Structural Changes,Volatile Compounds and Antioxidant Activities of Maillard Reaction Products Derived from Scallop (Patinopecten yessoensis) Female Gonad Hydrolysates

In order to improve the utilization of scallop byproducts, scallop female gonad hydrolysates (SFGHs) were prepared using alcalase and ribose at a mass ratio of 1:2 and pH 7.0 (95°C, up to 12 h) to develop heterogeneous Maillard-type antioxidant and flavor compounds. The formation of Schiff’s base as well as modification of the amide bands in Maillard reaction products (MRPs) were investigated using ultraviolet–visible, fluorescence, and Fourier transform infrared spectroscopy. The significant decrease of amino acid residues explained the strong Maillard reactivity of SFGHs. MRPs with enhanced ABTS⁺ radical scavenging capacity over SFGHs promoted the survival of HepG2 cells treated with hydrogen peroxide. Moreover, 19 new volatile compounds were produced by the reaction of SFGHs with ribose. These results suggest that SFGHs–ribose MRPs can be potentially used as antioxidants with flavor properties.

Abbreviations: SFGs: scallop female gonads; SFGHs: scallop female gonad hydrolysates; ABTS: 2?2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid); BHT: butylated hydroxytoluene; Trolox: 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; MRPs: Maillard reaction product; H₂O₂: hydrogen peroxide; HS-SPME/GC/MS: Headspace solid phase micro-extraction/gas chromatography/mass spectrometry.     

Recovery of Proteins from Squid By-Products with Enzymatic Hydrolysis and Increasing the Hydrolysate’s Bioactivity by Maillard Reaction

ABSTRACT

The Maillard reaction is an effective method for enhancing the activity of bioactive peptides. In this study, squid by-product hydrolytic peptides (SPHPs) were obtained. The effects of the Maillard reaction on the antioxidant and antibacterial activities of the SPHPs were explored. The results showed most of the peptides in the < 500 Da fraction of SPHPs reacted with D-arabinose to form the 500–1000 Da fraction (89.60%). The Maillard reaction products (MRPs) possessed more potent antioxidant activities, including free radical scavenging activity, ferrous chelating capacity, and reducing power, than the SPHPs. Although both the SPHPs and MRPs had broad-spectrum antibiotic activities and could inhibit the growth of both gram-negative and gram-positive bacteria, the antibacterial activities of the MRPs were higher than those of the SPHPs. Furthermore, the efficacy of the MRPs lasted significantly longer than that of ampicillin and could last for more than 30 days. In addition, both the SPHPs and MRPs had good functional properties and many potential applications. The results suggested that the Maillard reaction has potential to be used to improve the activities and physicochemical properties of bioactive peptides.     

Effects of Freshness and Heating Temperature on the Discoloration of Squid Meat Products

ABSTRACT

Squid processed products such as dried-seasoned squid turn brown during storage, causing a change in quality. The main cause of this browning is melanoidin produced by the Maillard reaction. In this study, four types of squid were processed at different heating temperatures between 50 and 90°C and examined for suppression of the production of D-ribose, the main component of melanoidin. Furthermore, the color change of the squid meat with each heat treatment during storage of 32 days at 30°C was investigated. Bigfin reef squid and cuttlefish with a relatively high freshness decreased D-ribose generation as the heating temperature increased, and the browning during storage also decreased accordingly. In contrast, Humboldt squid with a low freshness did not generate new D-ribose at any heating temperature during storage. Nevertheless, the browning was reduced as the heating temperature increased. It is thought that the Maillard reaction was also reduced by the heat denaturation of muscular proteins. These results suggest that browning due to the Maillard reaction of squid meat is greatly affected by initial freshness and heating temperature.     

Production of Antioxidative Maillard Reaction Products from Gelatin Hydrolysate of Unicorn Leatherjacket Skin

Antioxidative activities of Maillard reaction products (MRPs) derived from gelatin hydrolysate (GH) of unicorn leatherjacket skin as affected by types of saccharides, GH to saccharide ratio, incubation temperatures, relative humidity (RH), and times were investigated. MRPs prepared using mixture of GH and galactose at 2:1 (w/w) ratio showed the highest antioxidative activity. At different incubation temperatures (50–70°C) and RHs (55–75%), optimal condition for preparing antioxidative MRPs included heating GH and galactose (2:1) powder at 70°C and 55% RH for 36 h. Electrophoresis and Fourier transform infrared spectroscopy revealed the formation of high molecular weight polymers, and the advanced glycation took place in MRPs.     

