Effect of Weaning Ages on Pancreatic and Intestinal Chymotrypsin Activity in Piglet

Twelve litters of new born piglets were divided randomly into groups I , I , M and F , and weaned at 17, 21, 28 and 35 days of age, respectively, to determine the effect of weaning age on pancreatic and intestinal chymotrypsin activity in piglets. The results showed that the relative and specific activity of pancreatic chymotrypsin increased significantly 12h after weaning compared with that of sucking piglets at the same age and then decreased and remained low for 2 - 3 weeks after weaning. Total pancreatic chymotrypsin activity had no change with the increase of pancreas weight during day 18 - 50 regardless of weaning (P > 0.05). Chymotrypsin activity of jejunum had the same change as that of pancreas during sulking stage. Jejunum chymotrypin activity decreased in the first week post-weaning in the piglets weaned before day 28, but had no change in groups weaned at 28 or 35 days. The earlier the weaning age, the longer the restoring time. Chymotrypsin activity in jejunum is more sensitive to the effects of weaning age than in duodenum and ileum.

SNP Analysis and Haplotype Identification in Chymotrypsin Inhibitor-2 (CI-2) Gene of Barley

Barley chymotrypsin inhibitor-2 （CI-2） was considered to be a promising candidate for enhancing the nutritional value of other cereals by increasing its concentration as it is rich in lysine than any other storage protein. Also, it was proposed that CI-2 might play an important role in the inhibition of proteolytic enzymes from pests or pathogens as CI-2 can strongly inhibit chymotrypsin and subtilisin. In this study, a total of 93 CI-2 gene sequences were isolated from wild and cultivated barley. 48 SNPs and 4 indels were detected across the entire sequences. The frequency of SNPs in the noncoding region （1 out of 9 bases） was slightly higher than that in the coding region （1 out of 10.7 bases）. In all, 33.3% of the candidate cSNPs resulted in amino acid changes. As a total, the 24 cSNPs resulted in 15 amino acid changes. Ten distinguishable haplotypes were detected, among which 3 haplotypes were shared in the most barley accessions, whereas the rest of the haplotypes appeared at a lower frequency. In addition, three haplotypes （haplotype 4, 8, and 9） were unique for single accessions. These results suggested that low diversity at the CI-2 locus was detected among the cultivated and wild barley.

Kinetic Resolution of DL-Phenylalanine Methyl Ester by α -Chymotrypsin ImmobiIized on Porous Silica Beads

Kinetic resolution of DL-phenylalanine methyl ester was carried out using drimobilized α -chymotrypsin (IC) as catalyst. The effects of temperatUre, pH, concentration of substrate and reactionvessels on the resolution were investigated. High quality L-phenylalanine was obtained in good yieldby an IC column.

PREPAKATION OF THERMALLY REVERSIBLE HYDROGELS AND THEIR USE FOR THE IMMOBILIZATION OF CHYMOTRYPSIN

A series of hydrogels with lower critical solution temperature (LCST) were prepared and used for the immobilization of chymotrypsin. The activities of the immobilized enzyme decrease when the temperatures are raised above their LCST and recover below their LCST. This property is reversible.

<i>Fusarium lini</i>Potential for the Biotransformation of Norandrostenedione and Evaluation of Urease and Chymotrypsin Properties of the Transformed Products

Several androgenic steroids have been biotransformed by fungi into metabolites with numerous biological properties. Incubation of norandrostenedione (1) with Fusarium lini NRRL 2204 was carried out for the first time, yielding two new metabolites, 3,7β-dihydroxy-19-norandrost-1,3,5-trien-17-one (3) and 6α,10β,17β-trihydroxy-19-nor-4-androsten-3-one (4), along with three known compounds, 3-hydroxy-19-norandrost-1,3,5-trien-17-one (2), 10β, 17β-dihydroxy-19-nor-4-androsten-3-one (5) and 10β-hydroxy-19-nor-4- androsten-3,17-dione (6). Their structures were elucidated by extensive spectroscopic analyses, including 1D-, 2D-NMR, and HR-MS experiments. Substrate 1 and its derivatives 2-6 were evaluated in vitro for their urease and chymotrypsin inhibitory properties. Compounds 2 and 3 were found to have strong urease activity with IC50 = 23.7 ± 0.17 and 10.2 ± 0.28 μm, respectively, as compared to the standard drug thiourea (IC50 = 21.6 ± 0.12 μm). Compounds 4, 5 and 6 showed good chymotrypsin activity with IC50 values of 6.4 ± 0.19, 15.6 ± 0.46 and 18.4 ± 0.65 μm, respectively, as compared to standard chymostatin with IC50 = 5.7 ± 0.14 μm. These transformed metabolites may form the basis for the future development of new drugs against ulcer, inflammation, bacterial and viral diseases.

