Liang et al., 2014 (link)). Bacteria were then incubated at 20°C for 7 days. After the incubation time, plates were flooded with an aqueous solution of Congo red (1 mg/ml) for 15 min. The Congo red solution was then poured off and visualised zones of hydrolysis were stabilised by flooding the agar with 1 M HCl (Teather and Wood, 1982 (link)).
Cmc sodium salt
Manufactured by Merck Group
Sourced in United States, Germany
CMC sodium salt is a white, odorless powder that is used as a suspending agent, thickener, and stabilizer in various pharmaceutical and industrial applications. It is a cellulose derivative that is water-soluble and can be used to modify the viscosity and texture of aqueous solutions and dispersions.
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Lab products found in correlation
Dodecylbenzene sulfonic acid, by Merck Group (1 mentions)
Aniline, by Junsei (1 mentions)
Acetonitrile, by Junsei (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
Yeast extract, by BD (1 mentions)
Peptone, by BD (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Sodium sulfate, by Merck Group (1 mentions)
Potassium hydroxide, by Merck Group (1 mentions)
Potassium nitrate, by Merck Group (1 mentions)
Sodium borohydride, by Merck Group (1 mentions)
Methacrylic anhydride, by Merck Group (1 mentions)
Lithium phenyl 2 4 6 trimethylbenzoylphosphinate lap, by Merck Group (1 mentions)
6 protocols using cmc sodium salt
1
Screening for Cellulolytic Bacterial Strains
2
Polymer Characterization for Material Synthesis
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MC powder with a viscosity of 4,000 cP (in 2% solution) and HPMC powder with a viscosity of 4,260 cP (in 2% solution) were obtained from LOTTE Fine Chemical Co., Ltd (Incheon, Korea). CMC sodium salt with a viscosity of 50 to 200 cp (in 4% solution) was obtained from Sigma Chemical CO., (St. Louis, MO, USA). Yeast extract and peptone were purchased from BD Biosciences (San Jose, CA, USA). Glucose, hydrogen peroxide (34.5%), sodium hydroxide, ethyl alcohol (99.9%) and acetic acid were supplied by Duksan Pure Chemical Co., Ltd. (Seoul, Korea). Dodecylbenzene sulfonic acid (DBSA) and trifluoroacetic acid (TFA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Aniline and acetonitrile were purchased from Junsei Chemical Co., Ltd. (Tokyo, Japan). Ammonium peroxydisulfate (APS) was supplied by Kanto Chemical Co., Inc. (Tokyo, Japan). All chemical reagents were used as received.
3
Synthesis of Hydroxypropyl Cellulose from Sugar
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Commercially available sugar was used as the starting material for the HTC synthesis. PFOA (96%) and CMC sodium salt with a molecular weight of 90 kDa, a polymerization degree of 400, a substitution degree of 0.65–0.90 with a sodium content of approximately 8 wt.-% and a purity of 99.5% was purchased from Sigma-Aldrich (St. Louis, MO, USA). MCB (≥99%), sodium chloride, sodium sulfate, sodium borohydride, sodium hydroxide, hydrochloric acid (37%) potassium hydroxide and potassium nitrate were all analytical grade and purchased from Merck (Darmstadt, Germany). Potassium hydrogen phthalate (p.a.) was purchased from Riedel-de Haën (Seelze, Germany).
4
Eucalyptus Pulp-Based Biomaterials
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Bleached kraft Eucalyptus globulus pulp, air dried, was obtained from an industrial pulp mill and hydrated at FibEnTech Research Unit (University of Beira Interior, Covilhã, Portugal) using distilled water. CMC sodium salt, high viscosity (1500–3000) cP in 1% H2O (25 °C) was purchased from Sigma Aldrich (Darmstadt, Germany). Alginate sodium salt, with high viscosity, was acquired from BDH Chemicals Ltd. (London, UK). Tea tree essential oil was purchased at a local store. The microalgae Chlorella vulgaris was obtained from Aqualgae (Viana do Castelo, Portugal). All other chemicals and reagents used in the study were analytical grade.
5
Cytotoxicity Evaluation of CMC Hydrogel
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Medium viscosity CMC sodium salt, methacrylic anhydride, and lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) were purchased from Sigma Aldrich. For a cytotoxicity test, DMEM purchased from Carlo Erba (Italy) was used.
6
Characterizing Eucalyptus and Pine Pulps for MFC Production
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An industrial never-dried, total chlorine-free (TCF) bleached Eucalyptus kraft pulp (hardwood) was used in this study, with a Shöpper-Riegler degree (°SR) of 25°. Additionally, an industrial air-dried, elemental chlorine-free (ECF) bleached Pinus kraft pulp (softwood) was subject to a mechanical treatment of refining using a lab-scale PFI-mill beater to reach the same target of 25 °SR, according to ISO 5264/2. CMC sodium salt with high viscosity (1500–3000 cps) in 1% H2O (25 °C), an average molecular weight of 700,000 g/mol, and a degree of substitution of 0.65–0.90 (carboxymethyl group per anhydroglucose unit) was purchased from Sigma Aldrich (Darmstadt, Germany) and used in this study as an additive. Eucalyptus essential oil (EO) was purchased from Medical Plants Department store, physalis®. To produce the MFC sample, an industrial air-dried, ECF bleached Eucalyptus kraft pulp was used.
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