胶州湾菲律宾蛤仔的污染现状和大肠菌群净化研究

2003年 4~10月间，对山东省胶州湾内 6个采样点分 4次采集的菲律宾蛤仔 (Ruditapes philippinarum)进行了 Pb、As、Cu、Cd、Hg及大肠菌群的调查分析。结果显示，重金属离子的含量基本符合标准，但大肠菌群普遍超标。针对这一问题，提出应对超标的菲律宾蛤仔采取先净化后上市的措施，以提高菲律宾蛤仔的食用安全。     

培育密度对菲律宾蛤仔浮游幼虫生长与成活的影响

试验结果表明:菲律宾蛤仔浮游幼虫试验密度为5,10,15,20个/ml,各组幼体的平均日生长速度依次为18 7,12 1,11 2,8 3μm,成活率依次为88 0%,82 0%,79 8%,50 4%。综合分析认为,菲律宾蛤仔浮游幼虫在生产中育苗密度以10个/ml,15个/ml较为适宜。     

In Vivo Antioxidant Effects of Hydrolysate Derived from Waste Proteins of Mactra veneriformis

Mactra veneriformis has been used as seafood and traditional Chinese medicine for thousands of years in China. In the present study, hydrolysate was prepared from waste proteins of M. veneriformis by trypsin treatment. The protective effect of hydrolysate on the defense system against oxidative stress was investigated by estimating the superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in normal mice. The antioxidant activity of hydrolysate was investigated in D-galactose-induced aging mice by estimating the SOD and glutathione peroxidases (GSH-px) activity and MDA level. Amino acid composition and molecular weight distribution were also determined. These investigations suggest that hydrolysate derived from by-products of M. veneriformis possess antioxidant activity and may be attributed to elevation of the activity of antioxidase and enhanced antioxidant defenses.     

Improvement of Antioxidant Activity of Grass Carp (Ctenopharyngodon idella) Protein Hydrolysate by Washing and Membrane Removal Pretreatments and Ultrasonic Treatment

To improve the antioxidant activity of grass carp hydrolysate, washing and membrane removal pretreatments and ultrasonic treatment were applied in this study. Various pretreatment methods (washing methods and membrane removal methods) for removing the prooxidants and phospholipid in minced fish were evaluated. The effects of ultrasonic treatment prior to enzyme hydrolysis on the antioxidant activities of minced carp hydrolysate were investigated. The minced carp subjected to washing with 0.4% NaCl solution and subsequent membrane removal with CaCl₂ and citric acid solution had the lowest content of prooxidants and phospholipid remaining in the minced fish. The hydrolysate produced from washing/membrane removal pretreated carp with ultrasonic treatment for 20 min had the highest ferric reducing antioxidant power and scavenging activities against 1,1-diphenyl-2-picrylhydrazyl and 2,2?-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radicals. This suggested that the combination approach of washing/membrane removal pretreatment and ultrasonic treatment could improve the antioxidant activity of carp hydrolysate, and this hydrolysate could be a potential antioxidant ingredient for functional foods.     

Purification and Identification of Antioxidant Peptides from Fermented Fish Sauce (Budu)

This study aimed to isolate antioxidant peptides from Budu extract and identify novel antioxidant peptides using high-performance liquid chromatography (HPLC) and electrospray ionization-time-of-flight mass spectrometry (ESI-TOF MS/MS). An amino acid analyzer was used to analyze the amino acid composition of peptides. Two novel peptides, Lue-Asp-Asp-Pro-Val-Phe-Ile-His (LDDPVFIH) and Val-Ala-Ala-Gly-Arg-Thr-Asp-Ala-Gly-Val-His (VAAGRTDAGVH), were identified. The synthesized peptide of LDDPVFIH showed higher antioxidant activity. The presence of hydrophobic amino acids (Ile and Leu), acidic (Asp) and basic (His) amino acids in the peptide sequences is believed to contribute to the high antioxidant activity of the fermented anchovy fish (Budu) extract. Thus, the two peptides may have potential application as functional foods and could also be used as nutraceutical compounds.     

Antioxidant Assessment of Schizochytrium Meal Protein Enzymatic Hydrolysate and Its Potential Application

Schizochytrium meal protein (SMP) extracted from Schizochytrium meal was hydrolyzed by flavourzyme. Response surface methodology (RSM) was used to optimize the extraction conditions for the protein extraction yield from Schizochytrium meal. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (DRSA) was used to evaluate the antioxidant activity of hydrolysates. The orthogonal test was used to investigate the effects of three independent variables, namely protease dosage, hydrolysis time, and pH. The optimum conditions obtained were as follows: protease dosage of 7%, hydrolysis time of 1.5 h, pH of 6, under which, DRSA at the concentration of 2 mg/mL was 89.38%. Aspartic and glutamic acid constituted approximately 26.32% of the total amino acids, and glutamic acid was the most abundant amino acid of Schizochytrium meal protein hydrolysate (SMPH) by amino acid composition analysis, which may have contributed to the scavenging activity of SMPH. Moreover, SMPH was made into chewable tablets with suitable formula and high humidity stability. These findings indicate that Schizochytrium meal can be reused as a raw material for preparation of antioxidant peptides.     