Optimization of Antioxidant Peptides Production from the Mantle of Cuttlefish (Sepia pharaonis) Using RSM and Fractionation

Persian Gulf cuttlefish mantles were hydrolyzed (CPH) using alcalase, and the optimal hydrolysis parameters were obtained for the highest degree of hydrolysis (DH) and strongest antioxidant (based on their ability to quench 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals) activity using response surface methodology (RSM). The predicted optimal parameters of DH and quenching DPPH radicals was: pH of 7.88, 50.2°C, 150 min, and enzyme to substrate ratio of 1.5%. The reducing power (RP) and ability of optimized peptides to quench ABTS radicals in a gastro-intestinal track model system increased during the intestinal stage, while scavenging ability against DPPH radicals dropped (P < 0.05). The oxidation of lipid was retarded in a lecithin-liposome model added with optimized CPH in a concentration dependent response. Ultrafiltration of optimized CPH showed that the 3–10 KDa fraction had the greatest DPPH radical scavenging activity, the 10–30 KDa fraction had the highest reducing power, and the <3 KDa fraction had the greatest ABTS radical scavenging activity.     

Heat- and light-treatment change the visible fluorescence of Akoya cultured pearls

We exposed Akoya cultured pearls separately to heat (60–120 °C) and artificial light to investigate changes to fluorescence in the visible range and yellowing. We found that for both heat-treated and light-treated pearls, the fluorescence peak shifted from 480 to 430 nm with an increase in fluorescence intensity. This change in intensity was more prominent in heat-treated pearls, with the initial speed of increase rising with treatment temperature; treatment at 100 °C caused the greatest increase in fluorescence intensity. However, aminoguanidine suppressed the heat-induced change in the fluorescence of an ethylenediaminetetraacetic acid–soluble nacreous layer matrix. These results suggest that the heated-induced changes in the fluorescence of Akoya cultured pearls were caused largely by a buildup of fluorescent advanced glycation end products through the Maillard reaction. Although heat treatment led to a large increase in fluorescence intensity of the peak at approximately 430 nm in a deoxygenized environment, hardly any change in fluorescence intensity was observed after light treatment in this environment. Moreover, a new shoulder peak appeared at about 460 nm after light treatment. These results suggest that the Maillard reaction was not a major factor in the light-induced changes in the fluorescence of Akoya cultured pearls.     

Characterization of Flavor Properties from Fish (Collichthys niveatus) Through Enzymatic Hydrolysis and the Maillard Reaction

ABSTRACT

Collichthys niveatus (C. niveatus) is an abundant but underutilized fish species caught in China. In this study, flavor properties of the Maillard reaction products (MRPs) obtained from C. niveatus protein hydrolysates were determined. The optimized fish protein hydrolysates were prepared using commercial Alcalase, and the optimum values of temperature, pH, and enzyme/substrate ratio were 55.6°C, 8.7, and 1.03%, respectively. In addition, the volatile compounds of the MRPs were evaluated using headspace solid–phase microextraction combined with gas chromatography mass spectrometry. A total of 80 volatile compounds were separated and identified. Priazines, alcohols, and aldehydes were the major flavor contributors to the aroma of the MRPs (accounting for 38.86, 9.21, and 8.23%, respectively), while pyrazines were the most abundant group of volatile compounds present, with 2,5-dimethyl-pyrazine, methylpyrazine, and 3-ethyl-2,5-dimethylpyrazine being found in highest quantities (19.83, 5.81, and 5.70%, respectively). Other components—including alcohols, aldehydes, furans, ketones, and esters—were also identified in the MRPs. This study provides a systematic strategy for marine products processing and volatiles characterization. The results point to the information in the volatiles from the MRPs of protein hydrolysis from C. niveatus and pave the way for full utilization of this species.     

