鲟鱼硫酸软骨素的制备工艺研究

以史氏鲟鱼软骨为原料,按碱提-酶解-乙醇沉淀工艺流程进行提取硫酸软骨素的各主要工序工艺的研究.通过单因素试验及正交试验,得出最佳碱提工艺为以两倍量6%的NaOH为溶剂,于40℃的水浴中搅拌提取4 h.通过正交试验,得出最佳酶解工艺条件是先将浸提液调节pH至8.6,加入胰酶0.5%,50℃保温水解3 h;后降温至40℃,调pH至5.7,加入胃蛋白酶0.3%,保温水解2h.通过单因素试验,得出最佳的乙醇沉淀工艺条件是将滤液pH调为6.5,加入95%的乙醇至醇沉浓度为70%,静置过夜.制备的硫酸软骨素经检测符合WS1-C3-0030-2000(硫酸软骨素口服片)和WS-10001-(HD-0892)-2002(硫酸软骨素注射品)标准的要求.     

大孔吸附树脂纯化海地瓜总皂苷的研究

本文研究了海地瓜总皂苷的纯化工艺,并采用3种大孔吸附树脂对海地瓜皂苷的吸附作用进行了比较分析,筛选出最佳的吸附树脂,并对该树脂对海地瓜皂苷的静态吸附和动态吸附效果进行测定。结果表明,筛选出的D101大孔树脂为最合适的树脂型号;以D101大孔树脂静态吸附条件进行纯化,获得皂苷纯度为51.4%;以D101大孔树脂对海地瓜粗提液进行动态吸附研究,获得皂苷纯度达71.3%。因此,D101大孔树脂能够有效地应用于海地瓜总皂苷的富集和纯化。     

碱液对牦牛软骨提取硫酸软骨素工艺的影响

以牦牛软骨为原料,采用先高温蒸煮,后加稀碱与酶解相结合的方法提取硫酸软骨素。影响提取效果的因素主要有碱液浓度、碱提温度、碱提时间等,试验采用三因素三水平正交试验设计,对硫酸软骨素提取过程中碱液的影响进行研究,试验结果表明:碱液浓度5%,碱提取温度40℃,碱提取时间6h为牦牛硫酸软骨素提取的最佳碱液提取工艺参数,出品率可达10.03%,有效成分含量达91.67%。     

海带岩藻聚糖硫酸酯电渗析分离工艺

采用电渗析法对盐沉后的海带酶解液中岩藻聚糖硫酸酯进行分离,探索工业化分离生产岩藻聚糖硫酸酯的方法,以解决目前乙醇沉淀法存在的安全性和高成本等问题。以脱盐率、糖保留率为检测指标,采用响应面法优化了电渗析工艺参数。结果显示,在操作电压80 V、料液流量3.2 L/min、固形物浓度10 mg/ml、p H=5的条件下,电渗析连续循环3次,海带酶液解干基中多糖含量由40.17%提高至47.11%,硫酸根含量由3.36%提高至4.97%。电渗析后海带酶解液干基中的金属离子含量为Ca 0.13 mg/g、Mg 16.04μg/g、Cu 0.29μg/g、Se 11.7μg/g、Cr 8.06μg/g,Pb未检出,较电渗析前均有降低。电渗析对脱除褐藻胶的海带酶解液进行脱盐纯化处理,可有效地分离纯化出岩藻聚糖硫酸酯。     

大孔吸附树脂分离纯化羊栖菜多酚工艺研究

通过树脂吸附-解吸实验分析,对羊栖菜多酚大孔树脂柱层析纯化工艺进行研究。结果表明:XDA-7型大孔树脂表现出良好的吸附与解吸性能;羊栖菜多酚柱层析纯化工艺条件为:上样浓度3.20 mg/m L,上样流速1 BV/h,洗脱剂为60%乙醇溶液,洗脱流速2 BV/h,在此条件下纯化制备的羊栖菜多酚样品纯度为61.86%。     

提取鲟鱼硫酸软骨素技术

<正>技术简介:1.本技术采用季铵盐-乙醇沉淀法分离纯化鲟鱼硫酸软骨素,探究此纯化方法的最优条件,研究结果表明:季铵盐浓度1%、作用温度20℃、静置放置1.5h,乙醇沉淀浓度为75%、溶液p H值为7。此工艺所得鲟鱼头骨、脊骨硫酸软骨素纯品的产率分别为19.5%、22%,纯度分别为94.8%、95.32%,蛋白含量为0.287%、0.267%。2.对所得纯品与鲨鱼硫酸软骨素纯品进行质量比较,均符合标准;紫外扫描结果在260nm、280nm无吸收峰     

