猪血药用物质基础研究(Ⅱ)- 酶种类的选择

通过体外模拟体内条件,选取主要消化酶胃蛋白酶、胰蛋白酶对猪血脱色液进行酶解,以高效酶木瓜蛋白酶和1394酶作为对照,结合酶解后多肽总含量以及<3 kDa的多肽得率进行酶解效果的综合评价,得到胃蛋白酶为最佳酶解酶。     

不同酶对草鱼蛋白功能特性及风味成分的影响

通过蛋白酶对草鱼进行酶解,对其酶解产物的功能特性和酶解液的风味成分进行研究。结果显示,风味蛋白酶、胰蛋白酶酶解产物的溶解性、热稳定性均优于胃蛋白酶酶解产物(P0.05)。经酶处理后鱼肉酶解液鲜甜味氨基酸占比增加,苦味氨基酸占比减少;利用SPME-GC-MS共检测出83种挥发性成分,以醛酮类和醇类为主,酶解处理后醇类的相对含量显著减少,醛酮类的相对含量则显著增加,酶解液具有独特风味。经胰蛋白酶、风味蛋白酶酶解后的草鱼肉酶解产物及酶解液均可作为食品基料应用于加工中。     

从米糠中提取降血管紧张素转换酶抑制剂

采用盐析法提取米糠蛋白,分别选用胃蛋白酶、胰蛋白酶、木瓜蛋白酶等单独或联合水解米糠蛋白,以从酶解米糠蛋白中分离获得具有血管紧张素转换酶（ACE）抑制活性的短肽组分。经Sephadex G-15凝胶层析和SP—Sephadex C-25离子交换层析分离各酶解组分,并检测各组分的ACE抑制活性。结果表明,米糠蛋白的酶解分离产物中含有较强ACE抑制活性的组分,其中经胃蛋白酶和胰蛋白酶共同水解时得到的小分子量寡肽组分的抑制活性最强,为后续对其进行结构分析奠定了基础。     

酶法提取金乌贼墨汁中黑色素的工艺条件研究

以金乌贼墨汁为原料,采用单因素试验,对胰蛋白酶,木瓜蛋白酶,胃蛋白酶,中性蛋白酶和碱性蛋白酶等五种酶提取黑色素的效果进行比较分析,筛选出水解效果较好的碱性蛋白酶。并通过正交试验,优化了碱性蛋白酶酶解金乌贼墨汁的最佳工艺,实验结果表明,在底物浓度2%、水解温度50℃、pH值为7.4,加酶量4200U/g和水解8h的条件下进行水解的效果较好。     

蚯蚓外源酶与内源酶酶解产物分析

本试验采用了枯草杆菌(Bacillus subtilis)AS1.398枯草杆菌蛋白酶和自溶制备蚯蚓肽,通过前期单因素试验,设计正交试验L9(34),以氨基氮数,多肽浓度为衡量指标,对最佳酶解条件进行筛选研究,并分析了酶解液氨基酸组成和相对分子量的分布。结果表明:AS1.398枯草杆菌蛋白酶最佳酶解条为pH 6.5、酶浓度1%、温度50℃、反应时间8 h,在此条件下,酶解蚯蚓蛋白的每100 mL水解液氨基氮数可以达到16.55 mmol,多肽浓度达9.22 mg/mL;自溶酶解蚯蚓的最佳条件pH 7.0、底物浓度1∶2、温度50℃、反应时间6 h,在此条件下,酶解蚯蚓蛋白的每100 mL水解液氨基氮数可以达到21.55 mmol,多肽浓度达11.65 mg/mL;二者酶解产物分子量大部分是在5000以下的多肽、小肽及氨基酸的混合物,其中分子量在300以下占了80%,氨基酸组成平衡,含量丰富,可用来制取新型氨基酸微量元素及小肽添加剂。通过控制酶解条件,蚯蚓可以利用自身的酶源自溶来代替外源酶,从而降低成本。     