Chemical Characterization,Temperature Stability,and Enzymatic Studies on Edible Marine Algae Kappaphycus alvarezii (Doty)

Antioxidant activity of the marine alga Kappaphycus alvarezii was investigated. Methanol, acetone, petroleum ether, aqueous methanol, ethyl acetate, and chloroform extracts (1 mg/mL) of K. alvarezii were tested for their 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. The values were compared with those of Vitamin C and butylated hydroxytoluene (BHT). Extracts showing positive results, when tested for DPPH free radical scavenging, were examined for dose effect, in-vivo hydrogen peroxide (H₂O₂) and hydroxyl radical scavenging assays. All extracts showed dose-dependent DPPH scavenging and significant hydroxyl radical scavenging activities (> 82.6%). The acetone, aqueous methanol, and methanol extracts of K. alvarezii showed the highest scavenging activity. Ethyl acetate extract showed a moderate activity of 62.9%. In the DPPH method, petroleum ether and hexane extracts showed less activity with IC₅₀ values of 118.58 ± 8.94 and 116.25 ± 7.14 μg/mL, respectively. Acetone, methanol, and ethyl acetate extracts exhibited IC₅₀ values of 57.32 ± 1.07, 61.31 ± 0.67, and 79.50 ± 1.59 μg/mL, respectively. K. alvarezii showed higher antioxidant activity in both in vitro and in vivo studies. Proton NMR studies revealed signals in the region 0.5 to 2.0 ppm suggesting the presence of steroidal identity in the extracts.     

Evaluation of Flavor Improvement in Antarctic Krill Defluoridated Hydrolysate by Maillard Reaction Using Sensory Analysis,E-nose,and GC-MS

ABSTRACT

Antarctic krill is a high-quality animal protein resource, but the presence of fluorine limits its development. Antarctic krill defluoridated hydrolysate products (DHPs) generally have a bitter taste that is unacceptable to consumers. Therefore, it is necessary to improve the flavor of defluoridated hydrolysate. In this work, flavor improvement of DHPs by the Maillard reaction was evaluated by sensory analysis, electronic nose (E-nose), and gas chromatography-mass spectrometry (GC-MS). The potential correlations among the signals of sensory attributes, E-nose, and volatile components were analyzed using the analysis of partial least squares regression (PLSR). A total of 63 volatile compounds were identified and quantified via GC-MS. The flavors in Maillard reaction products were mainly attributed to pyrazines, aldehydes, ketones, and furans. After the Maillard reaction, the two main aromatic compounds generated were 3-ethyl-2,5-dimethylpyrazin (roast odor, OAV = 32.7) and 3-(methylthio) propionaldehyde (shrimp meat odor, OAV = 16.43), which contributed to flavor improvement in DHPs. PLSR results showed a good correlation between most variables of sensory attributes, E-nose, and volatile compounds, which indicates that the overall flavor of DHPs was improved by the Maillard reaction.

Abbreviations: DHPs: defluoridated hydrolysate products; MRPs: Maillard reaction products; GC-MS: gas chromatography-mass spectrometry; E-nose: electronic nose; OAV: odor activity value; SPME: solid-phase microextraction; GC-O: gas chromatography-olfactometry; PCA: principal component analysis; PLSR: partial least squares regression     

Feasibility study on water solubilization of spawned out salmon meat by conjugation with alginate oligosaccharide

A feasibility study was carried out to determine whether water-soluble salmon meat could be manufactured by conjugating a glycosyl unit using the Maillard reaction. Spawned out salmon meat was washed, mixed with alginate oligosaccharide (AO) and sorbitol, lyophilized, and then heated at 60°C and 5–95% relative humidity (RH) to introduce AO (the mean degree of polymerization was six) into the myofibrillar proteins through the Maillard reaction. The reaction progressed with an increase in the reaction humidity and the amount of AO bound to the protein reached >150 μg/mg at RH 65 and 90%. However, the protein glycosylation under high humidity impaired protein solubility and the meat protein became effectively water-soluble with the conjugation with AO at reaction conditions of 60°C and RH 35%. The improved characteristics of the meat protein were highly stable at room temperature. Further, the water-soluble protein can be prepared from the frozen salmon meat stored at −25°C for 60–90 days. These results indicate that protein glycosylation has strong potential for use with spawned out chum salmon. The suppression of protein denaturation during processing is important to obtain the high water-soluble meat protein.     

