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5 protocols using para cresol

1

Enzyme Purification and Characterization

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BPA, para-cresol (p-cresol), and other chemicals were obtained
from Sigma-Aldrich, and their purity was higher than 99%. Mushroom
tyrosinase (50 kU) was from Sigma-Aldrich as a solid with a specific
activity of 2430 unit mg–1 (1 unit is defined as
the enzyme activity resulting in an increase in absorbance at 280
nm of 0.001 at pH 6.5 at 25 °C in a 3 mL reaction solution containing l-tyrosine). Polyethylenimine (PEI, Mn 1200), poly(ethylene glycol) (PEG, Mn 1500), polyacrylate (PAL, Mn 1250),
diethylaminoethyl-cellulose (DEAE-cellulose, microgranular), carboxymethyl-cellulose
(CM-cellulose, microgranular), cellulose (fiber), hen egg-white lysozyme
(Mw: 14.4 kDa), and bovine serum albumin
(BSA, Mw: 66 kDa) were also from Sigma-Aldrich.
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2

Organosolv Lignin Tyrosinase Immobilization

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Organosolv lignin (OL) was purchased from Chemical Point (Oberhaching, Germany). Mushroom tyrosinase (1000 U/mg), bovine serum albumin (BSA), glutaraldehyde (GA), poly (diallyldimethylammonium chloride) (PDDA, 20% v/v water solution), 2-chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphopholane, chrome (III) acetylacetonate, deuterated chloroform (CDCl3), ascorbic acid, Bradford reagent, l-tyrosine, para-cresol, tyrosol, 4-hydroxyphenylacetic acid, 3-(4-hydroxyphenyl)propionic acid, catechol, potassium chloride (KCl), and organic solvents were purchased from Sigma–Aldrich (St. Louis, MO, USA), and were used without any further purification. All experiments were done in triplicate using native and immobilized tyrosinase in Phosphate buffer saline (PBS).
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3

Analytical Standards for Bioassays

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Ethanol, acetic acid, n-hexane, n-butanol and mEthanol were HPLC grade (Sigma-Aldrich, St. Louis, USA). Ultrapure water was obtained using Milli-Q (Merck, Kenilworth, USA). Reference standards included phenol (Sigma-Aldrich; 99% purity), para cresol (Sigma-Aldrich; 99% purity), urocanic acid (4-Imidazoleacrylic acid; Sigma; 99% purity), choline bitartrate (Sigma; 99% purity), L-tryptophan (Sigma; with ≥98% purity) and adenosine (Olchemim; 98% purity). Dragondorff’s reagent was used for TLC visualisation (Fluka; St. Louis, USA).
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4

Profiling Volatile Compounds in Milk

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Propionic acid, butanoic acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, n-decanoic acid, hexanal, octanal, nonanal, (E)-2-octenal, pentanol, hexanol, 1-octen-3-ol, nonanol, paracresol, 2-heptanone, 3-octanone, 2-nononone, acetophenone, ethyl butyrate, ethyl caproate, and d-limonene were purchased from Sigma-Aldrich (St. Louis, MO). These chemical substances were detected to be the odor-active compounds of milk fan Y6 sample in our previous study (Tian et al., 2019) (link). All of the chemicals were of chromatographic grade.
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5

Volatile Organic Compounds Profiling

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Propionic acid, butanoic acid, 3-methyl-butyric acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, decanoic acid , 3-methylbutyraldehyde, octanal, nonanal, (E)-2-octenal aldehyde, 1-pentanol, 1-hexanol, 1,6-octadiene-3-alcohol, nonanol, para-cresol, 2-heptanone, 3-octanone, 2-nononone, acetophenone, ethyl butyrate, ethyl caproate, ethyl octanate, and d-limonene were purchased from Sigma-Aldrich (St. Louis, MO). 2-Octanol (internal standard, IS) and n-alkane standards (C 6 -C 30 ) were purchased from Dr. Ehrenstorfer GmbH Co. Ltd., Germany. All of the chemicals were of chromatographic grade.
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