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4 protocols using phenanthrene

1

Phenanthrene Degradation by Engineered Pseudomonas

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Glass and polystyrene were cut into 1 cm ×1 cm pieces, sterilized with 70% ethanol for 15 min, and then rinsed with sterile water before use. Stock strain JM2-gfp was from phenanthrene-degrading bacterium Pseudomonas sp. JM2, labeled with GFP as visual marker [23 (link)].
Analytical grade phenanthrene (≥97%) was purchased from Tokyo Chemical Industry Co., Ltd. (Shanghai, China). Methyl alcohol, n-hexane and acetone of HPLC grade were purchased from J&K Scientific Ltd. (Beijing, China).
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2

Biosurfactant Rhamnolipid for Hydrophobic Compound Removal

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The biosurfactant rhamnolipids was purchased from Hunan Zhijin Biotech Co., Ltd., Changsha, China. The biosurfactant was originally received as a blend of 2/3 mono-rhamnolipid and 1/3 di-rhamnolipid, and no further alteration was performed. It is a brown viscous liquid with approximate purify of 40%. The nonionic surfactant TX100 was with purity higher than 98%, and phenanthrene was with purity greater than 97%, both substances were obtained from Tokyo Chemical Industry Co., Ltd., Tokyo, Japan. Acetic acid was purchased from J&K Scientific Ltd., Beijing, China. The selected properties of above substances are shown in Table 1.
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3

Procurement of Diverse Chemical Compounds

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Abietic acid, ADD, DHEA, methyltestosterone,
phenanthrene, and progesterone were purchased from Tokyo Chemical
Industry (Tokyo, Japan). DehydroAbietic acid, dibenzothiophene, 7-ethoxycoumarin,
ibuprofen, and testosterone were purchased from Fujifilm Wako Pure
Chemical (Osaka, Japan). Pyrene was purchased from Sigma-Aldrich Japan
(Tokyo, Japan). Diclofenac was purchased from Combi-Blocks USA (San
Diego, CA, USA). Ferruginol was kindly provided by Dr. H. Suhara (Miyazaki
Prefectural Wood Utilization Research Center, Japan). Yeast nitrogen
base without amino acids was purchased from Formedium (Hunstanton,
UK). Dropout supplements (DOS) were purchased from TaKaRa Bio USA
(Mountain View, CA, USA). Custom-synthesized oligonucleotide primers
were obtained from Sigma-Aldrich Japan. All other chemicals were of
reagent grade. Deionized water was obtained using a Barnstead Smart2Pure
System (Thermo Fisher Scientific, Waltham, MA, USA).
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4

HPLC Analysis of Diverse Organic Compounds

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APS (particle size, 5 µm; pore size, 120 Å) was a kind gift from Daiso Chemical (Osaka, Japan). HPLC grade of acetonitrile (ACN), benzene, naphthalene, anthracene, toluene, uracil, thymidine, cytosine, uridine, adenosine, thymine, cytidine, theobromine and caffeine were obtained from Wako Pure Chemicals (Osaka, Japan). PLLA (average molecular weight: 5,000) and dioxane was from Nacalai Tesque (Kyoto, Japan).
Hypoxanthine was purchased from Merck KGaA (Darmstadt, Germany).
4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM), inosine, phenanthrene and pyridoxine were from Tokyo Chemical Industry (Tokyo, Japan). Ethylbenzene and adenine were from Kishida Chemicals (Osaka, Japan).
Paraxanthine was obtained from Sigma (St. Louis, MO, USA).
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