4种海藻膳食纤维清除自由基的比较研究

采用羟基自由基体系、超氧阴离子自由基体系、烷基自由基引发的亚油酸氧化体系和DPPH体系对江蓠(Gracilaria)、麒麟菜(Eucheuma)、马尾藻(Sargassum)、海带(Laminaria)4种膳食纤维清除自由基的活性进行研究。结果表明，在羟基自由基体系中，江蓠、麒麟菜、马尾藻、海带4种膳食纤维的IC50分别为5．2mg／mL、5．5mg／mL、4．4mg／mL和4．1mg/mL；在超氧阴离子自由基体系中，江蓠、麒麟菜膳食纤维的IC50分别为4．8mg/mL和5．3mg/mL，而马尾藻、海带膳食纤维的最高清除率分别为36％和42％；在烷基自由基引发的亚油酸氧化体系中，江蓠、麒麟菜膳食纤维的最高清除率分别为36％和26％，马尾藻、海带膳食纤维的IC50分别为4．1mg/mL和3．5mg/mL；在DlPIH体系中，江蓠、麒麟菜、马尾藻膳食纤维的最高清除率分别为31％、11％和26％，海带膳食纤维的IC50为6．2mg/mL。在各体系中，与相应的专一性阳性参照物对比，证明这4种海藻膳食纤维对自由基有一定的清除作用。     

Physicochemical and Reactive Oxygen Species Scavenging Properties of Collagen and Collagen Hydrolysates from Farmed Globefish (Fugu rubripes) Bone

Globefish (Fugu rubripes) bone collagen (GBC) was isolated and characterized. Electrophoresis and Fourier transform infrared (FTIR) spectra indicated that GBC was high-purity type I collagen with an intact triple-helical structure. Amino acid analysis revealed that GBC contained higher glycine content (364 residues/1,000 residues) than that of other fishery bones, and the imino acid content was 189 residues/1,000 residues. The denaturation temperature (T_d) of GBC was 27.1°C. GBC displayed a rapid fibril-forming rate and well-defined fibril morphology in vitro. Globefish bone collagen hydrolysates (GBCH) were optimized by neutral protease through response surface methodology (RSM) with the highest hydroxyl radical (?OH) scavenging capacity. The optimal hydrolysis condition was a reaction temperature of 48.4°C, time of 120 min, and enzyme/substrate ratio of 2520 U/g. Three fractions were collected by ultrafiltration, and the fraction GBCH-III (Mw<1 kDa) possessed the strongest ?OH scavenging ability. Reactive oxygen species (ROS) scavenging capacities were evaluated with two model systems: ?OH and nitric oxide (NO?). The ROS scavenging capacities increased in a dose-dependent manner. GBCH-III proved to be a more effective ?OH scavenger than reduced glutathione (GSH). These findings demonstrated that GBC and GBCH have potential as an alternative source of collagen-derived materials for use in pharmaceutical, nutraceutical, and cosmetic industries.     

鲣鳔蛋白抗氧化酶解物制备工艺

为有效提高鲣鳔蛋白的附加值,研究以DPPH自由基清除率为抗氧化活性评价指标,采用蛋白酶酶解制备活性多肽的工艺,选用菠萝蛋白酶、复合蛋白酶、碱性蛋白酶、木瓜蛋白酶、胃蛋白酶、胰蛋白酶、中性蛋白酶7种酶在各自最适的条件下酶解,筛选出复合蛋白酶为最适用酶,通过单因素实验分别研究加酶量、溶液初始p H、酶解温度和时间对酶解物抗氧化活性的影响,在此基础上,根据响应面法优化鲣鳔抗氧化酶解物的制备工艺。结果显示,最佳酶解工艺条件为加酶量8.53 U/mg,p H 5.54,温度50.03°C,时间5.07 h。此外,利用超滤法对最佳条件下制备的酶解物进行初步分级,得到分子质量分别为大于10 000 u、3000~10 000 u和小于3000 u的3段组分,且这3段组分对DPPH自由基的半抑制浓度IC50值分别为0.64、0.52和0.37 mg/m L。研究表明,最优条件下制备的酶解物的DPPH清除率达72.00%,与模型预测值71.60%接近,且其中小于3000 u的组分具有较强的DPPH自由基清除活性。     

Quality Changes and Browning Developments During Storage of Dried-Seasoned Squid (Dosidicus gigas and Ommastrephes bartrami)

