响应面法优化制备牡蛎短肽工艺

【目的】探究前处理方式对酶解效果的影响,优化牡蛎短肽制备工艺。【方法】在确定高效酶种类和前处理方式的基础上,以氮回收率、短肽得率为指标对酶解时间、料液比、酶解温度、加酶量进行单因素试验,再利用响应面设计建立数学模型,以短肽得率为响应值,进行4因素3水平的响应面分析。【结果与结论】牡蛎经80℃热处理10 min后使用动物蛋白酶的酶解效果最佳。响应面结果显示,最佳酶解工艺为料液比(g/m L)1∶3.9、温度47℃、加酶量3 300 U/g、自然pH(6.5)、酶解3 h,其短肽得率为(58.53±1.20)%,比原酶解工艺提高24.8%。     

对虾蛋白自溶制备ACE抑制肽的工艺条件优化

以凡纳滨对虾为原料，以ACE抑制率为指标，利用响应面法对虾肉蛋白自溶制备ACE抑制肽的工艺条件进行了优化，即在酶解条件（pH值、温度、虾头虾肉质量比）和ACE（Angiotensin I-converting Enzyme，ACE）抑制率之间建立了数学模型Y=23．59—0．21X1＋0．84X2＋0．85X3—0．71X1^2-0．94X2^2-1．06X3^2＋0．088X1X2—0．46X1X3-0．87X2X3。分析表明，在3个因素中，虾头与虾肉比例对ACE抑制率的影响最为显著。优化后的工艺参数为：pH7．35，温度57．2℃，虾头与虾肉比例为1：1。根据回归方程的预测结果，反应时间为4h，其ACE抑制率达41．9％。     

对虾蛋白自溶制备ACE抑制肽的工艺条件优化

以凡纳滨对虾为原料,以ACE抑制率为指标,利用响应面法对虾肉蛋白自溶制备ACE抑制肽的工艺条件进行了优化,即在酶解条件(pH值、温度、虾头虾肉质量比)和ACE(Angiotensin I-converting Enzyme,ACE)抑制率之间建立了数学模型Y=23.59-0.21X1+0.84X2+0.85X3-0.71X12-0.94X22-1.06X32+0.088X1X2-0.46X1X3-0.87X2X3。分析表明,在3个因素中,虾头与虾肉比例对ACE抑制率的影响最为显著。优化后的工艺参数为:pH7.35,温度57.2℃,虾头与虾肉比例为1∶1。根据回归方程的预测结果,反应时间为4 h,其ACE抑制率达41.9%。     

响应面法优化波吉卵囊藻多糖提取条件

【目的】探究波吉卵囊藻(Oocystis borgei)多糖提取的最优条件。【方法】通过单因素和响应面实验,考察不同提取温度、氢氧化钠浓度、提取时间对波吉卵囊藻多糖提取率的影响。【结果】各因素对波吉卵囊藻多糖提取率的影响显著性表现为氢氧化钠浓度提取温度提取时间;最佳提取条件:提取温度58.5℃、氢氧化钠浓度0.305mol/L、提取时间68 min,此时提多糖取率为11.86%,与模型预测值11.93%接近。【结论】采用响应面法建立的模型可以较好地预测提取温度、氢氧化钠浓度和提取时间对波吉卵囊藻多糖的提取率的影响。     

酶法制备海蛰氨基多糖工艺的优化

海蜇（Rhopilema esculentum）属于腔肠动物门（Coelentera）、钵水母纲（Scyphozoa）、根口水母目（Rhizostomeae）、根水母科（Rhizostoidae）、海蜇属（Rhopilema），是“海产八珍”之一，俗称水母、面海蛰，为我国沿海大型水生生物和重要水产资源。     

酶法制备海蛰氨基多糖工艺的优化

海蜇(Rhopilema esculentum)属于腔肠动物门(Coelentera)、钵水母纲(Scyphozoa)、根口水母目(Rhizostomeae)、根水母科(Rhizostoidae)、海蜇属(Rhopilema),是“海产八珍”之一,俗称水母、面海蛰,为我国沿海大型水生生物和重要水产资源。海蛰味道鲜美、风味独特,深受人们喜爱,并具有多种药理功能[1,2]。《本草纲目》记载,海蛰可入药,“主治妇人劳损,积血带下,小儿风疾丹毒,烫火伤”。《医林纂要》言其“补心益肺,滋阴化痰,去结核,行邪湿,解酒醒渴,止嗽除烦”。《归砚录》言“海蜇,妙药也。宣气化淤,消痰行食而不伤正气”。著名中药古方“…     

