二次中和法制取菜籽复合氨基酸新工艺研究

为得到高质量的菜籽复合氨基酸,以脱皮菜籽粕为原料,研究了硫酸水解制备复合氨基酸的新工艺一二次中和法,解决了氨基酸脱色和副产品植酸的利用问题。水解的最佳工艺参数是3mol·L-1的H2SO4水解18h;ca(OH)2作中和剂,第一次中和和脱色的最佳pH为2-4,第二次中和的最佳pH为6.5-7.0,中和后的溶液经减压浓缩,喷雾干燥,制得菜籽复合氨基酸。复合氨基酸得率≥40％,纯度≥40％;副产品植酸钙得率10％左右,纯度40％-50％。     

从鸡毛中提取复合氨基酸的研究

运用正交实验研究了以鸡毛为原料硫酸水解生产复合氨基酸的水解条件。结果表明 :硫酸浓度为 8mol·L- 1 ,鸡毛质量 (g)与硫酸体积 (mL)之比为 1∶2 .5 ,水解时间 10h ,其水解率可达 5 5 .2 3%。得到的固体氨基酸质量分数为 95 .31%。     

鸭毛梗制备复合氨基酸工艺条件的研究

采用正交试验方法 ,研究了鸭毛梗水解制备复合氨基酸的工艺条件 ,结果表明 :温度 /压力是影响氨基酸转化率最主要的因素。鸭毛梗水解制备的复合氨基酸转化率盐酸法最高 (82 .36 % ) ,氢氧化钠法最低 (5 8.4 6 % ) ,硫酸法与盐酸法接近(79.4 4 % )。硫酸法产率较高 ,操作方便 ,环境污染和设备腐蚀均小 ,适合大量生产 ,确定为水解制备复合氨基酸介质 ,其最佳工艺条件为 :水解时间 8.0h ,硫酸浓度 3.0mol·L-1,水解温度 12 5℃。氨基酸分析表明 ,水解液中均含有 18种以上氨基酸。     

不同生态系统土壤氨基酸氮的组成及含量

采集于内蒙古白音锡牧场、陕西澄城、杨凌、宜川和太白山等地不同生态系统的 1 2个土样 ,用 6 mol/ L HCl水解 ,经 H型酸性阴离子交换树脂柱纯化后 ,用 Beckman 1 2 1 MB型氨基酸分析仪测定了 1 7种常见氨基酸。测定结果表明 ,不同生态系统土壤酸解氨基酸含量有很大差异 ,表现为草甸土壤 (氨基酸含量为 2 2 83.9μg N/ g) >森林土壤 ( 1 733.6μg N/g) >草原土壤 ( 85 6 .3μg N/ g) >农田土壤 (平均为 2 4 8.5± 37.8μg N/ g) ,并且氨基酸氮与土壤全氮有极显著的正相关关系 ( p<0 .0 1 ) ;在氨基酸中以中性氨基酸所占比例最大 ,平均为 5 3.99% ,其次为碱性和酸性氨基酸 ,分别为 2 4 .94 %和2 0 .5 9% ,含硫氨基酸最少 ,仅为 0 .4 8% .游离氨基酸以草甸土壤最高 ,为 1 4 .5 8μg N/ g,其它土壤在 1 .1 4～ 8.6 7μg N/ g之间 ,大部分在 2～ 3μg N/ g。游离氨基酸不仅数量低 ,而且种类也比酸解氨基酸少。不管是酸解氨基酸 ,还是游离氨基酸 ,在 0～ 2 0 cm土层的含量均大于 2 0～ 4 0 cm土层 ,从不同土壤样品的平均结果看 ,对酸解氨基酸 ,0～ 2 0 cm土层为96 0 .9μg N/ g,2 0～ 4 0 cm土层为 5 2 8.9μg N/ g ;对游离氨基酸氮 ,0～ 2 0 cm土层 6 .2 8μg N / g,2 0～ 4 0 cm土层 2 .2 2μgN/ g。施用氮     

