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.     

Antioxidant Activity of Brown Seaweeds

Antioxidant potential of three brown seaweeds, Anthophycus longifolius, Sargassum plagiophyllum, and Sargassum myriocystum, obtained from the Gulf of Mannar region of India were evaluated utilizing different in vitro systems. Ethyl acetate (EtOAc) fraction of Anthophycus longifolius registered significantly greater hydroxyl radical scavenging ability (IC₅₀ 0.19 mg/mL) and was effective in stabilizing the 2,2′-azino-bis-3-ethylbenzothiozoline-6-sulfonic acid (IC₅₀ 1.23 mg/mL) and 1,1-diphenyl-2-picryl-hydrazil (DPPH) radicals (IC₅₀ 0.48 mg/mL) (p < 0.05). No significant differences in hydrogen peroxide (H₂O₂) scavenging and ferrous ion chelating properties of the EtOAc extracts of the seaweeds were apparent. The utilities of the reverse-phase high-performance liquid chromatography (RP-HPLC) method hyphenated to diode-array detection for analyzing the fingerprints of phenolic constituents in the solvent extracts and fractions of the seaweeds were denoted. High-performance liquid chromatographic analysis indicated the presence of phenolic acids in the solvent extracts of seaweeds. This study demonstrated the potential use of A. longifolius as candidate species to be used as a nutritional food supplement/functional foods to increase the shelf life of food items for human consumption.     

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。     

Biological Activities of Freshwater Algae,Spirogyra singularis Nordstedt

Spirogyra is commonly found as accessible algae in freshwater areas all over the world. Some medical uses have been reported from this genus. Biological activities of Spirogyra singularis were investigated employing eight in vitro assays. The extract showed different antioxidant activity. IC₅₀ for DPPH radical-scavenging was 4.71 ± 0.11 μg mL^?1. The extract showed very strong nitric oxide-scavenging activity with IC₅₀ = 77.3 ± 2.07 μg mL^?1, but its Fe²⁺ chelating ability was weak. The extract also exhibited high antioxidant activity in hemoglobin-induced linoleic acid peroxidation tests and scavenging of hydrogen peroxide. The extract also showed good antihemolytic activity. The total amount of phenolic content in the extract was determined as gallic acid equivalents, and total flavonoid content was calculated as quercetin equivalents. Biological activities may be attributed, at least in part, to the presence of phenol and flavonoid contents in the extract.     

Antioxidant Properties of Protein Hydrolysate Obtained from Oyster Saccostrea cucullata (Born, 1778)

The antioxidant activities of enzymatically hydrolyzed (protease from Bacillus cereus SU12) oyster (Saccostrea cucullata) protein were studied. The hydrolysate exhibited a strong antioxidant potential in 1, 1-diphenyl-2-picrylhydrazyl (DPPH, 85.7 ± 0.37%), followed by hydrogen peroxide radical scavenging activity (81.6 ± 0.3%), hydroxyl radical scavenging activity (79.32 ± 0.6%), and reducing power assay (2.63 ± 0.2 OD at 700 nm) at a concentration of 1 mg/mL. Due to the high antioxidant potential, the hydrolysate was purified in Sephadex G-25 gel filtration chromatography. The active peptide fraction was identified by DPPH and reducing power assay. The amino acid content of the purified active peptide fraction was analyzed by high performance liquid chromatography. The active fraction contained a good quantity of both essential and nonessential amino acids. The present study revealed that oyster (S. cucullata) protein hydrolysate is a potential source for natural antioxidants.     

