蛹虫草固体培养残基中SOD的提取研究

以酶总活力为指标对蛹虫草培养残基中的超氧化物歧化酶(SOD)提取条件进行研究,通过对培养残基与磷酸缓冲液的用量比即料液比、磷酸缓冲液pH值、提取温度、固体(NH4)2SO4饱和度这4个因素进行单因素试验与L9(34)多因素正交试验,发现料液比对SOD的提取效果影响最明显,其次为固体(NH4)2SO4饱和度和提取温度,而缓冲液pH值的影响相对较小.确定蛹虫草培养残基中SOD的最佳提取条件:料液比为1:20,提取温度为50℃,缓冲液pH值为7.6,固体(NH4)2SO4饱和度为40%,在这样的提取条件下测出的酶总活力最高,提取效果最好.     

超声辅助提取水飞蓟粕蛋白

为提高水飞蓟粕的附加值,采用超声波辅助提取水飞蓟粕中的蛋白质;采用单因素试验考察了液料比、磷酸盐缓冲液pH值、超声时间与间歇时间之比、超声波功率、提取时间及粒度等因素对超声提取水飞蓟粕蛋白的影响,确定了较优提取条件为:液料比为12、超声时间与间歇时间之比为3、缓冲液pH值8、提取时间60min、超声波功率800W,温度45℃,水飞蓟粕粒度20目.在此条件下,水飞蓟粕蛋白的表观提取率为49.1%,实际提取率为34.37%.     

巴西松子中蛋白酶的分离纯化及酶学性质

在单因素试验基础上,通过正交试验研究pH值、提取时间和料液比3个因素对巴西松子中蛋白酶活力的影响,得出巴西松子中蛋白酶的最佳提取缓冲液为pH9.0的硼酸-硼砂缓冲液、提取时间为60min、料液比为1:8(m/V);采用(NH4)2SO4沉淀、DEAE-Sepharose FF阴离子交换层析分离纯化巴西松子中的一种蛋白酶,结果表明:纯化后蛋白酶比活力提高到了8.61倍,回收率为21.65%。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析表明:该蛋白酶分子质量为33kD。酶学性质结果表明:该蛋白酶最适pH值为9.0,属碱性蛋白酶;反应的最适温度为50℃;金属离子Mn2+对该蛋白酶活性有强烈的激活作用,而Ca2+、Mg2+和Cu2+对酶活性有抑制作用。     

微波法提取鹿角漆树果穗红色素的工艺研究

采用微波法提取鹿角漆树果穗红色素,采用单因素试验法和正交试验法考察了提取剂的类型、pH值、料液比、微波功率、提取时间5个因素对鹿角漆树果穗红色素提取的影响。试验结果表明:最佳提取溶剂为50%乙醇。微波提取鹿角漆树果穗红色素的各因素影响顺序依次为:微波功率pH值提取时间料液比;优化工艺条件为提取时间5min,微波功率为320W,pH值为5,料液比为1:30。     

响应面优化可溶态和膜结合态马铃薯多酚氧化酶提取工艺

以大西洋马铃薯为原料,分别用柠檬酸-磷酸盐缓冲液浸提和超声波辅助Tris-HCl缓冲液浸提法提取马铃薯中可溶态(sPPO)和膜结合态(mPPO)多酚氧化酶,以酶比活力为评价指标,在单因素实验基础上,通过响应面对sPPO和mPPO的提取工艺进行优化。结果显示:sPPO最优提取工艺为料液比1:3.2、浸提时间12 h、缓冲液pH7.1,在该条件下测定sPPO比活力平均值为(130.5±2.4) U/mg,三个因素对sPPO提取的影响大小依次为:浸提时间>料液比>缓冲液pH。mPPO最优提取工艺为料液比1:5.2、浸提时间7 h、缓冲液pH6.7,在该条件下测定mPPO比活力平均值为(686.4±7.9) U/mg,三个因素对mPPO提取的影响大小依次为:超声时间>料液比>缓冲液pH。所得响应面模型可以很好地预测和分析sPPO和mPPO提取工艺条件。     

