Pseudomonas aeruginosa strains used in this study are listed in Table 1 . Bacteria were grown in an Air-Jacketed Incubator IC802 (Yamato Scientific Co., Ltd., Tokyo, Japan) at 37 °C under aerobic conditions, as previously described [12 (link)]. Unless otherwise indicated, bacteria were cultured using lysogeny broth, Lennox (LB)-agar prepared fresh from 1.0% BactoTM Tryptone (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 0.5% BactoTM yeast extract (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), and 0.5% NaCl (FUJIFILM Wako Pure Chemical Corp., Osaka, Japan). Pseudomonas agar F was prepared from 2.0% BactoTM proteose peptone no. 3 (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 1.0% BactoTM Casitone (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 1.0% glycerol (Nacalai Tesque Inc., Kyoto, Japan), 0.15% K2HPO4 (FUJIFILM Wako Pure Chemical Corp., Osaka, Japan), 0.073% MgSO4⋅7H2O (FUJIFILM Wako Pure Chemical Corp.), and 1.5% agar (FUJIFILM Wako Pure Chemical Corp.), and used as a solid medium in STAR SDish9015 ver.2 petri dishes (Rikaken Co., Ltd., Nagoya, Japan) for assays of pyoverdine production. Bacterial growth was quantified by measuring the optical density at 600 nm (OD600) using a WPA CO8000 Cell Density Meter (Biochrom Ltd., Cambridge, UK).
K2hpo4
Manufactured by Fujifilm
Sourced in Japan
K2HPO4, also known as dipotassium hydrogen phosphate, is a chemical compound commonly used in laboratory settings. It serves as a buffering agent, maintaining a specific pH level in aqueous solutions. The compound is a white, crystalline powder that is soluble in water.
Automatically generated - may contain errors
Lab products found in correlation
Kh2po4, by Fujifilm (8 mentions)
Mgso4 7h2o, by Fujifilm (5 mentions)
Cacl2, by Fujifilm (5 mentions)
Nh4cl, by Fujifilm (4 mentions)
Fecl3 6h2o, by Fujifilm (4 mentions)
Sodium chloride (nacl), by Fujifilm (3 mentions)
Formic acid, by Fujifilm (3 mentions)
Na2hpo4 12h2o, by Fujifilm (3 mentions)
Potassium chloride (kcl), by Fujifilm (2 mentions)
N decane, by Merck Group (2 mentions)
Milli q system, by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Fujifilm (2 mentions)
Silica gel, by Merck Group (2 mentions)
Tetrahydrofuran, by Fujifilm (2 mentions)
Tween 80, by Fujifilm (2 mentions)
Sodium ascorbate, by Fujifilm (2 mentions)
Aniline, by Fujifilm (2 mentions)
Tris 2 ethylhexyl trimellitate, by Tokyo Chemical Industry (2 mentions)
Distilled water hplc grade, by Fujifilm (2 mentions)
Phosphate solution, by Fujifilm (2 mentions)
14 protocols using k2hpo4
1
Cultivation and Characterization of Pseudomonas aeruginosa
2
Fabrication and Characterization of Solid-State Nanopores
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The following reagents were used in this study: poly(methyl methacrylate) (PMMA) substrate (Mitsubishi Rayon; Japan); KCl, K2HPO4, KH2PO4, and ethylenediamine tetraacetic acid (EDTA; Wako; Japan); 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and phosphocholine from egg yolk (EggPC) (Avanti Polar Lipids; Alabama); n-decane (Sigma-Aldrich; St. Louis). Buffered electrolyte solutions were prepared from ultrapure water. The ultrapure water was prepared with >18 MΩ cm water from a Milli-Q system (Millipore). Wild-type αHL (Sigma-Aldrich; St. Louis) was obtained as a monomer polypeptide isolated from Staphylococcus aureus in the form of a lyophilized powder and dissolved at a concentration of 1.0 mg protein/mL in ultrapure water. During use, samples were diluted to the desired concentration using a buffered electrolyte solution, and stored at 4°C. For measurements in the field, all samples were stored at room temperature.
3
Diverse Chemical Standards Characterization
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As test chemicals, anthraquinone (AQ), 2-ethylanthraquinone (2-EA), dibenzofuran (DF), and bis(2-ethylhexyl)phthalate (Bis) were purchased from Tokyo Chemical Industry (Tokyo, Japan). i-Erythritol, 2,2-dimethyl-1,3-propanediol, and putrescine as test chemicals, and acetonitrile and distilled water (HPLC grade, respectively), tetrahydrofuran (stabilizer free, special grade), formic acid, K2HPO4, KH2PO4, Na2HPO4 · 12H2O, NH4Cl, MgSO4·7H2O, CaCl2, FeCl3 · 6H2O, 0.5% phosphate solution, α-D-Glucose, and 1 M sodium hydroxide were purchased from FUJIFILM Wako Pure Chemical (Osaka, Japan). Aniline as a test chemical and acetone were purchased from Kanto Chemical (Tokyo, Japan). Peptone was purchased from Showa Chemical (Tokyo, Japan). Silica gel (5 to 25 µm particle size for thin-layer chromatography) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium sulfite was purchased from Nacalai Tesque (Kyoto, Japan).