Purification and Characterization of a New DPPH Radical Scavenging Peptide from Shrimp Processing By-products Hydrolysate

A peptide with high antioxidant activity was isolated and identified from shrimp processing by-products hydrolysate. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity was used to evaluate the antioxidant activity of fractions. The purified antioxidant peptide was identified as Ser-Val-Ala-Met-Leu-Phe-His (804.4 Da) by electrospray ionization tandem mass spectrometry. The purified peptide at 50 μg/mL showed antioxidative activity of 65.7 ± 0.9%, which was 3.18-fold higher compared with the first step separation by ion-exchange chromatography. It is possible to produce natural antioxidative peptides from shrimp processing by-products hydrolysate. The high antioxidant activity may be due to the presence of Phe-His segment at the C-terminus of the peptide.     

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.     

Antioxidative Activitiy of Protein Hydrolysate from the Muscle of Common Kilka (Clupeonella cultriventris caspia) Prepared Using the Purified Trypsin from Common Kilka Intestine

Trypsin from the intestine of common kilka (Clupeonella cultriventris caspia) was purified using ammonium sulfate precipitation (30–50% saturation), Sephadex G-75, and DEAE-cellulose chromatography with the purity of 30-fold and the yield of 12%. The molecular weight of trypsin was estimated to be 23.2 kDa based on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The trypsin had optimal activity at pH 8.0 and 60°C using N-α-benzoyl-DL-arginine-ρ-nitroanilide hydrochloride (BAPNA) as a substrate and showed high stability in the pH range of 7.0–10.0. It was stable up to 50°C. Soybean trypsin inhibitor (SBTI) and N-ρ-tosyl-L-lysine-chloromethylketone (TLCK) significantly inhibited trypsin activity (p < 0.05). Protein hydrolysate from common kilka muscle with different degrees of hydrolysis (DHs; 20, 30, and 40%) was prepared using the purified trypsin, and antioxidative activities were determined. 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing antioxidant power, and ferrous chelating activity of hydrolysate increased with increasing DH up to 40% (p < 0.05). Therefore, trypsin from intestine of common kilka could be used as a processing aid for production of fish protein hydrolysate with antioxidative activity.     

Purification and Identification of an Antioxidative Peptide from Digestive Enzyme Hydrolysis of Cutlassfish Muscle

ABSTRACT

The major objective of this study was to investigate the pepsin digests of cutlassfish muscle for their antioxidant activity. The peptide isolated after 3 h of pepsin hydrolysis demonstrated the highest scavenging activity for both 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and alkyl radicals. It also had the highest yield of the isolates with the highest protein content, which was likely related to its higher degree of hydrolysis than the other hydrolysates. The antioxidant peptide purified from the 3 h pepsin hydrolysate indicated the amino acid sequence, Phe-Ser-Gly-Gly-Glu. This study suggests that cutlassfish muscle is an excellent antioxidant source and could be effectively used as a food ingredient and in pharmaceuticals.     

Antioxidant and ACE Inhibitory Activities of Kawakawa (Euthynnus affinis) Protein Hydrolysate Produced by Skipjack Tuna Pepsin

ABSTRACT

Pepsin enzyme from skipjack tuna was extracted for the production of kawakawa (Euthynnus affinis) fish protein hydrolysate. Using ultra-fractionation, Kawakawa protein hydrolysates were separated into four different fractions, including fractioned protein hydrolysate I (FPH I) (< 1 kDa), FPH-II (1–3 kDa), FPH-III (3–10 kDa), and FPH-IV (> 10 kDa). The antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, inhibition of linoleic acid oxidation, reducing power tests, and chelating activity of metal ions. Results indicated that FPH II fraction peptides had higher antioxidant activity in comparison with the other fractions, followed by FPH I. Further, the fractions were evaluated for angiotensin converting enzyme (ACE) inhibition, and IC₅₀ value ranged from 0.45 to 1.86 mg/ml with higher activity in FPH I (IC₅₀ 0.45). Finally, the amino acid profile of different fractions was analyzed. The fractions exhibited significant amounts of hydrophobic amino acids, which could perform as hydrogen donors, frustrate the free radicals, and inhibit the ACE. The recovered pepsin from the viscera was used to produce hydrolysates with good biological activities. Peptides lower than 3 KDa had antioxidant activity as positive controls and significant ACE activity. These are very important findings that could be used to conduct further research in a preclinical study of these peptides.     