Comparison of the Contributions of Tetrahydrofurfuryl Alcohol and PEG to a-Chymotrypsin Renaturation with High Performance Hydrophobic Interaction Chromatography

The contributions of tetrahydrofurfuryl alcohol (THFA) and polyethylene glycol (PEG) to the renatured efficiency of a-chymotrypsin were investigated and compared with each other. The maximum increments of bioactivity recovery of a-Chy were found to be 25.1% for THFA, 10.4% for PEG, respectively. The experimental results indicated that the denaturant solution containing THFA contributed more to the renaturation of a-Chy in high performance hydrophobic interaction chromatography (HPHIC) than that containing PEG, when the concentration of THFA was 3.2%, the bioactivity recovery of a-Chy is the highest.

Adsorption of α-Chymotrypsin on Plant Biomass Charcoal

The adsorption of α-chymotrypsin onto plant biomass charcoal (PBC), which was prepared from plant biomass wastes such as bagasse and dumped adzuki beans by pyrolysis, has been examined. The PBC was characterized by SEM, specific surface area, and pore size distribution. The adsorption isotherms were successfully correlated by the Freundlich equation. The amount of α-chymotrypsin adsorbed on PBC was dramatically dependent upon the solution pH and temperature. Maximum adsorptions of α-chymotrypsin on adzuki bean charcoal and bagasse charcoal were observed at weak acidic and near neutral pH, respectively. The amount of α-chymotrypsin adsorbed on PBC decreased with an increase in the concentration of salts. Plots of the amount of α-chymotrypsin adsorbed on PBC versus temperature exhibited an optimum temperature.

Heat-Resistant Properties of <i>&#945</i>-Chymotrypsin Adsorbed onto Biomass Charcoal Powder

Biomass charcoal powder (BCP) was used as a carrier for immobilization of α-chymotrypsin through adsorption. BCP was derived from plant biomass wastes such as dumped bamboos by oxygen-free pyrolysis at low temperatures and grinding with a jet mill. The activity of adsorbed α-chymotrypsin was strongly dependent upon the kind of BCP. The thermal denaturation curve of adsorbed α-chymotrypsin was shifted to high temperature, compared to that of free one. When α-chymotrypsin adsorbed onto BCP of bamboos was incubated at 45°C, the half-life of adsorbed α-chymotrypsin was 2.6 times greater than that of free one. After incubation at 45°C, the remaining activity of adsorbed α-chymotrypsin markedly depended on the kind of BCP.

A non-peptide-based chymotrypsin-targeted long-wavelength emission fluorescent probe with large Stokes shift and its application in bioimaging

As a hydrolase,chymotrypsin(CHT)is involved in many physiological activities,and its abnormal activity is closely related to diabetes,pancreatic fibrosis,chronic pancreatitis and pancreatic cancer.In this work,an innovative long-wavelength emission fluorescent probe TCF-CHT was designed and synthesized for the high specificity detection of CHT,which utilized TCF-OH and a mimetic peptide substrate 4-bromobutyryl as chromogenic group and recognition group,respectively.TCF-CHT exhibited excellent selectivity and eye-catching sensitivity(8.91 ng/m L)towards CHT,“off-on”long-wavelength emission at 670 nm and large Stokes shift(140 nm).Furthermore,the successful fulfillment and perfect performance in imaging endogenous CHT in complex organisms(P815 cells,HepG2 cells,zebrafish and tumor-bearing mice)verified its potential as a powerful tool for the recognition of CHT in complicated biological environments.