鲟鱼鱼肠抗氧化肽的提取及抗氧化性

探讨酶法制备具有抗氧化活性的鲟鱼鱼肠抗氧化肽的方法,并进行体外抗氧化活性的测定。结果表明,比较4种蛋白酶酶解产物的抗氧化能力,确定胃蛋白酶为制备鲟鱼鱼肠抗氧化肽的最佳水解用酶;通过单因素试验和正交实验分析得出最适酶解工艺是:胃蛋白酶加酶量3 200 U/g,酶解时间1.5 h,料液比1∶20,温度35℃。其体外抗氧化能力随肽质量浓度增大而增大,在浓度为1.5 mg/mL时,鲟鱼鱼肠抗氧化肽清除DPPH·能力达到Vc的83.64%,对·OH清除率为78.06%,还原力大小约为Vc溶液的1/3。胃蛋白酶酶解鲟鱼鱼肠制备的抗氧化肽具有较好的抗氧化活性,其作为一种潜在的商业抗氧化剂具有良好的应用前景。     

胰蛋白酶酶解文蛤的工艺条件

用正交试验法确定胰蛋白酶酶解文蛤的最佳工艺条件 ,用薄层层析和氨基酸自动测定仪鉴定酶解液中氨基酸的种类和含量 ,进一步判定酶解效果。结果表明 :在 5 0℃ ,加酶量 2 %,pH8.0 ,酶解 8h的酶解效果最好。层析发现 ,酶解温度为5 0℃的提取液含氨基酸较全面 ,氨基酸自动测定仪测定显示 ,胰蛋白酶酶解法的文蛤提取液中氨基酸的含量大于热水提取法的含量。     

乳清分离蛋白酶解物的抗氧化活性研究

研究了乳清分离蛋白(WPI)酶解物对DPPH.、超氧阴离子自由基和羟基自由基的清除效果,同时用还原法研究了其抗氧化活性。结果表明,WPI酶解物在体外具有较强的抗氧化能力。木瓜蛋白酶酶解物和胰蛋白酶酶解物对DPPH.、超氧阴离子自由基、羟基自由基的清除能力和还原能力强于胰凝乳蛋白酶酶解物和胃蛋白酶酶解物。     

超高压酶解酪蛋白及其产物在动物细胞培养中的应用

以牦牛酪蛋白为底物,游离氨基酸含量为指标,在超高压条件下分别采用胰蛋白酶、胃蛋白酶、木瓜蛋白酶、中性蛋白酶、碱性蛋白酶和胰酶六种不同的酶水解酪蛋白并筛选出最佳用酶,并在单因素实验的基础上采用正交实验确定最佳用酶水解酪蛋白的最佳工艺条件。最后探讨水解产物在细胞培养中的应用。结果表明,胰酶水解效果最好,用其在底物浓度15%,E/S=1∶5、水解压力150 MPa,温度为45℃,pH 7.5条件下水解4h后水解产物中游离氨基酸含量高达52.18%。酶解产物对BHK-21细胞没有细胞毒性,而具有促生长作用。这为酪蛋白的进一步开发利用和蛋白质水解产物在细胞培养中的应用奠定基础。     

鲜鹿茸酶降解过程的研究

研究了胰蛋白酶、Alcalase 碱性蛋白酶、木瓜蛋白酶对鲜鹿茸的降解过程,确定了优化降解工艺条件,具有一定的理论意义和实践价值。确定了Alcalase 碱性蛋白酶的降解效率最高,通过单因素实验确定了降解过程中底物浓度、酶解温度、pH值和酶解时间为影响鲜鹿茸降解率的主要因素。正交试验确定最佳的酶解条件为:底物浓度0.08 g/ml、酶解温度65 ℃、pH 9.0、酶解时间6.0 h。在此条件下,鲜鹿茸降解率高达92.6%,氨基酸产品收率达12.1%。     

Relation between substrate feeding pattern and development of filamentous bacteria in activated sludge processes: Part III. applications with industrial waste waters