Influence of Different Hydrolysis Processes by Trypsin on the Physicochemical,Antioxidant, and Functional Properties of Collagen Hydrolysates from Sphyrna lewini,Dasyatis akjei,and Raja porosa

ABSTRACT

Acid soluble collagen hydrolysates from the cartilages of Sphyrna lewini, Dasyatis akjei, and Raja porosa on different hydrolysis conditions by trypsin were prepared and named as S, D, and R, respectively. The hydrolysates (S1, D1, and R1) obtained from hydrolysis in pH 2.5, 3 h, 37°C presented wonderful foaming and emulsifying capacities, caused by their high average molecular weights (AMWs), high degree of Pro and Lys hydroxylation, and imino acid contents (17.1, 15.3, and 14.1%). At the concentration of 0.1% (w/v), the foaming capacities of S1, D1, and R1 were 104.75 ± 2.57, 63.87 ± 2.73, and 76.87 ± 2.02%; and the emulsifying activity indexes of them were 116.07 ± 1.89, 91.04 ± 1.79, and 123.85 ± 2.14 m²/g, respectively. Conversely, hydrolysates (S2, D2, and R2) obtained from hydrolysis in pH 7.8, 3 min, 37°C exhibited strong scavenging activity on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (EC₅₀ 0.568, 0.680, and 1.634 mg/mL), hydroxyl radical (EC₅₀ 0.253, 0.376, and 0.438 mg/mL), and moderate reducing power (0.0465–0.4702 at the concentration of 5 mg/mL), due to their low AMWs. The results indicated that S1, D1, and R1 could be used as emulsifiers and foaming agents, and S2, D2, and R2 could be used as natural antioxidants in food systems.     

Assessment of Bioactive Potential of Aqueous Protein Extracts from Diatoms Nitzschia laevis,Spirulina platensis,and Chlorella vulgaris

The bioactivities of protein extracts from Nitzschia laevis, Spirulina platensis, and Chlorella vulgaris were evaluated in vitro. Free radical scavenging potential, reducing power, oxygen radical absorbance capacity, superoxide anion radical, xanthine oxidase inhibition, and potential to inhibit angiotensin 1-converting enzyme (ACE) and acetylcholinesterase enzyme (AChE) were studied. Nitzschia protein extracts showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities, reducing power, and ACE inhibitory activity. Spirulina’s proteins showed the highest 2,2′-Azino-bis (3 ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, xanthine oxidase, and AChE inhibition activities. Aqueous protein extracts from Nitzschia, Spirulina, and Chlorella showed in vitro antioxidant, anti-ACE, and anti-AChE activities, suggesting possible new sources of bioactive proteins of different phyla with nutraceutical and pharmaceutical potentials.     

大眼金枪鱼头蛋白酶解物1 ku超滤组分的抗氧化活性及其理化性质

研究大眼金枪鱼头蛋白酶解物1 ku超滤组分体外的还原力、自由基清除能力及对衰老小鼠体内抗氧化能力的影响,分析1 ku超滤组分的一般成分、氨基酸组成及分子量分布,为进一步分离纯化金枪鱼头抗氧化肽提供基础。体外结果显示,1 ku超滤组分对羟基自由基、超氧阴离子和DPPH自由基的清除活性随浓度的增加而增强,IC50分别为1.38、0.73与0.93mg/mL,还原力也随浓度的增加而增大,在浓度为12.5 mg/mL时为0.763;体内结果显示,灌胃30 mg/kg的1 ku超滤组分连续42 d,D-半乳糖致衰老小鼠肝组织的超氧化物歧化酶(SOD)活性、肝组织和血清的谷胱甘肽过氧化物酶(GSH-PX)活性显著提高(P<0.05),血清丙二醛(MDA)含量显著降低(P<0.01);理化分析结果显示,1 ku超滤组分(干基计)蛋白质含量为96.40%,脂肪0.11%,灰分4.86%,疏水性氨基酸占氨基酸总量的35.8%,活性组分分子量在1 802~2 519 u和422~922 u。     

A Comparative Study on Some Functional and Antioxidant Properties of Kilka Meat,Fishmeal, and Stickwater Protein Hydrolysates