响应面法优化超声波辅助复合酶法水解虾头蛋白的工艺

为充分提取虾头蛋白以提高虾头的利用价值,本研究以超声波辅助蛋白酶水解南美白对虾虾头蛋白,探究了蛋白酶类型、蛋白酶添加量、料液比、pH值、超声功率、超声时间、酶解温度和酶解时间等8个因素对虾头蛋白水解度的影响,并通过响应面法优化获得最佳的超声和酶解条件。结果得出,最佳提取工艺条件为:虾头泥以料液比1∶10(g/mL)调配→pH调节为7.5→超声功率180 W下处理30 min→料液中添加1%碱性蛋白酶和0.1%中性蛋白酶→50℃下酶解时间3 h→加热煮沸10 min→4000 r/min下离心20 min→取上清液,即水解蛋白液;该工艺条件下,虾头蛋白水解度可达到42.5%。     

从米糠中提取植酸钙的工艺研究

对米糠中提取植酸钙的工艺进行了研究.以酸浸pH值、浸泡时间、液料比为影响因素,进行了3因素3水平正交试验.经多次重复试验,结果表明最佳工艺为:酸浸pH值2.0,浸泡时间5h,液料比8∶1;在最佳条件下,植酸钙的提取率为9.06％.通过对植酸钙标准品和所提取物质的红外谱图的比较,证实所提取物质为植酸钙粗品.     

决明子蛋白质提取工艺研究及其体外消化率的测定

目的:优选决明子蛋白质的最佳提取工艺及体外消化率测定.方法:采用正交试验法,以凯氏定氮法测得决明子蛋白质的含量和得率为考察指标,对提取过程中缓冲液浓度、pH值、固液比及提取时间4个工艺因素进行优化研究.并用胃蛋白酶-胰蛋白酶法测定体外消化率.结果:决明子蛋白质最佳提取工艺为KH2PO4-NaOH缓冲液浓度为50 mmol/L,pH值为8.0,液固液比为1:50,提取12h.影响决明子蛋白提取各因素的主次关系为:缓冲液浓度＞pH值＞固液比＞提取时间.其中缓冲液浓度和pH值的影响迭显著性水平.优化参数下提取的决明子粗蛋白含量为93.0%,提取率为94.2%,体外消化率为97.6%.结论:决明子是含蛋白质较高的植物种子,其蛋白质消化率较高,具有很好的开发前景.     

不同对虾中多酚氧化酶的提取比较及在虾体的分布研究

用丙酮法和匀浆浸提法分别对南美白对虾和中华管鞭虾进行多酚氧化酶的提取,结果表明,在同样的提取条件下,中华管鞭虾的酶活大于南美白对虾,匀浆浸提法提取的酶活比丙酮法高1.5倍;同时对中华管鞭虾的提取条件料液比、缓冲液pH、浸提时间、水浴温度做了研究,通过正交实验得到当料液比1∶4,缓冲液pH8,浸提时间3h,水浴温度45℃条件下提取的酶具有最大的活性;进一步研究中华管鞭虾的多酚氧化酶在虾体中的分布情况,得出虾头部位多酚氧化酶含量较高,虾尾和虾身含量较低。     

菠萝蜜果皮中果胶的提取工艺优化

通过单因素和正交试验研究水提温度、pH值、提取时间、料液比4个理化因素对菠萝蜜果皮中果胶得率的影响,确定菠萝蜜皮果胶提取的最佳工艺参数。结果表明,酸提取最佳工艺为提取温度95℃、pH1.0、提取时间2.5h、液料比30:1(水:物料,m/m),果胶得率为12.46%;超声波辅助提取最佳工艺为80℃、50min、液料比40:1、pH1.0、450W,果胶得率为13.17%。超声波辅助提取可提高果皮中果胶得率和缩短时间。     

The effect of heating conditions on polyphenol oxidase, proteases and melanosis in pre-cooked Pacific white shrimp during refrigerated storage