酶解蓝圆鲹鱼肉蛋白制备小分子肽的工艺研究

以蓝圆鲹为原料,酶解蓝圆鲹鱼肉蛋白制备小分子肽。运用响应面试验设计优化蓝圆鲹蛋白酶解的工艺条件,探讨了酶种类和添加量、液固比、酶解温度、酶解时间对多肽提取率的影响。结果表明:选用木瓜蛋白酶,添加量为0.10%,液固比2∶1,酶解温度54.4℃,酶解时间3.18 h下,多肽提取率为32.35%。     

基于模糊数学评价法优化凡纳滨对虾真空微波干燥工艺

【目的】探求真空微波干燥凡纳滨对虾的最佳工艺条件,以期为开发凡纳滨对虾即食食品提供技术参考。【方法】以新鲜凡纳滨对虾为原料,模糊数学感官评分为主要考察指标,在单因素试验基础上,选择载装量A、功率B、真空度C为自变量,利用响应面法和模糊数学评价法对真空微波干燥工艺进行优化。【结果】以感官评定为基础对真空微波干燥凡纳滨对虾的品质进行权重分析,得到权重集K=(色泽0.335,滋味0.305,香气0.215,组织质地0.145)。建立二次多项回归方程:Y=84.588+1.084A+1.409B+1.700C-7.353A²-8.793B²-13.860C²,显示载装量A、功率B、真空度C对感官评分有极显著影响(P<0.01)。通过响应面分析,得到真空微波干燥凡纳滨对虾的最佳工艺条件:载装量75 g,加热功率400 W,真空度0.08 MPa。在此条件下,真空微波干燥凡纳滨对虾品质良好,感官评分为(85.6±0.07)分,与预测值85.92分误差较小。【结论】响应面法优化凡纳滨对虾真空微波干燥工艺具有操作简单、优化条件精确、结果显示直观等优点;建立的回归方程能反映装载量等三个因素对指标的影响规律,并可进行实际预测。     

基于粗糙集理论与响应面法的老黏土膨胀敏感性分析方法

评价老黏土膨胀性等级时出现的误差主要源于多因素在参数统计上的不确定性,对此提出老黏土膨胀敏感性分析方法。结合粗糙集理论分析各因素的敏感性并确立主导因素,进而构建老黏土膨胀性等级的多因素定量评价模型,随后通过响应面法,分析主导因素间参数不确定性的交互作用对评价结果的影响。襄阳城区老黏土研究表明,自由膨胀率和体缩率为影响评价结果的主导与次主导因素,二者参数变化产生的交互作用对膨胀性评价等级影响较显著。当体缩率统计值较大时,随着自由膨胀率的上升,膨胀性评价等级变幅较大,偏大的自由膨胀率将高估老黏土的膨胀性,致使评价过于保守,偏小的自由膨胀率则会对其低估,导致结果较不可信;而当体缩率统计值较小时,评价等级随自由膨胀率的上升变化不大,因此评价结果较为准确。所得结论可为老黏土膨胀性评价与分类工作提供参考。      

Purification and Characterization of a Novel Lipase from Antarctic Krill

Lipase from Antarctic krill,with a molecular weight of 71.27kDa,was purified with ammonium sulfate precipitation and a series of chromatographic separations over ion exchange(DEAE)and gel filtration columns(Sephacryl S-100),resulting in 5.2%recovery with a 22.4-fold purification ratio.The optimal pH and temperature for enzyme activity were 8.0 and 45℃,respectively.Purified lipase had Km and Vmax values of 3.27mmolL−1 and 2.4Umg−1,respectively,using p-nitrophenyl laurate as the substrate.Lipase activity was enhanced by adding Ca2+and Mg2+ions in the concentration ranges of 0–0.5mmolL−1 and 0–0.3mmolL−1,respectively,while the activity was inhibited by a further increase in these ion concentrations.Fe3+and Cu2+ions showed obvious inhibitory effects on enzyme activity,and the inhibition rates were 71.8%and 53.3%when the ion concentrations were 0.5mmolL−1.     

Purification of antimicrobial peptide from Antarctic Krill (Euphausia superba) and its function mechanism

