Flavourzyme-Papain复合蛋白酶水解龙虾副产物工艺条件的研究

以底物浓度为10%、Flavourzyme与Papain复合酶比为1:1的条件,利用单因素试验和正交试验,研究了pH值、加酶量、温度、酶解时间对龙虾副产物蛋白质水解度的影响,在此基础上进一步确定了合适的底物浓度。结果表明,Flavourzyme-Papain复合蛋白酶水解龙虾副产物的最佳条件为:pH7.0、加酶量1.5%、温度50～55℃、时间5h,在底物浓度25%的条件下水解,最大水解度为26.2%。

CNBr-Papain-Sepharose 4B亲和填料的制备及其性质研究

为探求从鱼类废弃组织及漂洗水中回收Cystatin的高效方法,首先自制了溴化氢-木瓜蛋白酶-琼脂糖凝胶(CNBr-Papain-Speharose 4B)亲和填料,确定了适宜的配基Papain偶联密度,之后测定了该亲和填料的热稳定性、酸碱稳定性、贮藏性能及使用的重复性,最后以该制备的填料对鱼糜漂洗水中的CPIs活性进行了初步回收。以对重组鲢鱼Cystatin吸附率为评价指标,确定适宜Papain偶联密度,即CNBr-Speharose 4B和Papain溶液最佳料液比为1∶7(g∶mL)。CNBr-Papain-Speharose 4B的适宜使用温度为4~40℃,且在pH 3.0~12.0之间稳定。CNBr-Papain-Speharose 4B在4℃下,6个月的贮藏期内,表现较好的稳定性。将填料装柱后进行验证实验,证明该填料的使用重复性能稳定,且最终成功高效地回收了鲢鱼鱼糜漂洗水中的CPIs。

An Orthogonal Experiment of Papain-Assisted Extraction of Polysaccharides from Agrocybe aegirit

[Objective] This study aimed to explore optimum conditions for papain-as- sisted extraction of polysaccharides from Agrocybe aegirit. [Method] Because enzyme is capable of destroying organizational structure of cells, papain was selected to cat- alyze cells to let polysaccharides out. An orthogonal experiment was carried out to investigate the effects of hydrolysis temperature, hydrolysis time, enzyme concentra- tion and solid to liquid ratio on extraction of polysaccharides from A. aegirit. [Result] Through the orthogonal experiment, the optimum papain-assisted extraction conditions were obtained： hydrolysis temperature, 45 ℃; hydrolysis time, 2 h; enzyme concen- tration 2%; solid to liquid ratio, 1：80. [Conclusion] This study will provide a reference for extraction of polysaccharides from A. aegirit.

Inhibition of cysteine protease papain by metal ions and polysulfide complexes,especially mercuric ion

Aim Cysteine proteases are closely associated with many human and non-human pathological processes and are potential targets for metal ions especially Hg^2＋ and the related species. In the present work, on the basis of to the general study on the effects of some metal ions on the activity of papain, a well-known representative of cysteine protease family, the inhibitory effects of Hg^2＋ and polysulfide complexes were studied. Results All the metal ions tested （Hg^2＋, Cu^2＋, Ag^＋, Au^3＋, Zn^2＋, Cd^2＋, Fe^3＋, Mn^2＋, Pb^2＋, Yb^3＋） inhibit the activity of papain anda good correlation between the inhibitory potency and softness-and-hardness was observed. Among the metals, Hg^2＋ was shown to be a potent inhibitor of papain with a Kiof 2 × 10^-7 mol·L^-1 among. Excessive amounts of glutathione and cysteine could reactivate the enzyme activity of papain deactivated by Hg^2＋. These evidences supported that Hg^2＋ might bind to the catalytic site of papain. Interestingly, Hg （Ⅱ） polysulfide complexes were for the first time found to inhibit papain with a Ki of 7 × 10^-6 mol·L^-1, whose potency is close to a well known mercury compound, thimerosal （Ki=2.7 × 10^-6）. In addition, Hg （Ⅱ） polysulfide complexes exhibit good permeability （ 1.9 × 10-5 cm· s^-1） to caco-2 monolayer. Conclusion These results suggested that mercury polysulfide complexes might be potential bioactive species in the interaction with cysteine proteases and other- SH-content proteins, providing a new clue to understand the mechanism of the toxicological and pharmacological actions of cinnabar and other insoluble mercury compounds.