Pre-cooked Pacific white shrimp (Litopenaeus vannamei) is an important shrimp product. However melanosis, especially in the cephalothorax including carapace and internal organs, is more likely caused by the remaining polyphenol oxidase (PPO) after pre-cooking. Thus, PPO from carapace and proteases from hepatopancreas of Pacific white shrimp were characterised and the remaining activities of both enzymes were monitored in pre-cooked shrimp during storage at 4 °C. Based on activity staining using L-β-(3,4 dihydroxylphenyl) alanine as a substrate, PPO consisted of two isoforms with apparent molecular weight of 210 and 220 kDa. No difference in activity band was observed when analysed under reducing and non-reducing condition. Proteases from hepatopancreas were able to activate PPO to some degree. For the in vitro study, both enzymes were quite stable when heated at temperature up to 70 °C but the loss in activities increased with increasing heating time (0-120 s). When Pacific white shrimp were pre-cooked to obtain different core temperatures (50-90 °C), different PPO and protease activities were retained. Higher core temperatures were associated with lower PPO and protease activities, but higher cooking loss. When the shrimp were pre-cooked at 80 °C, the residual PPO and protease activities were 3.9% and 5.4%, respectively and cooking yield of 95.6% was obtained. The resulting pre-cooked shrimp possessed lower melanosis score during 7 days of storage at 4 °C. Thus, pre-cooking of shrimp to obtain a core temperature of 80 °C, with a holding time of 30 s, could prevent the severe cooking loss and lower melanosis during subsequent storage.     

Edible mushroom (Flammulina velutipes) extract inhibits melanosis in Kuruma shrimp (Marsupenaeus japonicus)

This study compared the potential of an aqueous extract of an edible mushroom (Flammulina velutipes) to prevent melanosis in cultured Kuruma shrimp (Marsupenaeus japonicus) with other antimelanosic compounds in vivo. The mushroom extract contained 9.1 mg/mL ergothioneine (ESH). Immersion of live full-grown shrimp in a 0.5% w/v solution of mushroom extract significantly reduced PPO activity in shrimp hemolymph. In addition, expression of the prophenoloxidase (proPO) gene decreased in hemocytes, suggesting that the extract blocked the activation of the proPO cascade. Consequently, the development of melanosis in the treated shrimp was significantly suppressed during ice storage. Treatment with a 0.05% w/v solution of sodium ascorbate and 4-hexyl-1,3-benzenediol had the same effect. In vitro experiments showed that ESH effectively inhibited PPO activity and activation of the proPO cascade in hemocyte lysate supernatant. This study suggests that in vivo application of F. velutipes mushroom extract is an effective natural alternative to synthetic antimelanosic agents to inhibit postmortem melanosis in shrimp. Practical Application: The extract of an edible mushroom (F. velutipes) containing ergothioneine can be a promising natural alternative to synthetic antimelanosic agents used to prevent postharvest melanosis in shrimp and other crustaceans. Furthermore, utilization of the mushroom trimmings could also help address the growing concerns on the disposal of such agricultural wastes and instead use it into a novel purpose as a source of antimelanosic and antioxidants for food and industrial application.     

Effects of freeze-thaw cycles of Pacific white shrimp (Litopenaeus vannamei) subjected to radio frequency tempering on melanosis and quality

This study was conducted to investigate the effects of different tempering methods and freeze-thaw cycles on melanosis and quality parameters of pacific white shrimp. Frozen pacific white shrimps tempered with radio frequency tempering (RFT) were compared to that in water tempering (WT) and refrigerator tempering (RT) in terms of temper loss, total volatile base nitrogen (TVBN), polyphenol oxidase (PPO) activity, melanosis, total sulfhydryl contents, differential scanning calorimetry (DSC), and texture properties after 0, 1, 3, 5 freeze-thaw cycles. Results showed that crushed ice was effective as an effective surrounding medium for six layers of frozen shrimp reaching −2 °C within 6 min in RFT. For quality attributes, the temper loss of samples tempered with radio frequency is lower than that of RT and WT after all freeze-thaw cycles, and RFT resulted in the lowest TVBN value (9.17 mgN/100 g) of shrimps after the 5th freeze-thaw cycle. The PPO activity and melanosis of samples increased as the number of freeze-thaw cycles increased, and RFT effectively inhibited the development of melanosis. After the 3rd freeze-thaw cycle, the enthalpy change (△H) and the sulfhydryl content (0.16 mmol/gprot) of radio frequency tempered samples was significantly higher (p < 0.05) than that of WT and RT. RFT retained the hardness and chewiness of shrimp samples in all freeze-thaw cycles. Therefore, RFT effectively inhibited melanosis and reduce protein oxidation in Pacific white shrimp during freeze-thaw cycles with its fast and uniform heating characteristics.     