Chromosomal localization of silkworm (Bombyx mori) sericin gene 1 and chymotrypsin inhibitor 13 using fluorescence in situ hybridization

The chromosomal locations of two single-copy genes, Ser-1 and CI-13, in silkworm (Bombyx mori) were detected at the molecular cytogenetics level by fluorescence in situ hybridization in the study. The results showed that Ser-1 is located near the distal end of the 11th linkage group, relatively at the 12.5±1.4 position in pachytene; and that CI-13 has been mapped near the distal end of the 2nd linkage group, relatively at the 8.2±1.2 position in pachytene. Furthermore, their location model map-FISH map on silkworm chromosome was drawn. The FISH technique and its application to silkworm are also discussed in this paper.

Characterization and transcriptional analyses of cDNAs encoding three trypsin- and chymotrypsin-like proteinases in CrylAb-susceptible and CrylAb-resistant strains of sugarcane borer, Diatraea saccharalis

Diatraea saccharalis is a major corn borer pest. Midgut serine proteinases are essential for insect growth and development. Alteration of midgut proteinases is re- sponsible for Bt resistance development in some species. To clone midgut trypsin and chymotrypsin cDNAs and to test if the CrylAb resistance in D. saccharalis is associ- ated with changes in midgut proteinases, total midgut tryptic and chymotryptic activities, cDNA sequences, and gene expressions of three trypsin and three chymotrypsin genes were comparatively examined between Cry 1 Ab-susceptible （Cry 1 Ab- S S） and Cry 1 Ab-resistant （Cry 1 Ab-RR） strains. Full-length cDNAs encoding three trypsin- and three chymotrypsin- like proteinases were sequenced from CrylAb-SS and CrylAb-RR larvae. These cDNAs code for active forms of midgut serine proteinases with all fimctional motifs, includ- ing signal peptide, conserved His-Asp-Ser for the catalytic triad, three pairs of cysteines for disulfide bridge configurations, and conserved substrate specificity determination residues. In general, cDNA and putative protein sequences are highly similar between CrylAb-SS and CrylAb-RR strains, except for a few nucleotide and predicted amino acid substitutions, whose function need to be further clarified. Total trypsin and chymotrypsin activities were also similar between CrylAb-SS and CrylAb-RR strains. Transcriptional levels of the trypsin and chymotrypsin genes had numerical difference between Cry 1 Ab-SS and CrylAb-RR strains, but the difference was not statistically significant. Data suggest that the development of CrylAb resistance in D. saccharalis was not significantly as- sociated with these trypsins and chymotrypsins. Results clarified the role of six midgut proteinases and provided a foundation for continuing examination of potential involvement of other midgut proteinases in Bt resistance development and other important biochemical processes.

Synthesis of 2,3-dihydroquinazolin-4(1H)-ones catalyzed by a-chymotrypsin

We discovered that a-chymotrypsin has a promiscuous ability to catalyze the cyclocondensation of aromatic and aliphatic aldehydes with 2-aminobenzamides to afford the corresponding 2,3-dihydroquinazolin-4（1H）-ones successfully in high yields（90%–98%） under alcohol solvent. The catalytic activity of a-chymotrypsin was evaluated through investigating the temperature, the enzyme loading and the ratio of substrates in the enzyme-catalyzed reactions. The present method proves to be efficient and environmentally friendly in terms of short reaction time, high yield, green catalyst and the clean products obtained without further purification processes.

Chymotrypsin B is not just a digest enzyme but widely expressed in rat tissues

The functions of lysosomal proteolytic enzymes were once believed to be limited to intracellular protein degradation occured within the lysosome. Recent findings suggest

甲酸对丝素蛋白结晶结构转变的影响

结晶结构是丝素蛋白材料表征的重要内容,对丝素蛋白结晶结构的认识和调控是制备高性能材料的基础。甲酸作为一种可溶解蚕丝的优良溶剂,所制备的再生丝素蛋白材料具有优异的性能,其根源与甲酸对丝素蛋白具有促结晶的作用有关。文章对比分析了基于氯化钙-甲酸溶剂溶制再生丝素蛋白膜与基于三元溶剂溶制再生丝素蛋白膜的结晶结构差异,并利用α-糜蛋白酶从基于三元溶剂制备的丝素蛋白水溶液中分离出高结晶部分和低结晶部分,随后分别利用甲酸和乙醇对分离所得高结晶部分和低结晶部分进行处理,对比分析了乙醇和甲酸对其结晶结构转变的影响。结果表明,相对于乙醇处理,甲酸具有更显著的促丝素蛋白结晶的作用,更有利于丝素蛋白从无规卷曲或Silk I结晶态转变为SilkⅡ结晶态。