Effects of Papain Hydrolysis on the Pasting Properties of Wheat Flour

As one of the most effective enzymatic modification methods of protein, papain hydrolysis is applied widely in food production, accompanying starch pasting frequently in order to improve industrial quality. Effects of the papain hydrolysis on flour pasting properties were investigated in five papain/flour concentrations and five time-treatments. The structure of starch and protein networks in slurry was investigated under microscope before and after pasting. Results showed that papain hydrolysis influenced the pasting properties of wheat flour significantly through affecting structural characteristics, amylase activity and exotbermic transition, especially during the early stage of hydrolysis. Peak viscosity, trough, final, integral area, and setback significantly decreased along with the increasing concentration of papain. Both hydrolysis time and concentration of papain had obviously effect on the breakdown. Pasting temperature and pasting time increased significantly with the enhancement of papain concentration. Hydrolysis time exerted minor effect on the pasting temperature and pasting time. The average peak time was slightly prolonged by lower concentration of papain, otherwise slightly shortened by higher concentration.

Immobilization of Papain in Biosflica Matrix and Its Catalytic Property

A novel method was developed for papain immobilization through a biomimetic silicification process induced by papain. By incubating papain in a silica precursor solution, the papain-silica composite formed rapidly and oanain was encansulated. The encansulation efficiency and the recovery activity were 82.60% and 83.09%, re-spectively. Compared with enzymes and biomolecules immobilized in biosilica matrix in the presence of additaonal silica-precipitating species, this papaln encapsulation process, a biomimetic approach, realized high encapsulation efficiency by its autosilification activity under mild conditions （near-neutral pH and ambient temperature）. Fur-thermore, the encapsulated papain exhibits enhanced thermal, pH, recycling and storage stabilities. Kinetic analysis showed that the biomimetic silica matrix did not significantly hinder the mass transport of substrate or the release of product.

Ag-induced Efficient Immobilization of Papain on Silica Spheres

Complexation and interaction between silver and amino group were applied to induce an efficient immobilization of papain on silica spheres.Tbe silver nanoparticles were deposited on the silica spheres before p apainwas coupled to the silica spheres. The silica spheres with silver nanoparticles were characterized by high resolution transmission electron microscopy （HR-TEM）, Fournier transform infrared spectroscopy （FT-IR）, and UV-Vis scanning spectrometer. FT-IR spectrum was also used to characterize the immobilized and free papain. Effect of some factors on the activities of the immobilized papain was investigated. It was observed that the coupled yield and relative activity of the papain on Ag/SiO2 were 1.17 and 1.86 times of those on the bare SiO2, respectively. At an optimum concentration of silver, theobserved activity of the immobilized papain was 2.1 timesof that on the bare.silica.In addition, the maximum specific activity of papain immobilized on Ag/SiO2 was 819.9 U·mg·^-1, which is slightly lower than that of the free papain, 906.2 U·mg^-1 . Stability of the immobilized papain was also examined. The resuits indicate that the silver nanoparticles successfully induce a fine immobilization of papain.