ABSTRACT

The physicochemical changes—moisture, water activity, crude protein, total volatile basic nitrogen (TVB-N), trimethylamine oxide (TMAO), dimethylamine (DMA) and formaldehyde, reducing sugar and amino acid; microbiological changes—aerobic plate count (APC) and coliforms; and browning development (browning intensity and color) in dried-seasoned squids (Dosidicus gigas and Ommastrephes bartrami) during storage at 25°C were investigated. After the storage, the moisture and water activity decreased significantly. The content of crude protein decreased, accompanied by progressive accumulation of TVB-N and slow increase of APC. The high level of TMAO in dried-seasoned squid gradually decreased, whereas DMA and formaldehyde increased significantly during the whole period of storage. The browning intensity and b* value significantly increased, which revealed the occurrence of browning in two squid products. The dominant reducing sugar, lactose, and glucose, and the total amino acid including lysine and proline, decreased remarkably, which were well-correlated with the browning development. The quality deterioration in jumbo squid was more rapid than in neon flying squid partially due to the high level of reducing sugar in the former product. To improve quality and resist browning reaction of dried squid product during storage, the addition of extrinsic reducing sugar became a control factor.     

1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging by protein hydrolyzates from tuna cooking juice

ABSTRACT: Protease XXIII, from Aspergillus oryzae , was used to hydrolyze tuna cooking juice at 37°C for up to 6 h. The hydrolyzate obtained at the degree of hydrolysis of 25.68% (after hydrolysis for 2.5 h) displayed the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging effect, reaching 82.19%. Six major fractions (A, B, C, D, E, and F) of this hydrolyzate were obtained by Sephadex G-25 column chromatography using a 0.05 M phosphate buffer (pH 6.5) as the mobile phase. All six fractions displayed a scavenging effect for the DPPH radical, but the scavenging effect was only obvious in two fractions (B and C). After the solid content of hydrolyzates was concentrated from one to five times, the scavenging effect of the DPPH radical increased from 17% to 75% for fraction B, and from 13% to 66% for fraction C. Seven anti-oxidative peptides were isolated from the hydrolyzates (mixture of B and C fractions) by reversed-phase HPLC. The peptide sequences comprised four to eight amino acid residues, including Val, Ser, Pro, His, Ala, Asp, Lys, Glu, Gly, or Tyr.     

二种海胆性腺的脂质组成及其抗氧化活性

为分析马粪海胆和光棘球海胆性腺的脂质组成和抗氧化活性,采用核磁共振和气相色谱—质谱技术对2种海胆性腺油脂的脂质成分和脂肪酸组成进行分析,并通过DPPH自由基清除法、羟基自由基清除法和超氧阴离子自由基清除法对其脂质的抗氧化活性进行研究。结果显示,马粪海胆和光棘球海胆性腺脂质均以甘油三酯和磷脂为主,胆固醇、胆固醇酯和游离脂肪酸含量较低。马粪海胆和光棘球海胆性腺总脂富含C20:4n-6和C20:5n-3,且二者总量分别占脂肪酸含量的35.88%和34.98%;同时2种海胆性腺的中性脂和极性脂的脂肪酸组成存在较大差异,中性脂以C14:0和C16:0等饱和脂肪酸为主,而极性脂以C20:4n-6和C20:5n-3等多不饱和脂肪酸为主。马粪海胆和光棘球海胆性腺脂质对DPPH自由基、羟基自由基和超氧阴离子自由基均具有较好的清除能力,DPPH自由基IC₅₀分别为2.75和1.98 mg/mL,羟基自由基IC₅₀分别为0.33和0.29 mg/mL,超氧阴离子自由基IC₅₀分别为0.33和0.31 mg/mL。研究表明,马粪海胆和光棘球海胆性腺脂质具有较高的营养价值和抗氧化活性,可作为C20:4n-6、C20:5n-3和磷脂等功能性脂质因子的重要膳食来源。     

Application of Two-Level Full Factorial Design for the Extraction of Fucoxanthin and Antioxidant Activities from Sargassum siliquosum and Sargassum polycystum