This study determined some antioxidative activities and functional properties of hydrolyzed proteins prepared from kilka (Clupeonella sp.) meat (KMH), kilka fishmeal (KFH), and stickwater (SWH). The highest degree of hydrolysis (DH) was 25.02% (p < 0.05) in SWH; however, the difference in DH between KMH and KFH was not significant. KFH had the highest protein content (85.69%) compared with other hydrolysis samples. The Fourier transform infrared spectra of KFH had the highest amide absorption picks (p < 0.05). Solubility at neutral pH was 84.09%, 86.00%, and 93.08% for KMH, KFH, and SWH, respectively. The highest fat absorption capacity was obtained with KFH (204 mg/50 mg). KFH also showed the best foaming properties with 25.66% foaming capacity and proper stability during 60 min (p < 0.05). The highest and lowest whiteness values were obtained in KMH (72.33%) and SWH (36.54%), respectively (p < 0.05). The highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) (IC₅₀ value of 1.99 mg/mL) and 2,2?-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) (IC₅₀ value of 2.00 mg/mL) radical scavenging activities were measured at the highest concentrations (40 mg/mL) in KFH (p < 0.05). KFH had the best functional and antioxidant properties, while SWH had a strong ability to inhibit lipid oxidation. Therefore, stickwater from fishmeal production factories is a potential resource for use in developing value-added products instead of being discarded.     

Optimized Extraction,Preliminary Characterization and Evaluation of the in Vitro Anticancer Activity of Phlorotannin-Rich Fraction from the Brown Seaweed,Cystoseira sedoides

Brown seaweeds produce useful bioactive substances with high cosmetic and pharmacological values due to the presence of antioxidant derivatives, mainly phlorotannins (PHT), which are of particular interest. A central composite rotatable design (CCRD) with three variables (extraction time, dry material-to-solvent ratio and ethanol concentration) and two responses was performed to optimize the microwave-assisted extraction (MAE) of phlorotannins from Cystoseira sedoides using response surface methodology (RSM). Results showed that the independent variables significantly affected both phlorotannin content and the scavenging capacity. The optimum operating conditions were extraction time, 101.74 sec; dry material-to-solvent ratio, 1:10 g/mL; and ethanol extraction, 50%. Under these conditions, the predicted values of PHT content and radical scavenging activity-IC50 were close to the observed values and were 383.887 µg PGE/g Dm and 18.353 µg/mL, respectively. Characterization of the phlorotannin-rich fraction was conducted by high-performance liquid chromatography and Fourier transform infrared spectroscopy. The evaluation of the anticancer activity against MCF-7 breast cancer cell line showed a potent activity to trigger apoptotic death in more than a half of the MCF-7 cells, with an estimated IC50 value of 78 μg/mL. In addition, this fraction induced a notable growth regression effect on 3D spheroids model in a concentration-dependent manner, with a growth rate of about 1.17, at 200 µg/mL.

Abbreviations: CCRD: Central composite rotatable design; Dm: Dry material; DMBA: 2,4-dimethoxy benzaldehyde; DMSO: Dimethyl sulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EDTA: Ethylene diamine tetraacetic acid; FBS: Fetal bovine serum; FT-IR: Fourier transform infrared; HPLC: High-performance liquid chromatography; IC₅₀: Half-maximal inhibitory concentration; MAE: Microwave-assisted extraction; Min: minutes; PBS: Phosphate buffered saline; PGE: Phloroglucinol equivalent; PHT: Phlorotannins; PHT-SED: phlorotannins derived from Cystoseira sedoides; PI: Propidium iodide; RSA: Radical scavenging activity; RSM: Response surface methodology; Sec: seconds     

Assessment of Aquatic Aeromonas spp. Isolates' Susceptibility to Cinnamaldehyde,Vanillin, and Crude Kigelia africana Fruit Extracts