HD-I树脂在复合氨基酸脱色中的应用研究

HD-Ⅰ脱色树脂是一种大孔酚醛型弱酸树脂,对一些生化、有机产品具有较强的脱色能力,其优点为脱色时树脂用量小、易再生、经再生后可以反复使用。该树脂在复合氨基酸的脱色中效果优于颗粒状活性炭。带有酱油色、透光率为20%左右,5%的复合氨基酸经HD-Ⅰ树脂脱色后其外观为无色、清亮、透明的溶液,透光率可达97%左右,同时氨基酸的收率在95%以上。     

一株产环糊精葡萄糖基转移酶的地衣芽孢杆菌的选育、产酶条件及酶学特性

从土壤分离物中筛选到一株环糊精葡萄糖基转移酶 (CGTase)产生菌 4 0 3,96h发酵酶活为 0 95U mL。经紫外辐射和硫酸二乙酯复合诱变而获得突变株CLS4 0 3,96h发酵酶活达 1 36U mL ,提高 4 3%。该突变菌株被鉴定为地衣芽孢杆菌 (Bacilluslicheniformis) ,产CGTase的最佳碳源为可溶性淀粉 ,最佳氮源为硝酸铵 ,最适初始pH为 6 5 ,最适培养温度为 35℃ ,发酵期间CGTase的产生高峰 (第 96h)滞后于菌体生物量高峰 (第 4 8h) 2d。菌株所产CGTase的最适反应pH为 6 0 ,最适温度为 5 5℃ ,在pH 6 0～ 7 5间和 5 0℃下保持 1h后的剩余酶活均达 90 %以上 ;酶液中适量添加Ca2 能大幅提高CGTase在 5 5℃下的稳定性。经高效液相色谱分析 ,CGTase作用于淀粉后的产物以α 环糊精为主 ,β 环糊精为次 ,二者比例为 2 4 7∶1,环糊精总产率达 2 9 8% ,但产物中不含γ 环糊精     

不同品种魔芋精粉中氨基酸含量分析研究

花魔芋和白魔芋精粉中均含有 18种水解氨基酸和 18种游离氨基酸 ,前者水解氨基酸的总量为 2 .4 4 % ,其中必须氨基酸质量分数为 0 .6 2 % ,游离氨基酸总量为 1.10 % ,其中游离必须氨基酸质量分数为 0 .33% ;后者水解氨基酸总量为 2 .0 1% ,其中必须水解氨基酸质量分数为 0 .5 6 % ,游离氨基酸总量为 0 .4 1% ,其中必须游离氨基酸质量分数为 0 .13%。提示白魔芋和花魔芋精粉中氨基酸含量略有差异 ,但此差异不大 ,其品质 ,营养价值及特性等主要取决于魔芋精粉中主要成分葡甘聚糖的含量     

克氏螯虾虾壳制备氨基葡萄糖盐酸盐的工艺研究

研究了以克氏螯虾壳为原料制备氨基葡萄糖盐酸盐 (GAH)的工艺条件 :酸水解液为 10mol·L-1HCl,85～ 92℃水解 5～ 6h ;甲壳素∶酸水解液为 3∶5。GAH产率为甲壳素的 5 2 %～ 5 5 %。GAH纯度 :99%～ 10 0 .5 %。     

稀有鮈鲫的脂肪酸组成

稀有鲫汉源种群和彭州种群全鱼中共检测到 3种饱和脂肪酸 (SFA) ,它们是C1 4∶0、C1 6∶0、C1 8∶0。 4种单不饱和脂肪酸 (MUFA) ,它们是C1 4∶1、C1 6∶1、C1 8∶1、C2 0∶1。 4种多不饱和脂肪酸 (PUFA) ,它们是C1 8∶2、C2 0∶4、C2 0∶5 (EPA)、C2 2∶6 (DHA)。SFA占总脂肪酸的2 3 6 3— 2 8 97% ,MUFA占 40 73— 5 4 3 2 % ,PUFA占 9 96— 2 3 1 7% ,EPA占 0 41— 1 74% ,DHA占 0 1 1— 5 3 7%。EPA含量一般低于DHA。UFA(MUFA +PUFA) % >SFA % ,且MUFA % >PUFA % ,n 3FA/n 6FA为 0 3 3— 0 99。此外还测定了鱼肌、内脏脂肪、肝脏、性腺等的脂肪酸组成和含量     