Inhibition of melanosis formation in Pacific white shrimp by the extract of lead (Leucaena leucocephala) seed

Lead (Leucaena leucocephala) seed extract was prepared using distilled water as a medium. An extraction yield of 26.16 g/100 g of seed was obtained after extraction at room temperature for 12 h. Total phenolic and mimosine contents in the lead seed extract powder (LSEP) were 17.4 g GAE/100 g and 8.8 g/100 g, respectively. LSEP at different concentrations (0.05%, 0.1%, 0.25%, 0.5%, and 1%, w/v) showed inhibitory activity towards polyphenoloxidase (PPO) of Pacific white shrimp in a dose dependent manner. When the whole Pacific white shrimp were treated with 0.25% and 0.5% (w/v) LSEP, the shrimp treated with 0.5% LSEP had the lower melanosis score throughout the storage of 12 days and showed a higher score for colour and odour, as well as overall likeness, compared with the control (without treatment) and 1.25% sodium metabisulphite treated samples at day 12 (P < 0.05). Meat of shrimps treated with LSEP at both levels had the increase in mimosine content up to 8 days, suggesting the migration of mimosine into shrimp muscle during extended storage. Therefore, 0.5% LSEP can be used as a novel melanosis inhibitor for Pacific white shrimp.     

Application of ergothioneine-rich extract from an edible mushroom Flammulina velutipes for melanosis prevention in shrimp,Penaeus monodon and Litopenaeus vannamei

This report investigated the melanosis-inhibiting properties of crude water soluble extract from the base and fruiting body waste of an edible mushroom (Flammulina velutipes) containing 9.13 ± 2.03 mg/mL 2-thiol-l-histidine-betaine (ergothioneine, ESH). Immersion of live full-grown black tiger shrimp (Penaeus monodon) and Pacific white shrimp (Litopenaeus vannamei) in a 0.5% w/v solution of mushroom extract for 1 h significantly decreased polyphenoloxidase (PPO) activity in shrimp hemolymph and expression of the prophenoloxidase (proPO) gene in hemocytes. Consequently, the development of melanosis in the treated shrimp during ice storage was prevented. Treatment with a 0.05% w/v solution of sodium sulfite and 4-hexyl-1,3-benzenediol had a similar effect. Enzyme assays showed that ESH is a non-competitive inhibitor. It is proposed that ESH possibly interacts directly with Cu²⁺ at the putative binding sites of PPO and proPO, based on copper-chelating activity analyses, thus preventing melanosis in the shrimp. This study indicated that application of ESH-rich F. velutipes mushroom extract from trimming waste is an effective natural alternative to synthetic melanosis-inhibiting agents to prevent postmortem melanosis in shrimp.     

Effect of cinnamaldehyde on melanosis and spoilage of Pacific white shrimp (Litopenaeus vannamei) during storage

BACKGROUND: Shrimp is a very perishable product and postmortem changes occur rapidly. Sulfiting agents were once and are still widely used as a preservative in the shrimp industry. However, the application of sulfite in shrimp may pose a risk to human health. Thus development of a natural preservative as a sulfite alternative to extend the shelf life of Pacific white shrimp is urgently needed. RESULTS: The effects of cinnamaldehyde essential oil (1 and 5 g kg^?1) on the shelf life of Pacific white shrimp stored at 4 °C were investigated. As the concentration of cinnamaldehyde increased, residual polyphenoloxidase (PPO) enzyme activity decreased. Kinetic analysis showed that cinnamaldehyde was a noncompetitive inhibitor for the oxidation of L ‐DOPA (L ‐3,4‐dihydroxyphenylalanine) by PPO of Pacific white shrimp. Based on this study, shrimp treated with 5 g kg^?1 cinnamaldehyde possessed the lowest aerobic plate count, total volatile basic nitrogen, and pH values in all treatments after 10 days of storage. According to the results of L*, cinnamaldehyde showed inhibitory activity toward the formation of melanosis. CONCLUSION: Treatment with cinnamaldehyde could improve the sensory properties and extend the shelf life of Pacific white shrimp to 8 days. Therefore, cinnamaldehyde could be used as a promising natural preservative for inhibiting melanosis and preventing the growth of microbes during the chilled storage of Pacific white shrimp. Copyright © 2012 Society of Chemical Industry     

Effect of pyrophosphate and 4-hexylresorcinol pretreatment on quality of refrigerated white shrimp (Litopenaeus vannamei) kept under modified atmosphere packaging