Aeromonas spp. are primary and opportunistic fish pathogens that are responsible for severe economic losses in aquaculture, and are exacerbated by their increasing incidence of antimicrobial resistance. Phytochemicals are being explored as alternatives to conventional antimicrobial agents since they have destressing, growth‐promoting, immune‐stimulating, and antimicrobial properties. The susceptibility of 93 aquatic Aeromonas spp. and six Plesiomonas shigelloides isolates (from cultured catfish, koi carp, tilapia, and seawater) to three phytochemicals, viz.: cinnamaldehyde (10–1250 µg/mL), vanillin (5–250 µg/mL), and crude Kigelia africana fruit extracts (4–10 mg/mL ethyl acetate, dichloromethane, methanol, and hexane) were assessed using the disk diffusion assay and compared to ampicillin and tetracycline, using activity indices. No zones of inhibition were obtained with 10 µg/mL of cinnamaldehyde, or with 5–250 µg/mL of vanillin. However, varying degrees of inhibition were observed with higher concentrations of cinnamaldehyde, as well as with the K. africana methanol extract. Cinnamaldehyde (≥500 µg/mL) and the K. africana methanol extract displayed better antimicrobial activity against study isolates in comparison to vanillin and ampicillin. They appear to be promising and sustainable phytochemicals that might be useful as alternatives to the antimicrobial agents currently in use in aquaculture.     

5种中草药的不同溶剂提取物对嗜水气单胞菌抑菌活性的研究

为探讨不同有机溶剂对常用中草药的提取效果及其提取物对水产常见病原菌的抑菌活性，以50%乙醇、无水乙醇、氯仿、甲醇、丙酮、乙酸乙酯和石油醚为提取溶剂，根据水浴旋转蒸发器回流提取工作原理，分别对五倍子、乌梅、诃子、大黄和五味子等5种中草药进行固液萃取，并用其提取物对水产高致病性病原菌嗜水气单胞菌（Aeromonas hydrophila）进行体外抑杀菌试验。结果表明：50%乙醇、无水乙醇、甲醇对5种中草药的提取效果优于其他4种溶剂，尤以甲醇的提取效果最佳，其提取率分别为五倍子46.09%、乌梅27.58%、诃子29.55%、大黄50.34%、五味子30.88%;提取效果最差的是石油醚，其提取率分别为五倍子1.98%、乌梅1.73%、诃子2.21%、大黄0.89%、五味子3.40%。体外抑菌试验结果显示，嗜水气单胞菌对五倍子的3种醇提物均表现为极度敏感，对大黄的甲醇提取物表现为高度敏感，对五倍子、大黄和五味子的氯仿提取物以及对乌梅、大黄和五味子的石油醚提取物呈低度敏感，对五倍子的丙酮提取物等共25种提取物表现为中度敏感；五倍子、大黄的醇提物对供试菌的抑杀菌效果明显优于其他3种中草药的任何一种...     

Hypnea musciformis: alternative or complement to the production of Kappaphycus alvarezii introduced in tropical countries?

Hypnea musciformis that are harvested from natural beds and introduced Kappaphycus alvarezii are the two main sources of raw material for the production of carrageenan in Brazil. The daily growth rate (DGR) and carrageenan yield (CY) of both species were evaluated in vitro and in the sea to assess the feasibility of cultivating H. musciformis as a complement or as an alternative to the production of K. alvarezii. The DGR of H. musciformis (10.8 ± 0.6% day^?1) was higher than the DGR of K. alvarezii (5.6 ± 0.3% day^?1) in vitro, and the CY extracted using NaOH (62.3 ± 0.15%) was better than using KOH (32.0 ± 0.3%). In eutrophic medium, the DGR of H. musciformis (9.4 ± 2.1% day^?1) was higher than that of K. alvarezii (5.0 ± 1.2% day^?1). However, in this condition, the CY was similar for both species (50.2 ± 3.5%), and both species were effective in removing nutrients (30% NH₄⁺, 18% NO^2? and 2.4% ). H. musciformis did not survive when cultivated in the sea, neither in monospecific nor in multispecific cultivation. Moreover, K. alvarezii grew better in monospecific cultivation throughout the year (from 1.73 to 5.15% day^?1) except during the wet period and the CY was high in both cultivation types (33.7–50.8%). We conclude that the cultivation of H. musciformis is not an alternative to K. alvarezii on the Brazilian southeastern coast. However, the results in vitro suggested its use as a complementary source of carrageenan or as a biological filter.     