In Vitro Comparative Evaluation of Antioxidant Activity of Hydrophobic and Hydrophilic Extracts from Algicolous Fungi

The screening of potential antioxidant activities of hydrophobic (ethyl acetate) and hydrophilic (polysaccharide) extracts from both mycelia and fermentation media of eleven algicolous fungi was performed using three antioxidant assays. Algicolous fungi were able to ferment low-cost nutrients composed of potato infusion, glucose, and natural seawater and produce hydrophobic and hydrophilic metabolites with antioxidant properties. Signi?cant differences were observed in antioxidant activities of extracts from the various species evaluated using total antioxidant activity (TAA), hydroxyl radical scavenging activity (HRSA), and ferric reducing antioxidant power (FRAP), as well as their total sugar contents (TSC) of hydrophilic extracts and total phenolic contents (TPC) of hydrophobic extracts. TSC of endopolysaccharides was higher than exopolysaccharides and showed a significant correlation with both TAA and FRAP. Most of mycelial extracts were richer in phenolics than fermentation media, and TPC of hydrophobic extracts was significantly correlated with their FRAP. Mycelial extracts for most of algicolous fungi showed higher antioxidant activities than fermentation media. Both hydrophobic and hydrophilic extracts of algicolous fungi showed good antioxidant properties, especially as hydrogen-donating antioxidants, which could be considered for future applications in medicine, food production, or cosmetic industry.     

Taste Compounds and Biofunctional Activities of the Sandy Beach Clam Hydrolysate for the Shellfish Flavoring Condiment

ABSTRACT

Sandy beach clam meat was hydrolyzed with proteases (Alcalase and Flavozyme) for the manufacture of a natural flavoring condiment. The optimal conditions for the enzymatic hydrolysis of the shellfish meat via the response surface methodology were: temperature, 54.7°C; pH, 5.9; time, 45 h. The total amino acid content of the shellfish hydrolysate was 9,218.8 mg/100 g and consisted mainly of glutamic acid, aspartic acid, glycine, arginine, lysine, and leucine. Nucleotides and their related compounds of the shellfish hydrolysate consisted mainly of adenosine diphosphate (0.86 mg/100 g), adenosine monophosphate (11.24 mg/100 g), and inosine (49.03 mg/100 g). The shellfish hydrolysate resulted in three protein peaks in the Bio-Rad P2 gel chromatography pattern. The antioxidant activity was highest at Peak 1 (64.67%), followed by Peak 2 (27.48%) and Peak 3 (16.67%); whereas the angiotensin I-converting enzyme inhibitory activity was highest at Peak 3 (67.27%), followed by Peak 2 (6.14%). Antitumor activity was highest at Peak 1 (18.77%), followed by Peak 2 (8.75%). All the sensory attributes of the shellfish condiment were scored highly. Therefore, the sand beach clam hydrolysate can be commercialized as an ingredient of a natural flavoring condiment or seasoning.     

缢蛏多糖的提取及抗氧化作用研究

以新鲜缢蛏的碎肉为材料,加水配制成一定比例的匀浆,利用超声波提取法对缢蛏碎肉匀浆进行多糖的提取。利用蒽酮—硫酸法对多糖得率进行计算。将得到的多糖粗品通过DEAE离子交换层析及G-75凝胶层析进一步分离纯化。通过邻苯三酚自氧化体系和Fenton体系测定所得样品的抗氧化能力。结果表明:利用超声波提取多糖的提取率为19.93%。多糖粗品通过DEAE离子交换柱,得到2个峰,峰1对超氧阴离子的清除能率为43.89%,质量浓度为1 mg/ml时,对.OH清除率为57.80%。峰2对超氧阴离子的清除能率为57.80%,质量浓度为1 mg/ml时,对.OH清除率为77.83%。将DEAE峰2进行G-75凝胶层析得到2个峰,峰1对超氧阴离子的清除能率为83.08%,质量浓度为1 mg/ml时,对.OH清除率为94.76%。峰2对超氧阴离子的清除能率为74.41%,质量浓度为1 mg/ml时,对.OH清除率为52.41%。     

Effect of Enzyme Type on the Antioxidant Activities and Functional Properties of Enzymatic Hydrolysates from Sea Cucumber (Cucumaria frondosa) Viscera

ABSTRACT

Effects of various proteases were obvious on the amino acid composition, antioxidant activities, and functional properties of enzymatic hydrolysates prepared from sea cucumber viscera (SCV). Alcalase, trypsin, and flavourzyme conferred better antioxidant activities on hydrolysates and endowed them with a higher degree of hydrolysis, coming to 19.08, 32.38, and 15.94%, respectively. For functional properties, neutrase- and flavourzyme-derived hydrolysates showed better foaming properties. Amino acid composition suggested that papain, neutrase, alcalase, and bromelain granted the hydrolysate’s high nutritional quality. The results indicated that SCV hydrolysates could be used in food systems to impart desirable characteristics to products.     