Summary Laboratory scale activated sludge systems were operated under regimes of continuous or intermittent feeding of the waste water. Industrial waste waters from breweries, a dairy plant and a petro-chemical plant were investigated. The systems were started up with sludge from a municipal waste water plant or more often with sludges obtained from the corresponding industrial waste water treatment plants. It was found that intermittently fed systems produced sludges with better settleability characteristics than systems that were continuously supplemented with waste water. Our previous hypothesis that in intermittent systems floc forming bacteria become dominant as a result of higher substrate uptake rates was confirmed and may thus be extended to waste waters containing readily available substrates such as carbohydrates (brewery-and dairy waste water) or acids (petro-chemical waste water). Supplementation of brewery waste water with urea had a negative influence on sludge settleability, especially in continuously operated systems.     

Pepsin as a catalyst for peptide synthesis: formation of peptide bonds not typical for pepsin substrate specificity.

Porcine pepsin in water solutions containing 15-28% of dimethylformamide at pH 5 and 20-37 degrees C catalysed the formation of peptide bonds between Z-Ala-Ala-Phe-OH and various amino acid or peptide derivatives. Substrate binding subsite S1' of pepsin demonstrated broad specificity in these reactions but revealed a certain preference for hydrophobic amino acid residues, including non-proteinous homophenylalanine, p-nitrophenylalanine, S-methylcysteine, as well as for those that contained, in addition to the hydrophobic elements, a group capable of donating a hydrogen bond, e.g. o-nitrotyrosine. This observation increases the range of peptides that might be prepared by pepsin-catalysed synthesis.     

Wound Healing Properties of Collagen from the Bone of Two Marine Fishes

Type I collagen was extracted from the bone of two marine fishes; Magalaspis cordyla and Otolithes ruber and its efficacy in wound healing was studied. The yields of acid soluble (30.5 and 27.6%) and pepsin soluble (45.1 and 48.6%) collagen were respectively as per their dry weight basis. Extracted protein was characterized as type I collagen basing on SDS PAGE, UV?CVis, FTIR spectrometer and amino acid composition. The characterized collagen was cross-linked with glutaraldehyde and found to possess three dimensional pores and acted efficiently in healing the excision wounds made on Wistar rats. Bone is generally considered as waste and discarded in many processing plant. But, the current study proved that it has excellent medicinal importance and reveals a possible way to convert this waste into a medicinally important source.     

氨基酸废水处理技术的研究进展

氨基酸废水处理技术一直是国内外氨基酸生产厂家和环保界的研究热点。本文对国内外氨基酸废水处理方法,如膜分离法、生物法等进行了详细评述,认为生物法是解决我国氨基酸废水污染的最终出路,同时提出将三维电极电化学法应用于氨基酸废水的处理,具有重要的理论意义和实际的应用价值。     

THE PROTEIN COMPOSITION OF NUCLEAR CRYSTALS IN LEAF CELLS

The chemical composition of perinuclear crystals of Lychnis chalcedonica and intranuclear crystals of Dianthus barbatus in leaf tissue embedded in glycol methacrylate was tested by differential enzyme digestion of ultra-thin sections. The Lychnis crystals were digested readily by pepsin but with great difficulty and never to completion by trypsin. The Dianthus crystals were rapidly and completely digested by pronase; inconsistently and with difficulty by pepsin; and were unaffected by trypsin. Pre-incubation with water increased the effectiveness of pepsin digestion. Neither RNase nor DNase had any effect, either alone, or when preceded or followed by the proteinases. It was concluded that the Lychnis crystals are composed of protein that is probably high in aromatic amino acid content, and possibly low in the basic amino acids arginine and lysine. The Dianthus crystals also seem to be protein, but because of their amino acid composition or conformation, and possibly because of complex reactions with the fixative, these crystals are not readily hydrolyzed by pepsin and trypsin.     