The preliminary purification and antimicrobial mechanism of antimicrobial peptide from Antarctic Krill were studied in this paper. The results showed that the molecular weight range of antimicrobial polypeptide (CMCC-1) obtained by cation exchange chromatography was between 245-709D as detected by molecular sieve chromatography, and the minimum inhibition concentration (MIC) of CMCC-1 against Staphylococcus aureus was 5.0 mg mL^?1. The antimicrobial mechanism of CMCC-1 was studied with S. aureus as indicator bacterium. Compared with control group, the results of the experimental group in which S. aureus was treated with CMCC-1 were as follows: 1) CMCC-1 could inhibit cell division at logarithmic phase. 2) The protein and reducing sugar content, and the conductivity of culture medium increased, and the activity of alkaline phosphatase and β-galactosidase could be detected in the culture medium. 3) Observation under scanning electron microscope revealed that somatic morphology became irregular, and then somatic surface became coarse. The cell became much smaller, and most somatic cells gathered. The boundary between cells became dim and finally fused as a whole. 4) Observation under transmission electron microscope showed that the surface of S. aureus became rough and the reproducing ability was restrained. The cell wall became thin and the cytoplasm shrunk. Substances inside cell leaked out, which caused cells death. 5) SDS-PAGE analysis showed that some bands disappeared, and the residual bands became vague. 6) The genomic DNA electrophoresis results showed that the genomic DNA bands of S. aureus were not degraded but the brightness significantly reduced. Thus, it is supposed that CMCC-1 could destroy the cell wall and membrane of S. aureu, increase the cell membrane permeability and the leaking-out of intracellular substances, and thus cause the death of S. aureu.     

Optimization of Hydrolysis Conditions for the Isolation of Angiotensin-I Converting Enzyme(ACE) Inhibitory Peptides from Rhopilema hispidum

To obtain the maximum angiotensin-I converting enzyme(ACE) inhibitory activity, the protein hydrolysis conditions of the jellyfish Rhopilema hispidum were optimized using response surface methodology(RSM). Trypsin was selected to produce R. hispidum protein hydrolysates(RPH) with ACE inhibitory activity. The optimal parameters for producing protein hydrolysates with the highest ACE inhibitory activity were as follows: hydrolysis time 5 h, hydrolysis temperature 50℃, and the enzyme-to-substrate ratio 6%. Under these conditions, the ACE inhibitory rate of RPH could reach 64.28% ± 5.72%. In addition, RPH contained high levels of Gly, Glu, Pro, Ala, Asp and Arg, with a molecular weight distribution range of 0.32–6.84 kDa. The following three novel ACE inhibitory peptides were isolated and identified: Ile-Gly-Glu-Thr-Gly-Pro, Gly-Ala-Thr-Gly-Pro-Ala-Gly-Tyr-Val and Gly-AlaPhe-Gly-Pro-Gly-Gly-Leu-Val-Gly-Arg-Pro. The IC_(50) values of the ACE inhibitory activity of these three purified peptides were 19.07, 27.42 and 31.26 μmol L~(-1), respectively. These results suggested that proteins and peptides isolated from R. hispidum could be utilized as antihypertensive functional food sources.     

马氏珠母贝免疫活性肽的纯化与鉴定

为制备马氏珠母贝免疫活性肽,以马氏珠母贝全脏器为原料,利用体外模拟消化的方法对其进行酶解,然后通过超滤系统收集分子量小于3 ku的酶解液。采用细胞培养方法,以体外免疫活性为指标,利用Sephadex G-25凝胶柱层析、Capto Q强阴离子交换色谱等对马氏珠母贝酶解液进行分离纯化与筛选,获得2个具有免疫活性的肽段,利用电喷雾静电场离子阱串联质谱分别对2个活性肽序列进行测定。结果表明:一个肽的相对分子质量为245.0,肽序列为Ala-Arg/Pro-Met;另一个别肽的相对分子质量为274.0,肽序列为Val-Arg。在浓度0.3 mg/m L时,2种肽能显著促进脾淋巴细胞的增殖与提高巨噬细胞的吞噬能力,表明这2种肽均具有较好的免疫活性。     

罗非鱼鱼鳞胶制备降血压胶原肽的工艺及酶解物活性

以罗非鱼鱼鳞胶为原料,制备对血管紧张素转换酶(ACE)有抑制活性的降血压肽。以水解度(DH)及ACE抑制率为指标,筛选水解鱼鳞胶的最适酶种,并对其水解工艺条件进行优化;采用超滤技术对所制备的降血压肽进行分离纯化,比较不同组分的ACE抑制率,测定高ACE抑制率组分的氨基酸组成,并运用MALDI-TOF-MS法分析其分子质量分布。结果表明,复合蛋白酶是水解鱼鳞胶最适酶种,在50℃、pH7.5、料液质量体积比1∶5、酶与底物质量比(mE/mS)1.5%条件下,酶解6 h,酶解液的ACE抑制作用最强,其水解度DH 73%,ACE的抑制率76.15%。酶解液经超滤分离所获得的分子质量小于2 000 u的组分对ACE抑制率最高,达83.15%;该组分的氨基酸主要含有Gly和Pro,含量分别为22.04%和11.40%;该组分分子质量范围为1 300~2 000 u。