Utilization of Chum Salmon (Oncorhynchus keta) Skin Gelatin Hydrolysates to Attenuate Hydrogen Peroxide-Induced Oxidative Injury in Rat Hepatocyte BRL Cell Model

Six chum salmon skin gelatin hydrolysates with degree of hydrolysis of 4.7–13.5% were generated by Alcalase and papain and showed scavenging activities to 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and superoxide anion radicals. Potential protection of these hydrolysates against H₂O₂-induced oxidative injury in rat hepatocyte line (BRL cells) was assessed in vitro, based on cell viability, leakage of lactate dehydrogenase, thiobarbituric acid reactive substances (TBARS), glutathione content, glutathione reductase, and catalase of the cells. The hydrolysates exhibited protection on the hepatocytes, reflected by an enhanced cell viability (about 6.2–87.9%), glutathione reductase (about 28.2–85.9%) or catalase (about 43.3–116.9%) activity, and decreased lactate dehydrogenase leakage (about 2.7–34.7%) or TBARS content (about 4.1–39.3%). On the contrary, cellular glutathione content gave an unnoticeable difference among the investigated groups. These hydrolysates also showed protection against DNA damage in the cells. Cytoprotection of these hydrolysates possessed a dosage-dependence manner. More importantly, there was a high correlation (0.7 < r < 0.9) between DPPH or superoxide anion radical scavenging activity of these hydrolysates and three cellular antioxidant indices (glutathione content, glutathione reductase, and catalase activity).     

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.     

Antioxidant Potential of Water Hyacinth (Eichornia crassipes): In Vitro Antioxidant Activity and Phenolic Composition

The aims of the present study were (a) to extract and quantify the main phenolic acids and tocopherols from the petiole, leaves, and flowers of Eichornia crassipes; (b) to evaluate the antioxidant capacity of the extracts in four in vitro systems (1,1-diphenyl-2-pycryl-hydrazyl [DPPH] radical scavenging ability, iron chelating activity, reducing power, and prevention of oxidation in a liposome model system); and (c) its effectiveness in retarding lipid peroxidation in fish oil by accelerated stability test. Significant differences were observed in total and individual phenolic contents and in the antioxidant activities of extracts from the various parts of E. crassipes. Out of the 11 phenolic acids analyzed, ethanolic extracts contained high amounts of gallic, protocatechuic, gentisic, and p-hydroxybenzoic acid, whereas, water extracts contained less amounts of a varied number of phenolic acids. Ethanolic extracts of flower, which contained the highest total phenolic content, were found to have high DPPH radical scavenging activity and reducing power. However, ethanolic extracts of leaf exerted a high Fe²⁺ chelating activity and also inhibited lipid peroxidation process both in liposomes and fish oil. Our results demonstrate that E. crassipes, an underutilized aquatic weed, could be a potential natural antioxidant source for food, feed, and pharmaceutical applications.     

浒苔提取物对太平洋牡蛎受精卵孵化的抑制效应

针对2007-2009年间在黄海海域形成"绿潮".的大型绿藻浒苔(Enteromorpha prolifera).研究了其3种不同极性有机溶剂(甲醇、丙酮、氯仿)提取物对太平洋牡蛎(Crassostrea gigas)受精卵孵化的影响.结果表明.在48 h内.浒苔的甲醇和丙酮提取物对太平洋牡蛎受精卵孵化有较强的抑制作用.高浓度提取物能够完全抑制太平洋牡蛎受精卵孵化至D型幼虫.而氯仿提取物对太平洋牡蛎受精卵孵化的影响没有明显的浓度-效应关系.应用乙酸乙醋和石油醚对浒苔的甲醇提取物进行液-液萃取.并探讨了乙酸乙醋相和石油醚相中的浒苔提取物对牡蛎受精卵孵化的影响.结果表明.石油醚相和乙酸乙醋相中的浒苔提取物对牡蛎受精卵孵化均有很强的抑制效应.上述研究表明.浒苔中存在能够抑制太平洋牡tfi受精卵孵化的活性物质.这类物质应具有较高的极性.     