糜蛋白酶联合特布他林雾化治疗支气管哮喘的临床效果

目的探讨糜蛋白酶联合特布他林雾化治疗支气管哮喘的疗效及其对肺功能、炎性因子、免疫球蛋白水平的影响。方法将80例支气管哮喘患者随机分为对照组40例与观察组40例。2组均予以常规治疗,在此基础上对照组给予特布他林雾化治疗,观察组给予糜蛋白酶联合特布他林雾化治疗。比较2组临床疗效、临床症状消失时间及治疗前后肺功能指标[第1秒用力呼气容积(FEV1)、FEV1/用力肺活量(FVC)、呼出气一氧化氮(FENO)]、血清炎性因子水平[嗜酸细胞阳离子蛋白(ECP)、γ干扰素(IFN-γ)、白细胞介素-4(IL-4)、白细胞介素-1β(IL-1β)、超敏C反应蛋白(hs-CRP)]、免疫球蛋白水平[免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、免疫球蛋白G(IgG)、免疫球蛋白E(IgE)]的变化;观察2组治疗期间不良反应发生情况。结果观察组治疗总有效率较对照组显著增加(P<0.05),症状消失时间较对照组显著缩短(P<0.05)。2组治疗后FENO值及血清ECP、IL-4、IL-1β、hs-CRP、IgE水平均较治疗前显著降低,FEV1、FEV1/FVC值及血清IFN-γ、IgA、IgM、IgG水平均较治疗前显著升高,且观察组治疗后上述指标较对照组治疗后改善更为显著,差异均有统计学意义(P<0.05)。2组治疗期间不良反应发生率比较差异无统计学意义(P>0.05)。结论糜蛋白酶联合特布他林雾化治疗支气管哮喘疗效确切,可在短时间内改善临床症状,促进肺功能恢复,缓解炎性反应,改善免疫功能。

糜蛋白酶联合布地格福治疗AECOPD的效果及其机制

目的探讨糜蛋白酶联合布地格福治疗慢性阻塞性肺疾病急性加重期(AECOPD)病人的效果及其作用机制。方法选取120例AECOPD病人为研究对象,随机分为研究组和对照组,各60例。研究组治疗方式为基础治疗+糜蛋白酶+布地格福,对照组治疗方式为基础治疗+布地格福,疗程均为4周。比较两组治疗前及治疗4周后肺功能指标、血气分析指标、外周血环状RNA-同源域结合蛋白激酶3(circRNA-HIPK3)和黏蛋白5AC(MUC5AC)水平及临床疗效差异。结果两组治疗前肺功能指标第一秒用力呼气容积(FEV1)、用力肺活量(FVC)、FEV1/FVC、呼气峰值流速(PEF)差异无显著意义(P>0.05);研究组FEV1、FVC、PEF治疗前后差值大于对照组,差异有统计学意义(t=2.840~10.626,P<0.05)。两组治疗前血气分析指标pH、PaO_(2)、PaCO_(2)差异无显著性(P>0.05);研究组pH、PaO_(2)、PaCO_(2)治疗前后差值均大于对照组,差异有统计学意义(t=6.331~6.416,P<0.05)。两组治疗前外周血circRNA-HIPK3、MUC5AC水平差异无显著性(P>0.05);研究组外周血circRNA-HIPK3、MUC5AC水平治疗前后差值均大于对照组(t=2.833、3.250,P<0.05)。研究组治疗效果优于对照组(Z=-1.461,P<0.05)。结论糜蛋白酶联合布地格福治疗更有利于改善AECOPD病人肺功能、血气指标,降低外周血circRNA-HIPK3、MUC5AC水平,提高临床治疗效果。