Two-level full factorial design was employed to identify the extraction parameters that can improve the derivation of fucoxanthin content (FC), total carotenoid content (TCC), and antioxidant from two brown seaweeds, Sargassum siliquosum (SS) and Sargassum polycystum (SP). These parameters included temperature (A: 4–45°C), time (B: 30–1,440 min), and solvent-to-solid ratio (C: 10–50 ml/g). Antioxidant activities were determined as trolox equivalent antioxidant capacity (TEAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, and ferric-reducing antioxidant power (FRAP). Results showed that all three factors were significant (p < 0.05) in providing higher FC in both species. These factors were also significant in obtaining higher TCC in SS; whereas in SP, TCC was only affected by solvent-to-solid ratio. Only temperature was found to contribute significantly to a higher TEAC in both species. However, none of the factors improved DPPH for SS, except temperature and time for SP. For SS, only time was significant in obtaining higher FRAP; whereas temperature and time were significant for SP. Hence, results indicate that a simple modification in the extraction temperature, time, and solvent-to-solid ratio will be able to improve the derivation of fucoxanthin, carotenoids, and antioxidant activities.     

Protein Hydrolysates and Ultrafiltration Fractions Obtained from Krill (Euphausia superba): Nutritional,Functional, Antioxidant,and ACE-Inhibitory Characterization

ABSTRACT

Krill (Euphausia superba) was hydrolyzed by proteolytic enzymes in order to produce multifunctional bioactive peptides, and their functional properties were evaluated. Krill protein hydrolysate (KPH) by pepsin with 4-h hydrolysis showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and angiotensin I converting enzyme (ACE) inhibitory activities. The solubility and foaming properties of KPH were higher than those of the unhydrolyzed krill protein at a wide range of pHs. KPH was further fractionated based on molecular weight. The 1- to 3-kDa peptide fraction exhibited the highest DPPH scavenging activity (IC₅₀ value of 0.5 mg/mL), oxygen radical absorbance capacity (497.39 ± 4.31 µM TE/mg fraction), 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid cation radical scavenging activity (48.41 ± 0.23 µM TE/mg fraction), and reducing power (110.40 ± 2.07 µM TE/mg fraction). However, the < 1-kDa peptide fraction exhibited a higher ACE inhibitory activity than that of other fractions. The 1- to 3- and < 1-kDa peptide fractions are rich in aromatic and hydrophobic amino acids, respectively.     

Free radical scavenging ability and structure of a 90-kDa protein from the scallop shell

We have previously reported that the scallop shell extract possesses free radical scavenging activity. In this study, we isolated a glycoprotein with a molecular weight of approximately 90 kDa (named 90-kDa protein) which showed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide anion radical scavenging, reducing, and ferrous ion-chelating activities. Amino acid composition analysis showed that the 90-kDa protein was rich in Asx (Asp or Asn), Ser and Gly residues. A BLAST search of partial amino acid sequences determined by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry revealed that the 90-kDa protein is a novel protein. The 90-kDa protein is a yellow protein that contains fluorescent substances. To the best of our knowledge, this is the first report of a radical scavenging protein from the scallop shell.     

Conditions for the induction of some selective enzymes from <Emphasis Type="Italic">Bacillus subtilis</Emphasis> and their hydrolysis ability against mackerel and gracilar (asparagus, <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis>)

ABSTRACT:   This study was conducted to optimize the cultivation conditions for Bacillus subtilis to produce proteases, amylases and cellulase, and to further investigate the hydrolysis ability against mackerel and asparagus. The extracellular enzymes from B. subtilis after 2 and 4 days incubation in a modified medium, containing 1% skim milk, 1% soya meal, 0.25% starch, 0.25% K₂HPO₄, 0.5% NaCl and 0.05% MgCl₂ were collected for the hydrolysis of asparagus and minced mackerel, respectively. Except for the α,α-diphenyl-β-picrylhydrazyl (DPPH) scavenging ability of the hydrolyzed asparagus, the trolox equivalent antioxidation capacity and DPPH scavenging ability of both samples increased significantly ( P  < 0.05) after 1 h hydrolysis and further increased during elongated hydrolysis at 50°C. Sodium dodecylsulfate–polyacrylamide gel electrophoresis indicated severe degradation of muscle proteins during hydrolysis. Changes in reducing sugar, soluble proteins and peptides before/after hydrolysis suggested the extracellular enzymes from B. subtilis could effectively hydrolyze the mackerel or asparagus, and subsequently improve their antioxidation ability.