影响淡水鱼头酶解的因素研究

选用中性蛋白酶和碱性蛋白酶对草鱼鱼头进行水解,单因素分析和正交实验L9(34)相结合,对影响其酶解条件的因素进行了探讨.结果表明:碱性蛋白酶水解效果较中性蛋白酶好,其最佳酶解条件为酶用量0.018%、温度53℃、pH值7.6、水解5h,在此条件下酶解液中游离氨基酸可达1.25mg/ml.并分析了两种酶在最佳酶解条件下,可溶性蛋白质含量的变化.     

碱液浸提法制备硫酸软骨素的工艺优化

采用碱液浸提法对鱿鱼软骨中硫酸软骨素的提取工艺进行研究。以硫酸软骨素得率为评价指标,通过对影响硫酸软骨素得率的因素进行单因素试验和响应面优化试验确定最佳工艺参数。结果表明,碱液浸提的最佳工艺条件为：料液比1∶8( g/mL)、浸提温度50℃、浸提时间5．3 h, NaOH浓度3%。在此条件下,鱿鱼软骨中硫酸软骨素得率为62．89%。     

年产2t硫酸软骨素车间设计初探

选择鲟鱼鱼糜加工过程中废弃的鱼骨为原料,采用稀碱提取-酶解-醇沉工艺,获得具有多种药用价值的硫酸软骨素,并进行年产2t硫酸软骨素车间设计。按年产2t硫酸软骨素车间进行工艺流程、设备选型、物料衡算、关键点控制等方面进行设计,并绘制工艺流程图和车间平面图。设计为硫酸软骨素的生产提供参考依据。     

贻贝粉酶解液中系统提取分离氨基酸

本文报道了以脱脂贻贝粉为原料，采用酶解法从其酶解液中分离，提取及纯化复合氨基酸和六种单一氨基酸的工艺。确定了贻贝粉以胃蛋白酶、枯草杆菌中性蛋白酶和胰蛋白酶联合的酶解工艺，采用超滤方法去除酶解液中未水解的蛋白质等大分子物质及进行脱色处理。将超滤液通过７０１＃［ＣＯ＝］型碱性阴离子交换树脂及［Ｈ＋］型１×２５强酸性阳离子交换树脂脱盐，得到的脱盐液经真空浓缩、结晶、洗涤和烘干得复合氨基酸粉末，得率为２０％左右（相对子贻贝粉，ｗ／ｗ，下同）。将超滤液通过活性炭柱吸附苯丙氨酸和酪氨酸，再用洗脱液洗下，经浓缩、结晶、洗涤和烘干得单一氨基酸成品，Ｌ－苯丙氨酸得率为２．４５％，Ｌ－酪氨酸得率为１．２１％。活性炭柱的通过液经７３２强酸性阳离子交换树脂柱及７１７强碱性阴离子交换树脂柱组吸附、分离、洗脱得纯 Ｌ－精氨酸组分、纯 Ｌ－谷氨酸组分、Ｌ－亮氨酸及Ｌ－缬氨酸组分。再经赶氨、浓缩、结晶、洗涤和烘干得成品，得率分别为３．０１％、１．０２％、１． ４６％及 １． ３４％。单一纯化的氨基酸产品，经氨基酸自动分析仪定性、定量测定及红外光谱分析和通过自动指示旋光仪测定其比旋度进行鉴定，证实确为本物质。复合氨基酸产品中游离氨基酸含量为９０．     

An Effective Method for Cadmium Removal from Scallop By-product Enzymatic Hydrolysate

An effective method to remove cadmium (Cd) from scallop by-product enzymatic hydrolysate (SBEH) was developed, and the Cd bonding sites were studied. Cation-exchange resin, anion-exchange resin, activated charcoal, and zeolite were tested. The result showed that strong acid polystyrene cation-exchange resin was the most suitable sorption material for the removal of Cd. The optimal conditions were as follows: SBEH concentration at 17.07 mg/mL, pH value at 4.92, and adsorption time with 11.73 min, while 4 g resin was added in 50 mL solution at 30°C. The experimentally measured removal rate was 95.38% under these conditions. The Cd element has an intense bonding pattern with macromolecules, and more than half of the Cd element exists as positively charged ion in the SBEH. The cation-exchange resin removed Cd by dumping the macromolecules associated with the Cd element. Infrared (IR) spectra suggested that the sulfate ester groups and carboxyl groups in the macromolecules were binding sites of the Cd element in SBEH. These results provide a useful method for Cd removal from SBEH that potentially can be used for industrial processing.     