糜蛋白酶灌注联合自体听骨链重建治疗慢性中耳炎的效果

目的观察糜蛋白酶灌注联合自体听骨链重建对慢性中耳炎听力恢复情况的影响。方法选取100例慢性中耳炎患者作为研究对象,按照随机数字表法将患者分为重建组和联合组,每组50例。两组患者均实施自体听骨链重建术治疗,联合组在此基础上经鼓膜下置管灌注糜蛋白酶配合治疗,比较两组患者的炎症因子、免疫指标、听力恢复情况及并发症发生情况。结果治疗后,联合组的肿瘤坏死因子-α(TNF-α)、降钙素原(PCT)、白细胞介素-8(IL-8)的水平均低于重建组,差异有统计学意义(P<0.05);联合组T淋巴细胞亚群CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)的水平均高于重建组(P<0.05);联合组的气导阈值、气骨导差值均低于重建组,而静态声顺值高于重建组(P<0.05);联合组的纯音听阈值、咽鼓管功能障碍症状评分(ETDQ-7)均低于重建组(P<0.05);联合组的并发症发生率低于重建组(P<0.05)。结论经鼓膜下置管灌注糜蛋白酶联合自体听骨链重建能减轻慢性中耳炎患者的炎症反应,并增强免疫功能,对促进患者听力恢复,降低并发症发生风险均有积极意义。

糜蛋白酶在化脓性伤口治疗中的临床应用效果分析

目的 探讨糜蛋白酶在化脓性伤口治疗中的临床应用效果。方法 选取180例伤口化脓感染患者,按照换药方式不同随机分为观察组和对照组,每组90例。对照组采用常规方法换药后使用生理盐水浸湿无菌纱布湿敷患处;观察组在常规方法换药后使用糜蛋白酶浸湿无菌纱布湿敷患处。比较两组患者伤口愈合时间、换药次数、换药总费用及伤口疼痛程度。结果 观察组伤口愈合时间(15.0±3.8)d短于对照组的(23.0±5.9)d,换药次数(10±2)次少于对照组的(14±3)次,换药总费用(515.8±165.5)元少于对照组的(825.6±205.3)元(P<0.05)。治疗后,两组患者视觉模拟评分法(VAS)评分均低于治疗前,且观察组VAS评分(2.32±1.34)分低于对照组的(3.50±1.25)分(P<0.05)。结论 糜蛋白酶在化脓性伤口治疗中具有较好临床效果,有助于促进切口愈合,减少换药次数,减轻患者经济负担及痛苦,值得临床推广。

0～3周肉仔鸡消化道酶发育规律的研究

本试验以０～３周Ａｖａｉｎ商品代肉仔鸡公雏为研究对象，测试０，４，７，１０，１４，１７和２１日龄肉仔鸡体重，消化道器官重量，胰腺和肠道食糜中淀粉酶、胰蛋白酶、糜蛋白酶和脂肪酶活性的变化，基本摸清了０～３周肉仔鸡体重、消化器官的生长规律及消化道酶的发育规律。试验结果表明：（１）肉仔鸡在４～１０日龄日均相对生长率达到最大值（平均每天增重达１９％），１０日龄后下降，到１４～２１日龄，基本维持在１３％～１４％左右；（２）各器官的相对重量（ｇ／ｋｇ体重）达到峰值的时间并不一致，腺胃和肌胃的相对重量在４～７天为最高，肝和胰在７～１０天，小肠在７天左右；（３）从异速生长的指标看，肌胃的最大增长倍数是体重增长倍数的１．５倍，腺胃是２倍，肝脏是２．３倍，小肠是３倍，胰腺达６倍；（４）小肠内容物的相对重量（ｇ／ｋｇ体重）在４～１０日龄达到最大值，以后呈下降趋势；（５）胰腺和肠道的消化酶活性（活性单位／ｋｇ体重）基本上随着日龄的升高而上升。在胰腺，淀粉酶、胰蛋白酶、糜蛋白酶和脂肪酶活性均在１０，７，２１和１０日龄达到峰值。在肠道，淀粉酶、胰蛋白酶、糜蛋白酶和脂肪酶活性均在１０日龄达到峰值。从消化道酶的分泌情况看，肉仔鸡早期消化酶?

介孔分子筛SBA-15中α-胰凝乳蛋白酶组装及催化活性研究

The hexagonal silica mesoporous material SBA-15 with ordered 2-dimensional channel structure was used as a suppport to the immobilization of emzyme. α-Chymotrypsin, an enzyme, was assembled in the channel of SBA-15 by immersion method. The absorbed amount of enzyme in SBA-15 and the activity of the immobilized enzyme were studied. It was found that there was more absorbed enzyme in SBA-15 than in other carrier and the activity of the immobilized α-chymotrypsin was increased compared with the free enzyme. It suggests that silica mesoporous material SBA-15 is a good support for immobilization of enzyme.