Xterra RP18柱高效液相色谱法快速分离测定氨基酸

建立了一种用XterraRP1 8色谱柱快速分离测定水解氨基酸的方法。所采用的色谱条件是 :WatersAlliance系统 ,柱温 5 6℃ ,流速 1 .8ml/min ,检测波长 2 4 8nm ,梯度分离 ,运行周期 2 5min,柱反压低于 2 0 0 0Psi。在 1 7.5min内分离了包括AMQ、NH3 和牛磺酸在内的 2 1种氨基化合物 ,适应于复合氨基酸注射液、含牛磺酸的氨基酸口服液及水解氨基酸样品的分析测定     

Electrostatic control by lipids upon the membrane-bound (Na+ + K+)-ATPase. II. The influence of surface potential upon the activating ion equilibria

Sarcoplasmic reticulum Ca2+-ATPase has been reconstituted with excess lipid (25-150 g egg phosphatidylcholine per g sarcoplasmic reticulum protein) by a procedure combining the use of a non-ionic detergent with cholate dialysis. The reconstituted vesicles were analyzed by sucrose density fractionation and freeze-fracture electron microscopy. At the lowest lipid to protein ratios some vesicles containing aggregated protein were observed. At a lipid to protein ratio of 150:1 (w/w) only 30-40% of the reconstituted protein sedimented through 7% (w/v) sucrose. The remainder of the latter preparation was characterized by a high Ca2+-uptake capacity and a coupling ratio of 1.6 mol Ca2+ transported per mol ATP hydrolyzed. Intramembranous particles in this preparation occurred isolated in the membrane. In most cases only one particle could be seen on a fracture face. Cross-linking with cupric phenanthroline indicated that protein-protein contacts were drastically reduced by reconstitution. It is concluded that aggregation of intramembranous particles is not required for optimal Ca2+-transport function. The dispersed preparation obtained by a combined reconstitution and sucrose density fractionation procedure is useful for further characterization of the Ca2+ pump.     

重组萝卜磷脂氢谷胱甘肽过氧化物酶在毕赤酵母优化表达初步纯化与鉴定

将萝卜磷脂氢谷胱甘肽过氧化物酶（RsPHGPx）基因插入到分泌表达载体pPIC9K中,转化巴斯德毕赤酵母GS115细胞,筛选具有G418抗性的单拷贝转化子。经过优化表达条件,RsPHGPx在1%甲醇、pH6.0、28℃条件下诱导60h后得到最大表达量,产率约为102 mg/L。通过硫酸铵分级沉淀、脱盐柱脱盐、凝胶过滤等纯化步骤,得到了90%以上纯度的RsPHGPx．活性分析显示纯化获得的RsPHGPx具有依赖于GSH的还原活性, 比活性为4.2μmol/min·mg,为获得大量RsPHGPx而用于应用开发研究奠定了基础。     

Dicyclohexylcarbodiimide-sensitive ATPase in Halobacterium saccharovorum

Membranes from Halobacterium saccharovorum contained a cryptic ATPase which required Mg2+ or Mn2+ and was activated by Triton X-100. The optimal pH for ATP hydrolysis was 9-10. ATP or GTP were hydrolyzed at the same rate while ITP, CTP, and UTP were hydrolyzed at about half that rate. The products of ATP hydrolysis were ADP and phosphate. The ATPase required high concentrations (3.5 M) of NaCl for maximum activity. ADP was a competitive inhibitor of the activity, with an apparent Ki of 50 microM. Dicyclohexylcarbodiimide (DCCD) inhibited ATP hydrolysis. The inhibition was marginal at the optimum pH of the enzyme. When the ATPase was preincubated with DCCD at varying pH values, but assayed at the optimal pH for activity, DCCD inhibition was observed to increase with increasing acidity of the preincubation medium. DCCD inhibition was also dependent on time of preincubation, and protein and DCCD concentrations. When preincubated at pH 6.0 for 4 h at a protein:DCCD ratio of 40 (w/w), ATPase activity was inhibited 90%.     