ABSTRACT:  The effect of pretreatment with pyrophosphate and 4-hexylresorcinol in combination with modified atmosphere packaging (MAP) (80% CO₂, 10% O₂, 10% N₂, or 80% CO₂, 20% N₂) on the quality of white shrimp during storage at 4 °C was investigated. Shrimp pretreated with 2% pyrophosphate and 0.25% 4-hexylresorcinol and stored under MAP showed the lower microbiological and chemical deteriorations as evidenced by delayed microbial growth as well as lower trimethylamine (TMA) and total volatile base nitrogen (TVB) production ( P < 0.05). Additionally, the growth of coliforms was inhibited effectively. White shrimp pretreated with 4-hexylresorcinol had the lower melanosis throughout the storage compared with those without treatment ( P < 0.05). This was associated with the lowered polyphenol oxidase (PPO) activity in shrimp treated with 4-hexylresorcinol. Therefore, the effective retardation of microbiological and chemical deterioration of white shrimp stored under MAP with the decrease in melanosis could be achieved by pretreatment of the shrimp with pyrophosphate and 4-hexylresorcinol. Furthermore, decapitation could be another means to lower the microbial load and melanosis in white shrimp, particularly those stored under MAP.     

Sulfur-Containing Compounds Heated under Alkaline Condition: Antibrowning, Antioxidative Activities, and Their Effect on Quality of Shrimp during Iced Storage

ABSTRACT:  Sulfur-containing compounds heated under alkaline condition (pH 10) were determined for inhibitory activity toward polyphenol oxidase (PPO) from Pacific white shrimp ( Litopenaeus vannamei ) and for antioxidative activity. Cysteine and glutathione (GSH) (20 mM) heated at 100 °C at pH 10 strongly inhibited PPO activity. Heated alkaline cysteine showed the greater 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging activity, copper-chelating activity and reducing power than cysteine and glucose–cysteine Maillard reaction products ( P  < 0.05). Effect of heated alkaline cysteine at different concentrations (0, 20, and 100 mM) on the quality changes of Pacific white shrimp during iced storage was investigated. Shrimp treated with 100 mM heated alkaline cysteine had the lowest melanosis score, thiobarbituric acid-reactive substances (TBARS) value and total viable count (TVC), compared with those without treatment and treated with 20 mM heated alkaline cysteine, throughout the storage of 12 d ( P  < 0.05). Therefore, heated alkaline cysteine can be used as a novel additive to retard melanosis and extend the quality of postmortem shrimp.     

Effect of Green Tea Extract in Combination with Ascorbic Acid on the Retardation of Melanosis and Quality Changes of Pacific White Shrimp During Iced Storage

Melanosis and quality changes of Pacific white shrimp (Litopenaeus vannamei) treated with 0.1% green tea extract (GTE) in combination with ascorbic acid (AA) at different levels (0%, 0.005%, and 0.01%) were monitored during iced storage of 12?days. Based on in vitro study, 0.1% GTE inhibited polyphenoloxidase (PPO) from cephalothorax of Pacific white shrimp by 60.2%. Nevertheless, 0.1% GTE in combination with 0.01% AA exhibited the greater PPO inhibitory activity (93.0%) (P?<?0.05). When shrimp treated with 0.1% GTE in combination with AA (0.005 or 0.01%; GTE + AA), the increase in psychrophilic bacteria and spoilage microorganisms including H₂S- producing bacteria and enterobacteriaceae were retarded to a higher extent, in comparison with the control and those treated with 1.25% sodium metabisulfite (SMS; P?<?0.05). The coincidental lowered rates of increase in pH, total volatile base content and thiobarbituric acid reactive substances were obtained in the shrimp treated with GTE + AA (P?<?0.05). Additionally, shrimp treated with GTE + AA had the lower melanosis score but higher score for color, odor, taste, flavor, and overall likeness, compared with the control and those treated with SMS (P?<?0.05). Generally, AA at levels of 0.005% and 0.01% showed a similar synergist effect with GTE on both melanosis inhibition as well as retardation of quality loss of shrimp.     

虾体多酚氧化酶特性及其抑制技术研究进展

多酚氧化酶是虾体中普遍存在的一种含铜金属蛋白,它是虾体发生黑变的主要原因,也是虾保鲜品质控制中亟待解决的技术关键。研究多酚氧化酶的酶学特性,抑制酶促褐变,对于提高虾的食用价值和商品价值至关重要。本文综合介绍了多酚氧化酶的虾体分布、活性特征、分离纯化、分析测定、黑变机理及其抑制方法。