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4 protocols using 2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid

1

Adlay Fermentation Protocol

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Polished adlay used in this study was provided by Guizhou Renxin Agricultural Development Co., Ltd., and was sealed in plastic bags and stored at 4°C until use. B. subtilis BJ3‐2 was procured from Dr. Wu (College of life sciences, Guizhou University). BPA was prepared as described by Wen et al. (2019). Tetramethylpyrazine, acetoin, γ‐aminobutyric acid, oleanolic acid, gallic acid, rutin, 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS), and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) were purchased from Beijing Solarbio Science & Technology co., Ltd. (Beijing, China). All other chemicals were of analytical grade.
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2

Surimi-Based Antioxidant Evaluation

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Frozen silver carp surimi was purchased from Fujian Anjing Foods Co., Ltd. (Xiamen, China), and kept at −20 °C until used. Black tea powder (caffeine removed) was purchased from Damin Foodstuff (Zhangzhou) Co., Ltd. (Zhangzhou, China). Tapioca starch, phosphate, salt and glutamine transaminase (TG enzyme) were commercially available.
1,1-diphenyl-2-picrylhydrazyl (DPPH) was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China); 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was purchased from Beijing Solarbio Technology Co., Ltd. (Beijing, China); sodium dihydrogen phosphate, dibasic sodium phosphate, trichloroacetic acid, hydrogen peroxide, ferrous sulfate and potassium persulfate were of analytical grade and purchased from Xilong Chemical Co., Ltd. (Guangzhou, China). Absolute ethanol was purchased from Sinopharm Group Chemical Reagent Co., Ltd. (Shanghai, China); ferric chloride, potassium ferricyanide, and 3,5-Dinitrosalicylic acid were purchased from Shanghai Macklin Biochemical Technology Co., Ltd. (Shanghai, China).
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Antioxidant Activity Assay Protocol

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Lecithin (purity > 98 %) and zein (purity > 98 %) were purchased from Macklin (Shanghai, China). Astilbin (purity > 98 %) was purchased from Kangbang (Chengdu, China). 30 % Hydrogen peroxide was purchased from Jingshan (Chengdu, China). Salicylic acid, KCl, NaCl, acetic acid, ethanol, ascorbic acid, and macroporous resin D101 were purchased from Kelong (Chengdu, China). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was purchased from Basifu (Hefei, China). 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was purchased from Solarbio (Beijing, China). Penicillin-streptomycin was purchased from Hyclone (Logan, UT, USA). Dulbecco’s modified essential medium (DMEM, high glucose), phosphate buffered saline (PBS), and trypsin digest were purchased from Servicebio (Wuhan, China). HepG2 cells and fetal bovine serum were provided by Procell (Wuhan, China). The reactive oxygen species detection assay kit was purchased from Biyuntian (Shanghai, China). Dimethyl sulfoxide (DMSO‑d6, 99.9 %) were purchased from Siyituopu (Wuhan, China). Ultrapure water was obtained using the Milli-Q system (Milli-pore, USA). All other reagents used were of analytical grade.
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Antimicrobial and Antioxidant Assay Protocols

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2,4,6-tripyridyl-s-trizine (TPTZ), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2′- azobis [2-methylpropanamidine] dihydrochloride (AAPH), 1,1-diphenyl-2-picrylhydrazyl (DPPH), sodium ascorbate (VC), and Tween-80: Sabouraud Dextrose Aga reagents were purchased from Solarbio (Beijing, China). Sodium acetate (trihydrate), ferrous sulfide (hexahydrate), glacial acetic acid, and 2,6-di-tert-butyl-p-cresol (BHT) were purchased from Xilong Science Co., Ltd. (Shenzhen, China). Penicillin G sodium salt (PGSS) was purchased from Zhongkeruitai Biotechnology Co., Ltd. (Beijing, China); Escherichia coli (ATCC25922; E. coli), Staphylococcus aureus (ATCC27217; S. aureus), and Candida albicans (ATCC10231; C. albicans) were purchased from Jiangsu Edison Biotechnology Co., Ltd. (Yancheng, China). Luria–Bertani culture medium was prepared by ourselves.
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