4
Recombinant Protein Production in E. coli
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BP1 was produced by a fermentation process using E.coli, which was previously reported in the literature28 ,44 and provided in an unprocessed powder form by Spiber Inc. The amino acid composition of BP1 was alanine (16.4%), tyrosine (12.0%), glutamine (23.7%), glycine (21.9%), proline (15.3%), serine (9.3%) and others (1.4%). The protein sequence structure is shown in Fig. 1 .
KH2PO4, K2HPO4, NaCl, Na2HPO4·H2O, NH4Cl, MgCl2·6H2O, CaCl2, FeCl3·6H2O, Yeast extract, and agar powder were purchased from FUJIFILM Wako Pure Chemical Co. (Osaka, Japan). Na2SO4, KCl, HCl, Na2SO4, NaHCO3, and NaOH were purchased from Kanto Chemical Co., Inc. (Tokyo, Japan). Plysurf was purchased from DKS Co. Ltd. (Kyoto, Japan). All chemicals were of reagent grade and used without further purification. Pronase E (P5147 Protease Type XIV from Streptomyces griseus) was purchased from Sigma Aldrich, Proteinase K was purchased from TaKaRa Bio Inc (Kusatsu, Japan), and chymotrypsin was purchased from NACALAI TESQUE, INC (Kyoto, Japan).
KH2PO4, K2HPO4, NaCl, Na2HPO4·H2O, NH4Cl, MgCl2·6H2O, CaCl2, FeCl3·6H2O, Yeast extract, and agar powder were purchased from FUJIFILM Wako Pure Chemical Co. (Osaka, Japan). Na2SO4, KCl, HCl, Na2SO4, NaHCO3, and NaOH were purchased from Kanto Chemical Co., Inc. (Tokyo, Japan). Plysurf was purchased from DKS Co. Ltd. (Kyoto, Japan). All chemicals were of reagent grade and used without further purification. Pronase E (P5147 Protease Type XIV from Streptomyces griseus) was purchased from Sigma Aldrich, Proteinase K was purchased from TaKaRa Bio Inc (Kusatsu, Japan), and chymotrypsin was purchased from NACALAI TESQUE, INC (Kyoto, Japan).
5
Microbial Inactivation Assay Protocol
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The sanitizers used in this study were chlorous acid water (Honbusankei Co., Ltd.) and NaClO (Nankai Chemical Co., Ltd.). Sodium thiosulfate, sodium chlorite, KH2PO4, K2HPO4, NaOH, N,N-diethyl-p-phenylenediamine (DPD) and sodium sulfate were purchased from Wako Pure Chemical Industries, Ltd. 3,3',5,5'-tetramethylbenzidine (TMB) was purchased from Tokyo Chemical Industry Co. Ltd. BSA (35% in PBS), polypeptone and meat extract were purchased from Sigma, Sumitomo Dainippon Pharma Co., Ltd. and Nacalai Tesque Inc., respectively. BSA, polypeptone and meat extract were added to microbicidal assays as an organic matter load.
6
Biomimetic Hydroxyapatite Microrods Synthesis
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Cyclohexane, CaCl2, K2HPO4, and KOH were purchased from Wako Pure Chemical Industries and were of analytical grade. The three types of surfactants, namely, Span 80 (sorbitan monooleate), Tween 20 (polyoxyethylene (20) sorbitan monolaurate), and Tween 80 (polyoxyethylene (20) sorbitan monooleate) (a hydrophilic surfactant, HLB value: 15) [25] , were reagent grade products of Wako Pure Chemical Industries. Commercially available HA (Apatite HAP, monoclinic, Wako Pure Chemical Industries) was used as the HA microrods, for comparison. KBr for infrared (IR) analysis was purchased from Wako Pure Chemical Industries. Four proteins, namely, bovine serum albumin (A3059), bovine γ-globulin (G5009), equine skeletal muscle myoglobin (M0630), and chicken egg white lysozyme (L6876), were obtained from Sigma-Aldrich. Poly(l -lactic acid) (PLLA) was a gift from Toyota Motor Corp. The PLLA properties included the weight average molecular weight: 1.22×105 (Mw/Mn = 3.0), optical purity: 98.5%, melting point: 174.0 °C, and glass transition temperature: 59.7 °C. Analytical grade 1,4-dioxane was purchased from Wako Pure Chemical Industries. All chemicals were used without further purification.