辽宁菲律宾蛤仔土池育苗存在的问题与对策

菲律宾蛤仔Ruditapesphilippinarum(AdamsetReeve)也称蛤仔,辽宁部分地区俗称沙蚬子,是我省最主要的滩涂贝类养殖品种。近年来,为满足苗种供应严重不足的问题,沿海各产区相继开展了土池育苗生产实验工作、总体看,效果不理想。笔者总结近年来从事蛤仔人工育苗及土池育苗的经     

Purification and Characterization of an Antioxidant Protein from Pearl Oyster (Pinctada fucata martensii)

Protein from pearl oyster (Pinctada fucata martensii) was purified and characterized. Results showed that the pearl muscle protein (PMP) was composed of a single polypeptide with a molecular weight of approximate 11.6 kDa and contained 95.1% of the protein and 4.92% of the carbohydrate of the oyster. The polypeptide appears to be a glycoprotein, since the Fourier transform infrared (FTIR) spectroscopy of the PMP shows the typical characteristics of protein and polysaccharides. The denaturation temperature of the PMP was 81.3°C by differential scanning calorimetry. Circular dichroism spectroscopy showed that the PMP has a highly ordered structure. The PMP (200 μg/mL) showed a high oxygen radical absorbance activity (ORAC) of 6.57 μmol Trolox/μmol protein. Its antioxidant activity was stable at temperatures from 30–80°C and pH 2–10. The antioxidant activity was significantly inhibited by Zn²⁺, Ca²⁺, and EDTA and enhanced by Mn²⁺. The results suggest that the PMP could be used as a potential natural antioxidant.     

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.     

Growth,mortality and reproduction of the transplanted Manila clam (Ruditapes philippinarum Adams & Reeve 1850) in Jiaozhou Bay

Samples of Manila clam (Ruditapes philippinarum Adams & Reeve 1850) were collected from May 2004 to April 2005 monthly, and plankton net trawling of planktonic larvae and bottom sediment sampling surveys were further conducted from May to October 2006 in Jiaozhou Bay. Based on the data collected, growth, mortality and reproduction of the transplanted Manila clam and the environmental effects were examined. The results showed that the enhanced clams grew well and showed a growth trend similar to the local wild ones. The main growth periods lasted from April to September, with the water temperature being the main factor affecting the growth, which was the same as that of the wild clams. There were also two reproduction cycles for the farmed Manila clams each year in Jiaozhou Bay and the main breeding period was from May to June. The phenomenon of delayed metamorphosis was quite common through larval development. The farmed clams could spawn when they reached sexual maturity, but they could not perform effective recruitment as many planktonic larvae died during metamorphosis and settlement. A preliminary study indicated that sediment perturbation and marine environment pollution were the main factors causing the death of larvae in the development process.     

Evaluation of the Antioxidant Activity In Vitro and in Hippocampal HT-22 Cells System of Protein Hydrolysates of Common Carp (Cyprinus carpio) By-Product

Fish processing by-products may become more than 50% of the starting material. If mismanaged, these large quantities of discarded fish can create serious pollution problems and can also generate cost associated with their disposal. Enzymatic hydrolysis is one of the techniques that is currently being developed in order to recover and add value to these biomolecules. There is an increasing interest in natural antioxidants that are safer for consumers compared with synthetic antioxidants. In this study, common carp by-product was hydrolyzed using the enzymes Alcalase (A) and Protamex (P) to reach degrees of hydrolysis (DH) of 10 and 15%, respectively. Antioxidant capacity against peroxyl radicals (ACAP); 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging; and the measurement of intracellular reactive oxygen species (ROS) concentration after exposure to common carp protein hydrolysates were investigated. The results revealed that the hydrolysate A15 exhibited significantly (p < 0.05) higher antioxidant activity against the DPPH radical. A15 showed the highest in vitro antioxidant competence against peroxyl radicals, whereas P15 showed the lowest activity against peroxyl radicals (p < 0.05). Hydrolysates having the highest and the lowest in vitro antioxidant activity (A15 and P15, respectively) were selected for the determination of antioxidant activity in the HT-22 cells system. Measurement of intracellular ROS concentration revealed that P15 at the concentration of 1.25 mg/mL significantly (p < 0.05) reduced the intracellular ROS concentration. These results showed that common carp by-product protein hydrolysates are a source of antioxidant peptides with a high potential for food and pharmaceutical industries to develop new nutraceuticals and functional foods.