高温蛋白酶制备酶解猪血蛋白的研究

用高温蛋白酶ＷＦ１４６制备猪血水解蛋白得率在１０％（Ｗ／Ｖ）以上。水解液经分析含有１８种氨基酸,其中８种必需氨基酸占３７．１４％。氨基酸总量可达６２６４．８６ｍｇ／１００ｍｌ．产物具有较高的营养价值和良好的溶解性,因而在食品工业中具有很大的应用潜力。     

Laboratory-scale inactivation of African swine fever virus and swine vesicular disease virus in pig slurry

Two methods were evaluated for the inactivation of African swine fever (ASV) and swine vesicular disease (SVD) viruses in pig slurry: chemical treatment and heat treatment. The addition of NaOH or Ca(OH)2 at different concentration/time combinations at 4 degrees C and 22 degrees C was examined, as was virus stability at different temperature/time combinations. ASF virus (ASFV) was less resistant to both methods than SVD virus (SVDV). In slurry from one source, ASFV was inactivated at 65 degrees C within 1 min, whereas SVDV required at least 2 min at 65 degrees C. However, it was found that thermal inactivation depended on the characteristics of the slurry used. Addition of 1% (w/v) of NaOH or Ca(OH)2 caused the inactivation of ASFV within 150 s at 4 degrees C; 0.5% (w/v) NaOH or Ca(OH)2 required 30 min for inactivation. NaOH or Ca(OH)2 (1% (w/v)) was not effective against SVDV at 22 degrees C after 30 min, and 1.5% (w/v) NaOH or Ca(OH)2 caused inactivation of SVDV at both 4 degrees C and 22 degrees C. At higher chemical concentrations or temperatures, ASFV and SVDV inactivation was faster in slurry than in buffered medium.     

单宁酶的固定化及其在酯型儿茶素水解反应中的应用

采用自制的壳聚糖为载体对单宁酶（ＴＡ）固定化,ＴＡ与壳聚糖配比１：２．５,３０℃固定２ｈ,活力回收达２３．６％～３３．１％;偶联效率为８４．９％～８８．０％。固定化单宁酶（ＩＴＡ）的表观Ｋｍ值（以没食子酸丙酯为底物）为２２．２×１０－６ｍｏｌ／Ｌ,ＴＡ的Ｋｍ值（以没食子酸丙酯为底物）为１０×１０－６ｍｏｌ／Ｌ,ＴＡ和ＩＴＡ的最适反应温度分别为４０℃和５０℃;６０℃处理１５ｍｉｎ,残存活性分别为１３．６％和６０．３％。ＴＡ和ＩＴＡ的最适ｐＨ值分别为５．８和６．４;ＴＡ在ｐＨ４．８～７．８活力稳定,而ＩＴＡ活力稳定范围在ｐＨ４．８～６．８．ＩＴＡ作用于ＥＧＣＧ的半衰期为７８．７ｈ,ＥＧＣＧ水解率达９０．３％。对茶多酚提取物进行水解,其所含的酯型儿茶素ＥＧＣＧ和ＥＣＧ水解率分别为９６．４％和９６．８％,非酯型儿茶素ＥＧＣ和ＥＣ的含量显著增加。     

Optimization of butanediol dimethacrylate crosslinked polystyrene: A novel polymeric support for solid phase peptide synthesis