酶解扇贝裙边制备复合氨基酸螯合钙的研究

扇贝裙边富含蛋白质、脂质等营养成分。为了高值化利用扇贝裙边，本研究选用中性蛋白酶、动物蛋白酶、风味蛋白酶、木瓜蛋白酶和酸性蛋白酶5种酶，以酶解液中游离氨基酸态氮为考察指标，对扇贝裙边进行酶解工艺条件探讨。首先，将5种酶制成复合蛋白酶进行正交实验，确定最佳酶解时间、温度、pH及加酶量，经检验，氨基酸转化率为77%；然后，通过实验确定CaCl2为最适钙源，以贝壳为原料，通过水飞法和酸法转化可制得贝壳源CaCl2。将扇贝裙边酶解液中复合氨基酸与来源于贝壳的钙螯合制备复合氨基酸螯合钙，以正交实验筛选出最佳螯合条件。经检验，该螯合反应螯合率达92%。     

Characterization and distribution of undersulfated chondroitin sulfate and chondroitin in leptocephalous larvae of elopomorph fishes

Undersulfated chondroitin sulfate (i.e., chondroitin sulfate with sulfate: hexosamine molar ratios ranging from 0.06–0.17) has been identified as a principal glycosaminoglycan (GAG) in whole-body extracts of leptocephalous larvae of four species of marine teleost fishes, representing two orders and three families. These include the eels (Anguilliformes)Ariosoma balearicum (Congridae),Rhechias dubia (Congridae) and Ophichthus sp. (Ophichthidae) and the ladyfish (Elopiformes: Elopidae: Elops saurus). A second GAG found in relatively large amounts in A. balearicum has been identified as chondroitin. Chondroitin and undersulfated chondroitin sulfate formed distinctive precipitates during extraction and were easily separated. Undersulfated chondroitin sulfate also was extracted and purified from early metamorphosing bonefish (Albuliformes: Albulidae: Albula sp.) leptocephali, where it is a minor GAG component, and was shown to be similar in composition to the compounds obtained from eels and ladyfish.     

Purification and physicochemical properties of deoxyribonuclease from pyloric caeca of Atlantic cod (Gadus morhuaL.)

A deoxyribonuclease (DNase) of pancreatic origin has been purified from extracts of the pyloric caeca from Atlantic cod (Gadus morhua L.). The crude extract was prepared by mincing frozen caeca tissue in equal volumes of buffer. The enzyme was isolated from the supernatant after streptomycin sulfate precipitation and centrifugation. The purification scheme further included chromatography on Q-Sepharose Fast Flow and hydroxyapatite columns. Affinity adsorption chromatography of the hydroxyapatite fraction on 8-(6-aminohexyl)-amino-5′-AMP-Sepharose, revealed an apparently homogeneous protein with molecular weight of 35,000 Da as judged by NaDodSO₄-PAGE. In sum a 644-fold enzymatic enrichment and 3.5% total enzyme recovery was achieved. The cod enzyme resembles DNase I-type enzymes with an alkaline pH activity optimum and shows dependency for Mg²⁺. The pI of the enzyme is 6.5 as determined by isoelectric focusing and DNase-zymography. Our findings suggest that the nuclease is a member of the cod's digestive enzymes secreted from the connective tissue surrounding the caeca.     