7
Anaerobic Bacteria Cultivation Protocol
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Reagents included De Man Rogosa and Sharpe (MRS) media (Becton Dickinson Difco, U.S.A.), hipolypeptone (Nihon Seiyaku, Japan), beef extract (MP Biomedicals, LLC, France), yeast extract (Becton Dickinson Difco), TBO (Waldeck, Munster), and Percoll (GE Healthcare Life Sciences, Japan). Glucose, Tween 80, K2HPO4, sodium ascorbate, L-cysteine-HCl, NaNO3, MgSO4, KH2PO4, NaH2PO4, NaCl, and 4′,6-diamidino-2-phenylindole (DAPI) were procured from Fujifilm Wako Pure Chemical Corporation, Japan. Data were collected using a spectrophotometer (UV-1200, Shimadzu, Japan) and a fluorescence microscope DMRXA/RD (Leica Microsystems, Germany). Cultures were prepared using fixed-type (model of rotor: BN 4–6) (H-201FR, Kokusan, Japan) and swing-type (model of rotor: RF 110) centrifuges (H-500FR, Kokusan). Deionised and doubly distilled water was used in all experiments.
8
Twitching Motility Observation on Minimal Medium
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Colonies on minimal medium (MM) [0.175% (w/v) K2HPO4 (Wako Pure Chemical Industries, Ltd.), 0.075% (w/v) KH2PO4 (Wako Pure Chemical Industries, Ltd.), 0.015% (w/v) sodium citrate (Wako Pure Chemical Industries, Ltd.), 0.025% (w/v) MgSO4.7H2O (Wako Pure Chemical Industries, Ltd.), 0.125% (w/v) (NH4)2SO4 (Wako Pure Chemical Industries, Ltd.), 0.5% (w/v) glucose, and 1.5% (w/v) agar (KATAYAMA Chemical Industries, Co. Ltd., Osaka, Japan)] (Addy et al., 2012 (link)) plates were examined for twitching motility by placing a Petri dish without its lid on the stage of an inverted microscope (an Olympus CKX41, Tokyo, Japan) equipped with 4× and 10× objectives.
9
Fabrication of Nanopore Membrane Devices
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The following chemicals were used without further purification: egg yolk phosphatidylcholine (EggPC; Avanti Polar Lipid, Alabaster, AL, USA); n-decane (Sigma-Aldrich, St. Louis, MO, USA); ethylenediaminetetraacetic acid (EDTA), KCl, K2HPO4, and KH2PO4 (Wako Pure Chemical Industries, Ltd., Osaka, Japan); wild-type αHL from Staphylococcus aureus (Sigma-Aldrich); and HPLC-grade DNA oligonucleotide (BEX Co., Ltd., Tokyo, Japan). Ultrapure water with >18 MΩ cm was prepared using a MilliQ system (Merck Millipore, Billerica, MA, USA). Poly(methyl methacrylate) (PMMA) plates with thickness of 0.075, 3.0, and 4.0 mm were obtained from Mitsubishi Rayon (Tokyo, Japan).
10
Biodegradability of Organic Chemicals
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Anthraquinone (AQ), 2-hydroxy-4-methoxybenzophenone (OxB), bis(2-ethylhexyl)phthalate (Bis), 2-ethylanthraquinone (2-EA), 2-hydroxy-4-n-octyloxybenzophenone (OB), and tris(2-ethylhexyl)trimellitate (Tris), shown in Table 1 , were purchased as test chemicals for ready biodegradability from Tokyo Chemical Industry (TCI, Japan). Distilled water (HPLC grade), formic acid, acetonitrile (HPLC grade), tetrahydrofuran (stabilizer free, special grade), K2HPO4, KH2PO4, Na2HPO4·12H2O, NH4Cl, MgSO4·7H2O, CaCl2, FeCl3·6H2O, 0.5% phosphate solution, and aniline were purchased from FUJIFILM Wako Pure Chemical (Japan); 1 M sodium hydroxide solution was purchased from Kanto Chemical (Japan); and chloroform was purchased from Nacalai Tesque (Japan). Silica gel (5 to 25 µm particle size for thin-layer chromatography) was purchased from Sigma-Aldrich (USA), and Tween 80 was purchased from TCI. Water solubility for some of the test chemicals was estimated using EPI Suite™ ver. 4.11, a Windows-based suite of physical/chemical property and environmental fate estimation programs developed by the US EPA and Syracuse Research Corp.14 )
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