Glycerol

M. smegmatis was grown in nutritious Middlebrook 7H9 supplemented with 0.5% (v/v) glycerol (Sangon Biotech, Shanghai), 0.05% (v/v) Tween-80 (Sangon Biotech, Shanghai) and 10% (v/v) ADC. MICs were quantified with the resazurin microdilution titration method. Briefly, Middlebrook 7H9 broth was added to 96-well plates to prepare a continuous two fold dilution of rifampicin in triplicate. Each well was inoculated with 10⁵ CFU bacteria for 2–3 days at 37 °C and subsequently mixed with 30 μl 0.01% resazurin (Sangon Biotech, Shanghai) solution to determine the lowest concentration that inhibited bacterial growth. Three different gradients of rifampicin were tested: 600, 300, 150, 75, 37.5, and 18.75 μg/ml; 500, 250, 125, 62.5, 31.25, and 15.625 μg/ml; and 400, 200, 100, 50, 25, and 12.5 μg/ml.

Aeromonas hydrophila strains AH33, AH189, AH196, and AH301 (Table 1) were isolated from diseased carps and identified based on gyrB sequences. Strain NJ‐35 was donated by Prof. Yongjie Liu (College of Veterinary Medicine, Nanjing Agricultural University, China) (Pang et al., 2015). Stock cultures were maintained at −80°C in Luria‐Bertani broth (Oxoid, Basingstoke, UK) containing 30% (v/v) glycerol (Sangon Biotech, Shanghai, China). When required, the stocks were streaked on nutrient agar, incubated at 30°C overnight, and single colonies were collected and used in subsequent experiments.
The catecholamine hormone NE (noradrenaline bitartrate) was purchased from Target Molecule (Boston). Before each experiment, NE solutions were freshly prepared with sterilized physiological saline solution and filter‐sterilized using 0.22 μm MCE syringe filters (Sangon Biotech, Shanghai, China).
Serum‐SAPI medium was prepared as described by Lyte and Ernst (1992) and Dong et al. (2016) with slight modification. Briefly, the medium contained 0.4990 g glucose, 0.5003 g NH₄NO₃, 0.2504 g KH₂PO₄, 0.2497 g KCl, and 0.1216 g MgSO₄ in one liter of 10 mM HEPES buffer, which was supplemented with 10% (v/v) fetal bovine serum (FBS, Zhejiang Tianhang Biotechnology, Hangzhou, China).

Yeast extract and tryptone were purchased from Oxoid (Shanghai, China). Pyridoxal-5’-phosphate (PLP), pyruvate and R-1-phenylethylamine (1-(R)-PEA) were purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Choline chloride (ChCl), urea, ethylene glycol (EG), propylene glycol (PG), glycerol, formate, kanamycin sulphate, isopropyl–β-d-thiogalactopyranoside (IPTG), sodium dihydrogen phosphate, disodium phosphate, imidazole, sodium chloride, Ni-NTA Sefinose™ Resin (Settled Resin), Modified Bradford Protein Assay kit and SDS PAGE gel kit were obtained from Sangon (Shanghai, China). Amicon^®® Ultra Filter units were obtained from Sigma Aldrich (Shanghai, China). All chemicals were of analytical grade. The transaminase genes from Aspergillus terreus, which were previously transformed in E. coli BL21 (DE3) pRSF-D-AAO competent cells, were stored in our laboratory at −80 °C. The plasmids and the E. coli BL21 (DE3) pRSF-D-AAO competent cells were constructed by Anhui General Biology Company (Anhui, China).

Arachidonic acid (AA), dimethyl sulfoxide (DMSO), and o-dianisdine were purchased from Sigma-Aldrich Inc. (St. Louis, MO, USA). The salvianolic acid B (SaB), danshensu sodium (DSS), lithospermic acid (LA), rosmarinic acid (RA), p-coumaric acid (pCA), and hydroxysafflor yellow A (HSYA) were purchased from Shanghai Winherb Medical Technology Co., Ltd. (Shanghai, China). The aspirin was obtained from Dalian Meilun Biological Technology Co., Ltd. (Dalian, China). The 10 batches of normal danhong injection samples, the danshen component (DS), and the honghua component (HH) were provided by Danhong Pharmaceutical Co., Ltd. (Heze, China), while five batches of abnormal samples were collected from samples that suffered oxidative degradation. The glycerol and the 4% paraformaldehyde fix solution were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Ultrahigh-purity water was produced by a Millipore Milli-Q System (Milford, MA, USA). Acetonitrile, formic acid, and methanol were obtained from Merck KGaA (HPLC grade, Darmstadt, Germany). 1-phenyl-2-thiourea was purchased from Sigma-Aldrich Inc. (St. Louis, MO, USA).

One gram of sour porridge was put into a conical flask containing 9 mL of sterile sodium chloride (0.9%, w/v) covered with glass beads. Then the slurry was shocked for 30 min in a shaker at room temperature, followed by centrifugation at 2500× g for 15 min. The supernatant was collected, serially diluted, and 1 mL of the suspension from 10⁶ dilution was plated onto the sterilized Man Rogosa Sharpe (MRS) [38 (link)] agar medium (Sangon). The plates were cultured at 30 °C for 1–2 days. Individual colonies were selected and purified. The morphological and physicochemical properties of all pure isolates were analyzed using Gram staining test, endospore test, cellular morphology and colony characteristics, biochemical tests (i.e., catalase, oxidase, nitrate reductase, arginine, amygdalin, etc.), the potentiality to ferment various saccharides such as sucrose, mannitol, glucose, arabinose and so on. All isolates were maintained at −80 °C in MRS broth supplemented with 20% (v/v) glycerol (Sangon).