具有结合胆酸盐作用卵形鲳鲹蛋白酶解物的制备和分子量分布研究

以卵形鲳鲹(Trachinotus ovatus)鱼肉为原材料,以酶解产物与胆酸盐的体外结合率为指标,从5种常用食品用蛋白酶中筛选最优蛋白酶。进而采用正交法优化酶解条件,以提高酶解产物的胆酸盐结合率,并采用GPC法分析产物中蛋白肽的分子量分布情况。结果表明,胰蛋白酶酶解产物(100 mg·m L~(-1))与胆酸钠、甘氨胆酸钠和牛磺胆酸钠等3种胆酸盐的体外结合能力均好于其他4种蛋白酶,结合率分别达到42.1%、33.5%和30.1%,分别相当于降血脂药物考来烯胺散(20 mg·m L~(-1))的78.3%、74.4%和76.8%;经过正交试验优化的制备卵形鲳鲹蛋白肽的工艺条件为酶解时间2 h,加酶量2 000 U·g~(-1),料液比1∶4(g·m L~(-1)、p H 8、37℃)。该条件下酶解产物(20 mg·m L~(-1))对甘氨胆酸钠体外结合率达到同浓度阳性对照物(考来烯胺散)的48.3%,酶解产物中相对分子量3 k D以下的肽类组分占总蛋白水解物的77.30%。     

Simple Technologies for Converting Rest Raw Materials of Atlantic Salmon (Salmo salar) into High-Quality,Valuable, and Tasty Feed Ingredients

Rest raw materials as viscera, heads, and frames from farmed Atlantic salmon (Salmo salar) were hydrolyzed with the use of endogenous enzymes and the commercial enzymes Protamex and a mixture of Papain and Bromelain. Composition of the prehydrolysis mixture clearly influenced the quality of final hydrolysate and process kinetics. An increased proportion of viscera increased the amount of endogenous enzymes, which influenced hydrolysis kinetics, the extent of hydrolysis, and increased bitterness. Commercial enzymes, in addition to endogenous enzymes, are not always more yield efficient or economically beneficial but can be used to improve the taste and to ease the separation of oil from the hydrolysate fraction. Hydrolysis of denatured proteins in rest raw materials was hardly detectable. Optimum temperature should therefore be selected to avoid protein denaturation and simultaneously maintain high enzymatic activity. Hydrolysates preferably contain a low concentration of oil, and this work shows that high-quality oil can be separated before hydrolysis by mild thermal treatment. The previous separation of oil did not influence hydrolysate yield and decreased the concentration of lipids in the final hydrolysate. The initial separation of oil also increased productivity of the hydrolysis reactor, due to the reduction of hydrolysis volume.     

Production of Interesting Peptide Fractions by Enzymatic Hydrolysis of Tuna Dark Muscle By-Product Using Alcalase

ABSTRACT

A protein hydrolysate was prepared from proteins of tuna dark muscle by-product. The hydrolysis conditions (time, temperature, pH, and enzyme concentration) using Alcalase was optimized by response surface methodology (RSM). The regression coefficient close to 1.0, observed during experimental and validation runs, indicated the validity of the model. The hydrolysate produced under the optimum conditions determined by RSM has a low rate of peptide fraction of molecular weight of 4–1 kDa. Meanwhile, the results obtained by hydrolysis under optimal conditions determined by a complementary study (temperature 55°C, time 60 min, 1% enzyme concentration, and pH 8.5) show that the hydrolysate produced has a height rate of the peptide fraction of molecular weight of 4–1 kDa. The amino acid composition of the protein hydrolysate prepared proved to have the potential for application as an ingredient in balanced fish diets and as a source of nitrogen in microbial growth media.     

pH和色谱柱对日本鳗鲡肝脏抗菌肽分离纯化效果的影响及抗菌活性检测

为了能够快捷地分离纯化出单一的抗菌肽,实验采用不同pH值(离子交换流动相pH3.0、4.0、5.0;反相液相层析流动相pH 2.0、4.5、7.0)缓冲液,不同色谱柱(离子交换和凝胶过滤层析柱分别与反相液相层析柱)联用对日本鳗鲡肝脏抗菌肽分离纯化效果进行了比较,并分析了琼脂板扩散法和微孔液体培养法检测抗菌活性的优缺点.结果显示:pH 4.0缓冲液为最佳离子交换流动相,蛋白提取率为16.43％,得到2个洗脱峰;反相液相层析3种pH缓冲液分离效果均不理想,凝胶过滤层析柱与反相液相层析柱联用能明显改善日本鳗鲡肝脏抗菌肽分离纯化效果,洗脱峰数量较多,峰形单一且尖锐狭窄,基线平而低.琼脂板扩散法检测抗菌活性操作简单,实验结果直观,但所需蛋白量较多,可用于蛋白粗提物抗菌活性的检测;微孔液体培养法检测抗菌活性灵敏,所需蛋白较少,可用于后期色谱分离纯化蛋白抗菌活性的检测.