黑芥和埃塞俄比亚芥PAP2基因克隆及B基因组PAP2功能验证

PAP2(Production of anthocyanin pigment 2)编码MYB转录因子,与bHLH、WD40形成MYB-bHLH-WD40复合体来调控花青素合成。本研究从埃塞俄比亚芥和黑芥中分别克隆出2个和1个PAP2基因拷贝,分别命名为BcaB.PAP2、BcaC.PAP2、BniB.PAP2,这三个基因均由3个外显子和2个内含子组成,均编码247个氨基酸。基于本研究克隆的PAP2基因序列和已报道的其他芸薹属PAP2基因序列,设计了3对能分别检测A、B、C基因组PAP2基因的引物,通过等位特异PCR可以区分芸薹属禹氏三角中的6个物种PAP2的基因组来源。选取芥菜型油菜B基因组的BjuB.PAP2基因构建过表达载体,转化拟南芥,其叶片变紫,表明PAP2基因正调控花青素合成。遮光处理10天后紫叶埃塞俄比亚芥的叶片紫色变淡,花青素含量下降了41.22%。荧光定量PCR和转录组研究表明,遮光处理植株叶片中BcaB.PAP2和BcaC.PAP2表达量均下降,花青素合成的结构基因查尔酮合成酶基因(CHS)、查尔酮异构酶基因(CHI)、黄烷酮3-羟化酶基因(F3H)、二氢黄酮醇还原酶基因(DFR)、花青素合成酶基因(ANS)、类黄酮3-葡糖基转移酶基因(UFGT)等基因的表达量也下降。本研究完成了对芸薹属植物中埃塞俄比亚芥和黑芥PAP2基因的克隆,并对其进行进化分析。根据本实验中克隆及其他已报道的PAP2基因序列设计等位基因特异性PCR引物,为芸薹属种间杂交后代PAP2基因的基因组传递鉴定提供了分子标记。通过BjuB.PAP2基因转拟南芥植株结果表明PAP2基因参与花青素的积累。紫叶埃塞俄比亚芥经遮光处理后,发现BcaPAP2基因及花青素合成结构基因的表达受光照的诱导。本研究在芸薹属中克隆PAP2基因,并初步验证PAP2的功能,为进一步解析芸薹属植物花青素合成的调控机制提供参考。

THERMAL ACTIVATION OF IMMOBILIZED PAPAIN

Papain (Papainase, EC 3.4.22.2) was immobilized on porous silica beads by cross linking with glutaraldehyde. The thermal activation of this immobilized papain in aqueous system was found at, a temperature range from 50 to 90 degrees C. The higher the temperature, the more active the immobilized papain will possess. At the same time, the durability of the immobilized papain on heating was greatly improved. The effect of additives and salts on the activity of the immobilized papain were also studied. The results showed that the additives and some of the salts studied could markedly enhance the activity of the immobilized papain at elevated temperature.

STUDY ON THE IMMOBILIZATION OF PAPAIN WITH A MACROPOROUS BEAD CARRIER OF COPOLYMER CONTAINING MONOMER UNITS OF NAMINOETHYL ACRYLAMIDE AND VINYL ALCOHOI

A kind of macroporous bead carrier of copolymer containing monomer units of N-aminoethyl acrylamide and vinylalcohol was synthesized, i.e. the MR-AA carrier. Papain was immobilized on the carrier using glutaraldehyde as the couplingagent. The enzymatic activity of the immobilized papain was compared with free papain using casein as a substrate, and theeffects of glutaraldehyde concentration, pH, temperature, time and papain amount added on the activity recovery were alsoinvestigated. The results show that the MR-AA carrier contains reactive primary amine groups, hydrophilic amido links andhydroxyl groups, as well as macroporous structures based on its matrix (MR-AV matrix), furthermore, the activity recoveryof papain in the immobilization could reach 48%/～58%. In comparison with free papain, the resulting immobilized papainexhibits a remarkable thermostability and better reusability.

Optimization of Adsorption Conditions of Papain on Cross-Linked Phage-Displayed Heptapeptide Matrix Using Response Surface Methodology

A new cross-linked heptapeptide matrix(CHM)was prepared and the adsorption performance for papain was investigated.Firstly,a phage-displayed heptapeptide(Ile-Gln-SerPro-His-Phe-Phe)with high affinity to papain was found.Secondly,the heptapeptide was cross-linked to a matrix for papain adsorption.Finally,the adsorption conditions were optimized by response surface methodology.The results indicated that the optimum adsorptionconditionsweredeterminedasinitialpH 6.9,temperature 40℃,and the adsorption capacity of CHM for papain was found to be 55.52 mg/g.Moreover,the papain was purified 78-fold in a single step determined by affinity precipitation.More than87%of the adsorbed papain was desorbed using the eluent solventtriethylamine at pH 7.4.The results show that the CHM is a promising adsorbent for papain purification from crude papaya powder.

