T. flavidus were purchased from Fujian Shenhai Food (Zhangzhou, China) and the skins were peeled off and minced with a meat grinder. Alcalase (EC 3.4.21.62), neutral protease (EC 3.4.22.17), and pepsin (EC 3.4.23.1) were purchased from Solarbio (Beijing, China). ACE (EC 3.4.15.1, from rabbit lung), hippuryl-L-histidyl-L-leucine (HHL), acetonitrile (ACN, HPLC grade), captopril (>99% purity), and trifluoroacetic acid (TFA) were purchased from Sigma Aldrich (St. Louis, MO, USA). Formic acid (FA) was supplied by Merck Chemical Company (Darmstadt, Germany). All other chemical reagents were of analytical grade.
Alcalase
Manufactured by Solarbio
Sourced in China
Alcalase is a proteolytic enzyme produced by the Bacillus licheniformis bacteria. It is a serine endopeptidase that can hydrolyze peptide bonds in proteins. The core function of Alcalase is to catalyze the breakdown of protein molecules into smaller peptides and amino acids.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin, by Solarbio (11 mentions)
Papain, by Solarbio (11 mentions)
Flavourzyme, by Solarbio (9 mentions)
Neutrase, by Solarbio (8 mentions)
Pepsin, by Solarbio (7 mentions)
Protamex, by Solarbio (4 mentions)
Neutral protease, by Solarbio (2 mentions)
Anhydrous ethanol, by Sinopharm (2 mentions)
Sodium hydroxide (naoh), by Sinopharm (2 mentions)
Boric acid, by Sinopharm (2 mentions)
Sephadex g 25, by Solarbio (2 mentions)
Fapgg, by Merck Group (2 mentions)
Trypsin, by Merck Group (2 mentions)
Pepsin, by Merck Group (2 mentions)
Papain, by Merck Group (2 mentions)
Trichloroacetic acid, by Sinopharm (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Solarbio (2 mentions)
Hippuryl l histidyl l leucine, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
23 protocols using alcalase
1
Enzymatic Hydrolysis of T. flavidus Skin
2
Extraction and Purification of Rice Protein
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Rice was provided by Jinjian Cereals Industry Co., Ltd. (Changde, China). The RP was obtained in the laboratory using alkaline extraction and acid precipitation (protein content: 85% ± 0.25%). Soy isoflavone was purchased from Nanjing Spring and Autumn Biological Engineering Co., Ltd. (Nanjing, China). Alcalase, neutrase, flavorzyme, and trypsin were purchased from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). All other reagents were of analytical grade and were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
3
Enzymatic Hydrolysis and Lactobacillus Taming
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Build a system in which 12% sheep bone powder was firstly hydrolyzed by 4.25% alcalase
(activity≥200 U/mg, Solarbio) at pH 9 and 45℃ for 400 min followed by adding
3.65% flavourzyme (activity≥20 U/mg, Solarbio) to the resulted mixture (adjusted to pH
6) and incubating at 55℃ for 240 min. The final mixture was autoclaved at 121℃
for 20 min. L. helveticus were gradually tamed by culturing in MRS broth with
increasing proportion of SBH (supplemented with 1% glucose) from 10% to 100%. The finally tamed
L. helveticus independent of MRS broth was used as the starter in the
following single-factor tests and responsive surface design tests.
(activity≥200 U/mg, Solarbio) at pH 9 and 45℃ for 400 min followed by adding
3.65% flavourzyme (activity≥20 U/mg, Solarbio) to the resulted mixture (adjusted to pH
6) and incubating at 55℃ for 240 min. The final mixture was autoclaved at 121℃
for 20 min. L. helveticus were gradually tamed by culturing in MRS broth with
increasing proportion of SBH (supplemented with 1% glucose) from 10% to 100%. The finally tamed
L. helveticus independent of MRS broth was used as the starter in the
following single-factor tests and responsive surface design tests.
4
Yak Bone Peptides Extraction Optimization
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Yak bones (leg bone) were obtained from the Tibet Academy of Agricultural and Animal Husbandry Sciences (Tibet, China). The bones were mechanically ground into granules with a diameter ranging from 3–5 mm and stored at −80 °C until analyzed. Pepsin (EC number 3.4.23.1, 1200 U mg−1), trypsin (EC number 3.4.21.4, 92 383 U mg−1), Alcalase (EC number 3.4.21.63, 221 756 U mg−1), Flavourzyme (EC number 3.4.11.1, 15 311 U mg−1), Protamex (56 084 U mg−1), Neutrase (EC number 3.4.24.28, 19 267 U mg−1) and papain (EC number 3.4.22.2, 37 018 U mg−1) were purchased from Solarbio (Beijing, China). Soluble peptides content was measured using a bicinchoninic acid (BCA) Protein Assay Kit from Beyotime (Shanghai, China). Trifluoroacetic acid (TFA), sodium dodecyl sulfate (SDS), and trinitrobenzene sulfonic acid (TNBS) were purchased from Sigma-Aldrich (Shanghai, China). HPLC grade acetonitrile (ACN) was purchased from Thermo Fisher Scientific (Waltham, USA). GP-16 and GD-18 (purity > 98%) was chemical synthesized and purified using high performance liquid chromatography (HPLC) by Beijing Protein Innovation (Beijing, China). All other chemicals and reagents were of analytical grade, unless otherwise stated.