The purification and properties of a single chicken pepsinogen fraction and the pepsin derived from it

1. Evidence is given for the presence of at least five pepsinogens in a crude extract of mixed chicken stomachs. One of these was purified and could be activated to yield a single pepsin. 2. The molecular weights of the pepsinogen and pepsin were 36000 and 34000 respectively. The pepsin associated at low pH values and low ionic strength. 3. The amino acid analyses of both proteins are given. The pepsin was devoid of phosphate but contained carbohydrate. 4. The N-terminal amino acids of pepsinogen and pepsin were serine and threonine respectively. Five amino acids were released by carboxypeptidase A and it was deduced that serine may be the C-terminal one. 5. Each protein contained one thiol group per molecule as determined by titration with p-chloromercuribenzoate. The rate of the reaction was very rapid with pepsin, but much slower with pepsinogen, although the same group appeared to react in both instances. The enzymic activity of pepsin was unaffected by the modification. 6. The isoionic point of the pepsin was close to pH4.0 and the enzyme was stable for long periods at pH values up to 7.0. 7. The enzyme hydrolysed bisphenyl sulphite almost as rapidly as did pig pepsin A.     

Pepsinogen C and pepsin C: Further purification and amino acid composition

Pepsinogen C and pepsin C from the pig have been further purified by chromatography on DEAE-cellulose and by exclusion chromatography and the specific activities (with haemoglobin substrate) found are higher than those previously reported. The final preparations are homogeneous on electrophoresis in starch gel at three pH values except for contamination with less than 4% of pepsinogen and pepsin respectively. Pepsinogen C, like pepsin C, contains no phosphate. The amino acid compositions show some marked differences from those of pepsinogen and pepsin especially in the content of basic amino acids, glutamic acid, aspartic acid, leucine and isoleucine. The molecular weights of the enzyme and zymogen, obtained from the amino acid compositions, are 41400 and 36000 respectively, similar to those of pepsinogen and pepsin.     

Hydrolysis technology of biomass waste to produce amino acids in sub-critical water

Hydrolysis of biomass waste (such as fish waste, chicken waste, hair and feather) to produce amino acids was studied in sub-critical water, with reaction temperatures from 180 to 320 degrees C and reaction pressures from 3 to 30 MPa. The product of amino acid was determined by Amino Acid Analyzer (BioLC), and 18 kinds of amino acid were obtained. The results show that the controlling of reaction atmosphere, pressure, temperature and time of hydrolysis is very important to obtain high yield of amino acid; most of amino acids reached maximum yield at reaction temperature range of 200-290 degrees C and reaction time range of 5-20 min. There are obvious changes of amino acids yield at reaction pressures of 6-16 MPa and reaction temperature around 260 degrees C, owing to the homogeneousness of the first two phases of water in the formation of vapor and liquid. There are different yields of the same amino acid in different reaction atmospheres (e.g. air, carbon dioxide and nitrogen).     

Purification of coal gasification combined with brewery waste waters in upflow anaerobic sludge blanket digesters

Laboratory-scale upflow anaerobic sludge blanket (UASB) digesters containing granular sludge adapted to brewery effluent were fed increasing concentrations of artificial coal gasification (CG) waste water in brewery effluent and were effective at purifying this combined waste water up to a CG effluent concentration of 15%. At higher CG waste water concentrations and flow rates, performance declined rapidly.     

Fermentation of de-oiled algal biomass by <Emphasis Type="Italic">Lactobacillus casei</Emphasis> for production of lactic acid

De-oiled algal biomass (algal cake) generated as waste byproduct during algal biodiesel production is a promising fermentable substrate for co-production of value-added chemicals in biorefinery systems. We explored the ability of Lactobacillus casei 12A to ferment algal cake for co-production of lactic acid. Carbohydrate and amino acid availability were determined to be limiting nutritional requirements for growth and lactic acid production by L. casei. These nutritional requirements were effectively addressed through enzymatic hydrolysis of the algal cake material using α-amylase, cellulase (endo-1,4-β-d-glucanase), and pepsin. Results confirm fermentation of algal cake for production of value-added chemicals is a promising avenue for increasing the overall cost competiveness of the algal biodiesel production process.