Summary Studies leading to optimization of butanediol dimethacrylate-crosslinked polystyrene supports (BDDMA-PS) for solid phase peptide synthesis are delineated. BDDMA-PS copolymers with different crosslink densities were prepared and functionalised with chloromethyl groups. The reactivity of the Lys(2-Cl−Z)−OH residue bound to these polymers through a benzyl ester linkage was investigated by following the kinetics of acylation by the HOBt active ester of Boc-Alanine. From the results it was observed that the rate of peptide bond formation was maximum for a 2% BDDMA crosslinked resin. This resin was compared with a 2% DVB-crosslinked polystyrene resin (DVB-PS). Synthesis of an extremely insoluble, hydrophobic, antiparallel β-sheeted difficult sequence peptide LMVGGVVIA (β 34–42), C-terminal fragment of β-amyloid protein, β (1–42), was carried out on both 2% DVB-PS and 2% BDDMA-crosslinked polystyrene supports. The synthesis of the peptide was carried out using Boc amino acid strategy. Greater extent of swelling of the resino peptide, increased coupling efficiency during the assembly of amino acids and relatively high purity of synthesised peptide were observed in the case of 2% BDDMA-PS polymer.     

Protein Metabolism in Cultured Plant Tissues: III. Changes in the Rate of Protein Synthesis, Accumulation, and Degradation in Cultured Pith Tissue

During the transition of tobacco (Nicotiana tabacum) pith tissue to callus tissue, there were changes in the composition of the soluble amino acid pools, in the distribution of amino acids between pool and protein, and in the synthesis, accumulation, and degradation of proteins. The size of the leucine pool decreased from 90 nanomoles per gram fresh weight in fresh pith to 20 nanomoles in 24-hour cultured pith, followed by a return to 90 nmoles in pith cultured longer than 5 days. The latter value is the same as that reported for exponentially growing callus cells. Many other pool amino acids changed as dramatically. However, they always approached callus levels after 5 days of culturing. The total amino acid content of pith tissue (the sum of both pool and protein) remained unchanged during culturing. The value for total amino acid content (34 to 42 nanomoles per gram fresh weight) was also similar to that found in callus. The distribution of amino acids between pool and protein did change during culturing. The transition of pith tissue with 88% of its total amino acids free in the soluble pool to callus with 92% of its amino acids in protein was further characterized by changes in protein metabolism. Both protein synthesis and accumulation increased over the first 50 hours in culture to a maximum rate of 45 milligrams protein synthesized gram protein⁻¹ hour⁻¹. After 50 hours in culture, the rate of protein accumulation decreased to equal the rate of fresh weight accumulation (10 mg g⁻¹ hour⁻¹). However, protein synthesis continued at a high rate for several days, suggesting protein degradation was turned on by this time. By 5 days protein synthesis had decreased to a rate similar to that of callus.     

Purification and properties of extracellular matrix-degrading metallo-proteinase overproduced by Rous sarcoma virus-transformed rat liver cell line, and its identification as transin

Rous sarcoma virus-transformed rat liver cell line RSV-BRL secreted a neutral proteinase in a latent precursor form with a molecular weight (Mr) of 57,000 (57k) as a major secreted protein. This enzyme was a calcium-dependent metallo-proteinase. The proenzyme was purified from the serum-free conditioned medium of the transformed cells by affinity chromatographies on a zinc chelate Sepharose column and a reactive red agarose column. When activated by treatment with trypsin or p-aminophenylmercuric acetate (APMA) in the presence of Ca2+, the purified enzyme effectively hydrolyzed casein, fibronectin, and laminin. Type IV collagen was hydrolyzed at 37 degrees C but not at 30 degrees C by the enzyme, whereas type I and type III collagens were hardly hydrolyzed even at 37 degrees C. The treatment with trypsin or AMPA in the presence of Ca2+ converted this 57k proenzyme to an active and stable enzyme with Mr 42k. In the absence of Ca2+, however, APMA converted the proenzyme to an intermediate form with Mr 45k, while trypsin digested it to an inactive peptide with Mr 30k. These results demonstrate that calcium ion is essential for the activation, activity expression, and stabilization of this metallo-proteinase. Analysis of its partial amino acid sequence and amino acid composition showed that the 57k proenzyme was identical or closely related to the putative protein transin, a rat homologue of stromelysin.