5
Enzymatic Degradation of Frozen Squid
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Frozen SYC 9 ± 1 cm in length was obtained from the Zhejiang Xianghai Food Co., Ltd. in Wenzhou, China. Papain (100,000 U/g), Alcalase (100,000 U/g), Neutrase (50,000 U/g), and Xanthine oxidase (X1875-5UN, derived from bovine milk) were purchased from Solarbio Co., Ltd. (Beijing, China). We purchased 0.2 M potassium phosphate buffer (pH = 7.4) from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Xanthine (≥98%) and allopurinol (chromatographically pure) were purchased from Sigma Aldrich Co., Ltd. (St. Louis, MO, USA). Sodium hydroxide, anhydrous ethanol, and boric acid were of analytical grade and purchased from Sinopharm chemical reagent Co., Ltd. (Shanghai, China).
6
Enzymatic Hydrolysis of Antarctic Krill
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Defatted Antarctic krill powder was provided by Zhejiang Hailisheng Biotechnology Co., Ltd. (Zhoushan, China), with a proximate composition as follows: moisture (8.29%), protein (78.33%), fat (1.17%), sugar (0.84%), and ash (4.62%). Neutrase (5 × 104 U/g), Alcalase (2 × 105 U/g), Papain (1 × 105 U/g), Trypsin (2.5 × 105 U/g), Protamex (1 × 105 U/g), the BCA protein assay kit, the β-GAL assay kit, and the LDH assay kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). L. rhamnosus ATCC7469 was provided by Shanghai Luwei Microbial Sci. &Tech. Co., Ltd. (Shanghai, China). MRS broth and MRS agar were purchased from Qingdao Hope Bio-Technology Co., Ltd. (Tsingtao, China). The other reagents and chemicals were of analytical grade.
7
Extraction and Characterization of EBN
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Indonesia’s 006 factories provided EBN by-products, which were identified as genuine by Lin Jiang, an associate professor of Sun Yat-Sen University. Six proteases (Papain, acid protease, neutrase, alcalase, pepsin, and trypsin), superoxide anion kits, and Sephadex G-25 were obtained from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). 2,2-diphenyl-1-picrylhydrazyl (DPPH), O-phthaldialdehyde (OPA), ascorbic acid (VIT C), and HPLC-grade trifluoroacetic acid (TFA) were acquired from Macklin (Shanghai, China). 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid (ABTS) kits were obtained from Beyotime Bio-Technology Co., Ltd. (Shanghai, China). Serine standard was acquired from Yuanye Biotechnology Co., Ltd. (Shanghai, China). HPLC-grade acetonitrile was obtained from Oceanpak (Stockholm, Sweden). Edaravone was acquired from Aladdin (Shanghai, China). Sodium nitroprusside (SNP) was acquired from Sigma-Aldrich Co. (St. Louis, MO, USA). Other reagents and solvents were of analytical grades.
8
Characterization of Mealworm Enzymatic Activities
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Fresh T. molitor were donated by Prof. Ning Li at Chongqing Technology and Business University, while 8-anilino-1-naphthalenesulfonic acid (ANS), trypsin (250 N.F.U/mg), flavourzyme (≥30,000 U/g), papain (≥800,000 U/g), and alcalase (≥200,000 U/g) were bought from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). The o-phthaldialdehyde (OPA) was bought from Shanghai Bide Medical Technology Co., Ltd. (Shanghai, China). The acetonitrile, trifluoroacetic acid, and formic acid were chromatographic grade and were supplied by Chengdu Kelong Chemical Co., Ltd. (Chengdu, China). DPP-4 (≥4000 units/μg) and its substrate Gly-Pro-p-nitroanilide were bought from Millipore Sigma (Burlington, MA, USA).
9
Enzymatic Extraction of Purple Wheat Protein
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Purple wheat (Triticum aestivum L.) of the cultivar Shannong Purple Wheat One were harvested in August, 2018 (base on the GB 1351–2008) at maturity from Shangdong, China. Purple wheat bran protein (PWBP) was prepared from purple wheat bran. Alcalase (2 × 105 U/g), flavourzyme (3 × 104 U/g), papain (8 × 105 U/g), neutral enzyme (3 × 104 U/g), and trypsin (2.5 × 105 U/g) were purchased from Beijing Solarbio Science &Technology Co., Ltd. (Beijing, China) DETA-Sepharose FF and Sephadex G-25 were purchased from Beijing Solarbio Science &Technology Co., Ltd. (Beijing, China).
10
Antioxidant and ACE Inhibitory Effects of Skipjack Tuna Roe
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Skipjack tuna roes were provided by Ningbo Today Food Co., Ltd. (Zhejiang, China). HUVECs were purchased from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). ROS (Product no. E004-1-1), Hoechst33342 (Product no. G023-1-1), CAT (Product no. A007-1-1), SOD (Product no. A001-3-2), GSH-PX (Product no. A005-1-2), MDA (Product no. A003-1-2), ET-1 (Product no. H093-1-2), and NO (Product no. A013-2-1) kits were purchased from Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). Glutathione (GSH), trypsin, pepsin, papain, ACE and FAPGG were purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd. (China). Alcalase, MTT, DMEM, DMSO, fetal bovine serum (FBS), norepinephrine (NE), trifluoro acetic acid (TFA), neutrase and Cap were purchased from Beijing Solarbio Science & Technology Co., Ltd. (China). ACEi peptides of WGESF (TRP3), IKSW (TRP6), YSHM (TRP9), and WSPGF (TRP12) with purity higher than 95% were synthesized in Shanghai Apeptide Co. Ltd. (China).
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