Functional Properties of the Marine Gastropod Molluscs Chicoreus ramosus and Babylonia spirata Collected from the Southern Coast of India

ABSTRACT

This study characterized the functional properties of ethyl acetate-methanol (EA-MeOH) and chloroform extracts of the muricid gastropod Chicoreus ramosus and buccinid Babylonia spirata. The EA-MeOH extract of B. spirata was a potent quencher of radical cation (IC₉₀ < 1 mg/mL), whereas that of C. ramosus held greater anti–inflammatory potential (IC₉₀ 5-lipoxygenase inhibition 1.74 mg/mL) than the buccinid gastropod. The EA-MeOH extract of C. ramosus displayed potent antidiabetic activities as deduced by its attenuation properties against carbolytic enzymes α-amylase and α-glycosidase (IC₉₀ 1.06 and 2.25 mg/mL, respectively) than those exhibited by B. spirata (IC₉₀ 2.32 and 4.50 mg/mL, respectively). The spectroscopic dereplication studies to determine probable chemical groups in the solvent extracts of the studied gastropods revealed the presence of functionalities, which might augment the electronic properties of the bioactive principles present within the extract, thereby enhancing their activity. Thus, the present study recognized that marine gastropods C. ramosus and B. spirata have potential functional implications against oxidative stress-induced diseases, such as diabetes and inflammation. This research supports further investigation of these previously uncharacterized marine gastropod species to isolate potential bioactive leads for use against various biological targets and to develop functional food formulations.     

胰蛋白酶酶解制备孔鳐软骨蛋白寡肽及其抗氧化活性

以孔鳐软骨为材料,采用盐酸胍抽提、丙酮分级沉淀,制备孔鳐软骨蛋白;以DPPH·和HO·清除活性为导向,采用胰蛋白酶酶解、膜超滤、DEAE-52阴离子交换层析、Sephadex G-15凝胶层析和反相高效液相色谱(RP-HPLC)等技术,制备抗氧化肽,并对其活性进行系统评价。结果显示,孔鳐软骨蛋白经胰蛋白酶酶解和分离纯化得到2个抗氧化肽RCPE-A和RCPE-B,经氨基酸序列分析确定其序列分别为Gly-Glu-Glu-Gly-Pro-Arg-Gly (GEEGPRG)和Gly-Glu-Glu-Gly-Thr-Met-Gly-Leu (GEEGTMGL),质谱(ESI-MS)测定其分子量分别为700.71和792.87 u。体外自由基清除实验结果显示,RCPE-A与RCPE-B对DPPH·(EC₅₀ 2.94和1.16 mg/mL)、HO·(EC₅₀ 0.34和0.54 mg/mL)、ABTS⁺·(EC₅₀ 0.34和0.10 mg/mL)和O₂^-·(EC₅₀ 0.11和0.03 mg/mL)具有良好的清除作用,RCPE-A与RCPE-B亦显示出较强的脂质过氧化抑制作用。研究表明,孔鳐软骨蛋白酶解物及制备多肽可用于抗氧化相关的功能食品开发,也可以用作抗氧化剂延长相关产品的货架期。     

Utilization of Shrimp By-Products by Bioconversion with Medical Fungi for Angiotensin I-Converting Enzyme Inhibitor and Antioxidant

ABSTRACT

A liquid fermentation process to bioconvert shrimp by-products was developed with two species of fungi, Boletus edulis and Suillus bovinus. Angiotensin I-converting enzyme (ACE) inhibitory, antioxidant activities of bioconverted products and the deodorization effect of fungal bioconversion were determined. Two species of fungi could grow quickly in shrimp by-products medium and deodorize shrimp by-products. Bioconverted products had higher ACE inhibitory and antioxidant activities than control medium. Water extracts without crude polysaccharides from mycelia of B. edulis had the highest ACE inhibitory activity (IC₅₀ = 0.084 ± 0.011 mg/mL). Crude exopolysaccharides of B. edulis had higher antioxidant activity. Bioconversion with the fungus was proven to be a novel, practicable way to recycle shrimp by-products.