Papain Activity in Dextran Solution for Keratin Hydrolysis

High concentration of polymer solution in cosmetics has the essential role in keeping water on peel of skin. This study includes the papain activity in the high concentrated dextran solution for keratin hydrolysis. Papain is the industrial and a representative hydrolase. The concentration of dextran was ranged to 100 g/L. In the increasing concentration of dextran, the activity of papain decreased due to the low diffusivity of papain and substrate. Even in concentrated dextran solution, keratin powder was possible to be hydrolyzed at the high efficiency.

PAP-ZnCl_(2)浸渍增强木材的耐紫外及力学性能

为提高木材的耐紫外线和力学性能,采用聚氨基聚醚亚磷酸酯(PAP)和氯化锌(ZnCl_(2))复合浸渍剂对木材进行改性处理。通过浸渍处理,评估了浸渍条件下,PAP-ZnCl_(2)对木材性能的影响。结果表明,PAP-ZnCl_(2)浸渍处理显著改善了木材的紫外线耐受性和力学性能。具体而言,处理后的木材表现出优异的紫外线阻隔效果和较高的拉伸性能及弯曲性能,优于未处理的对照样本。改性后的木材拉伸性能由20.5 MPa提高至41.3 MPa,弯曲性能由80.3提高至94.25 MPa,并且改性后的木材在波长为200~600 nm处的紫外屏蔽效果显著。红外光谱(FTIR)、扫描电子显微镜(SEM)和力学性能测试表明,PAP和ZnCl_(2)的共同作用有效增强了木材的表面结构稳定性,减少了因紫外线照射引起的材料降解。研究结果为木材在户外环境中的耐久性提升及相关领域的实际应用具有参考价值。

Preparation of Dendrimers from Macromolecular Cores and Papain Immobilization

Using amino-poly(ethylene glycol) and amino-propyl silica gel as initiator cores, two dendrimers were prepared by a two-step procedure. The progress of dendrimers during each step was monitored by infrared spectroscopy and elemental analysis. Papain was crosslinked to the solid dendrimer with silica gel core by glutaraldehyde, and the enzymatic activity of the immobilized papain kept almost unchanged after repeated usage for fifteen times.

PAP效应下后蹲对田径运动员冲刺和垂直跳跃能力的影响

文章探讨了PAP效应下后蹲对田径运动员冲刺及垂直跳跃能力的影响。研究表明,恰当的后蹲训练可以通过神经肌肉的活化增强运动员的爆发力,从而提高运动员的冲刺速度及跳跃表现。文章对后蹲产生的机理及其促进冲刺技术与跳跃动作的效果进行了分析,建议制定综合性后蹲训练方案,内容涉及训练频率、强度及动作变化等方面,同时应注重对训练过程的监测和调节。文章对田径教练在训练过程中充分发挥PAP效应、优化训练成效等方面具有一定的实践指导作用。

不同形式诱导的PAP对短跑运动员下肢等长肌力的影响研究

该研究旨在探讨不同形式诱导的激活后增强效应(PAP)对短跑运动员下肢等长肌力的影响。通过实验法,以12名男性短跑运动员为实验对象,分别进行45%1RM、120 mmHg加压血流限制结合45%1RM、80%1RM的负重半蹲练习,收集各时间段内的测试结果。结果显示:在120 mmHg加压血流限制结合45%1RM负荷干预下,受试者下肢等长肌力(前群)成绩要优于45%1RM负荷下的成绩,在第7分钟成绩相差最大,P<0.05,具有显著性差异;与45%1RM相比,在80%1RM负荷下,受试者在第1分钟的成绩就有较高提高,P<0.01,具有非常显著性差异,之后在第7分钟成绩相差最大,P<0.01,具有非常显著性差异;120 mmHg加压血流限制结合45%1RM与80%1RM负荷干预相比,实验后各恢复时间点的等长肌力(前群)成绩均无显著性差异;与45%1RM相比,在120 mmHg加压血流限制结合45%1RM负荷干预下,受试者下肢等长肌力(后群)成绩在第7分钟至第16分钟要优于45%1RM等长肌力(后群),但整体P>0.05,不具有显著性差异;在80%1RM负荷干预下,受试者第10至13分钟成绩与45%1RM成绩相差最大,P<0.05,具有显著性差异;与120 mmHg加压血流限制结合45%1RM相比,在80%1RM负荷下,受试者下肢等长肌力(后群肌)成绩更优,但从实验后各恢复时间点来看,两种负荷下的成绩P>0.05,无显著性差异。该研究为短跑运动员的训练提供了理论依据,教练员可根据运动员的具体情况和训练目标,选择合适的负荷干预方式,以提高其运动表现。

“PAP”课堂教学模式在中职信息技术课程教学中的应用

信息技术课程作为中职学生的必修课程之一,在课程教学过程中,如何提高学生的学习积极性,满足学生的个性化学习需求以及实时反馈课堂教学质量和学习效果评价,是相关教师重点关注的课题。基于此,本文以中职信息技术课程为例,对“PAP”课堂教学模式的应用展开研究,首先阐述了“PAP”课堂教学模式,然后围绕中职信息技术课程教学中存在的问题进行分析,并提出相应的措施。最后,经实践证明“PAP”课堂教学模式能够完善信息技术课程的教学方式,对提高学生的学习技能和综合素养具有重要的应用价值。

PAPS免疫微球快速诊断伊氏锥虫病的研究

本文介绍一种新型的PAPS锥虫快速诊断试剂。PAPS是我组研制成功的一种新型的载体微球,它与伊氏锥虫抗原交联所制备的锥虫快速诊断液具有特异性强、敏感性高、重复性好、快速、简便的特点。我们应用锥虫快诊液检测212头锥虫阳性病牛(见虫),阳性检出率为98.6％。对275头非疫区阴性牛进行检测,阴性符合率为99.6％。PAPS试验与血凝试验的符合率为93.2％。对142头血清和血纸两种血样本进行PAPS试验的阳性结果和反应强度完全吻合,无明显差异。

GPC-3、PAP、EPCA-2及联合PSA检测在前列腺癌中的诊断价值

目的探讨血清磷脂酰肌醇蛋白聚糖-3(phosphatidylinositol proteoglycan-3,GPC-3)、前列腺酸性磷酸酶(prostatic acid phosphatase,PAP)、前列腺癌抗原-2(prostate cancer antigen-2,EPCA-2)及联合前列腺特异性抗原(prostatic specific antigen,PSA)检测在前列腺癌中的诊断价值.方法选取确诊的78例前列腺癌患者(前列腺癌组)和50例良性前列腺增生患者(前列腺增生组)作为研究对象,并选取同期与之配对的80例男性健康体检者作为对照组.用酶联免疫吸附试验检测血清GPC-3、PAP和EPCA-2水平,用时间分辨荧光分析法检测血清PSA水平,并绘制受试者工作特征(ROC)曲线分析GPC-3、PAP、EPCA-2和PSA对前列腺癌的诊断价值.结果前列腺癌组、前列腺增生组和对照组血清GPC-3、PAP、EPCA-2和PSA依次降低,3组比较差异均有统计学意义(P<0.05).血清GPC-3、PAP、EPCA-2和PSA 4个单项指标对前列腺癌均有一定的诊断价值,其敏感度及特异度分别为62.48%、63.31%,64.82%、60.19%,66.55%、63.33%,68.15%、65.92%(P=0.041、0.045、0.038、0.024);但联合检测可明显提高其诊断效能,以GPC-3+PAP+EPCA-2+PSA联合检测效果最佳,其敏感度和特异度分别为95.78%和92.89%.结论血清GPC-3、PAP、EPCA-2及PSA对前列腺癌均有一定诊断价值,但联合检测可明显提高其诊断效果,尤以GPC-3、PAP、EPCA-2及PSA四者联合效能最佳.