Analytical problems in determining peptide nitrogen in the humic fractions of three Italian soils     

A. Ramunni  R. Scialdone  D. Felleca 《Organic Geochemistry》1985,8(1):87-93

Peptides were released from organic matter fractions of three Italian soils (humin, humic and fulvic acids), when the samples were hydrolyzed in Ba(ON)₂-saturated solution at 105°C for 2 hr. The peptides obtained were separated using electrophoresis and paper chromatography. The presence of polypeptides in the soil organic matter was indicated by: (1) their hydrolysis by pronase; (2) the amino acids released by 6 N HCl hydrolysis; (3) The comparison of i.r. spectra of humic fractions before and after hydrolysis with 6 N HCl.Attempts at isolating the native proteinaceous compounds using electrophoresis in polyacrylamide gel failed; additionally, our attempts to hydrolyze proteinaceous components enzymatically in unfractionated soil organic matter, as well as in its fractions, before and after methylation, with pepsin, papain and pronase, were unsuccessful. Pronase demonstrated a weak proteolytic activity only at very low substrate-enzyme ratios (20 : 1) in humic and fulvic fractions and in whole phyrophosphate extract. Deproteinated substrates treated with pronase also released free amino acids, suggesting autodigestion.In humin, humic and fulvic fractions we found a total amino acid content of 40–45%, 12–24% or 1–85, respectively. Amino acid recovery from single fractions was about 70–80% of the total content in the unfractionated soil.  相似文献   

Production of the Blood Pressure Lowing Peptides from Brown Alga （ Undaria pinnatifida ）     

Sato Minoru Oba Takashi Hosokawa Takao Yamaguchi Toshiyasu Nakano Toshiki Saito Tadao Muramoto Koji Kahara T akashi Funayama Katsura Kobayashi Akio Nakano Takahisa 《中国海洋大学学报(英文版)》2005,4(3):209-213

Brown alga （ Undaria pinnatifida） was treated with alginate lyase and hydrolyzed using 17 kinds of proteases and the inhibitory activity of the hydrolysates for the angiotensin-I-converting enzyme （ACE） was measured. Four hydrolysates with potent ACE-inhibitory activity were administered singly and orally to spontaneously hypertensive rats （SHRs）. The systolic blood pressure of SHRs decreases significantly after single oral administration of the brown alga hydrolysates by pro- tease S ＇ Amano＇ （from Bacillus stearothermophilus） at the concentration of 10 （mg protein） （kg body weight）^ - 1. In the 17 weeks of feeding experiment, 7-week-old SHRs were fed standard diet supplemented with the brown alga hydrolysates for 10 weeks. In SHRs fed 1.0 and 0.1% brown alga hydrolysates, elevating of systolic bloodpressure was significantly suppressed for 7 weeks. To elucidate the active components, the brown alga hydrolysates were fractionated by 1-butanol extraction and HPLC on a reverse-phase column. Seven kinds of ACE-inhibitory peptides were isolated and identified by amino acid composition analysis, sequence analysis, and LC-MS with the results Val-Tyr, Ile-Tyr, Ala-Trp, Phe-Tyr, Val-Trp, Ile-Trp, and Leu-Trp. Each peptide was determined to have an antihypertensive effect after a single oral administration in SHRs. The brown alga hydrolysates were also confirmed to decrease the blood pressure in humans.  相似文献   

益气活血方对射血分数保留型心力衰竭患者血清BNP、hs-CRP及心功能的影响          下载免费PDF全文

顾亚威  莫云秋 《应用海洋学学报》2021,37(6):1

目的：观察益气活血方对射血分数保留型心力衰竭（HFpEF）患者进行干预后,其心功能、脑钠肽（BNP）、超敏C反应蛋白（hs-CRP）的变化情况。方法：将84 例HFpEF患者按照随机数字表法分为治疗组和对照组,每组各42 例。对照组采用西医标准化治疗方案,治疗组在对照组基础上给予益气活血方治疗,2组疗程均为4周。观察2组治疗前后心功能指标［包括心室收缩末期内径（LVESD）、左心室舒张末期内径（LVEDD）、左室射血分数（LVEF）、6 min步行试验（6MWT）］、BNP及hs-CRP的变化,并评价综合疗效。结果：总有效率治疗组为90.48%（38/42）,对照组为80.95%（34/42）,2组比较,差异有统计学意义（P＜0.05）。治疗后2组LVESD、LVEDD及血清BNP、hs-CRP水平均较治疗前降低,LVEF较治疗前升高,6MWT较治疗前增加,且治疗组各项指标改善情况均优于对照组,差异均有统计学意义（P＜0.05）。结论：益气活血方能够抑制HFpEF患者体内BNP和hs-CRP的表达,改善心功能指标,增加6MWT距离。  相似文献   

中药治疗便秘型肠易激综合征SD大鼠的实验研究          下载免费PDF全文

高鸿智  杨勇军  刘仕鸿 《应用海洋学学报》2021,37(4):161

目的：研究柴胡疏肝散合济川煎对肝郁气滞合并阳虚型便秘型肠易激综合征（IBS-C）模型大鼠的治疗效果及作用机制。方法：将56只SPF级雄性SD大鼠随机分为正常组（8只）和模型组（48只）,模型组采用冰水灌胃、限制饮水结合慢性束缚应激的方法制备肝郁气滞合并阳虚型IBS-C大鼠模型。将造模成功的大鼠随机分为模型对照组、复方聚乙二醇电解质散（PEG-EP)组、黛力新组及中药低、中、高剂量组,每组各8只。各组进行药物干预14 d。观察指标包括大鼠一般状态、糖水偏好率、旷场实验评分、内脏敏感性、小肠推进率、结肠黏膜HE染色及血清5-羟色胺( 5-HT) 、生长抑素(SS) 、血管活性肠肽（VIP）含量测定。结果：与模型对照组相比,柴胡疏肝散合济川煎能使模型大鼠的粪便粒数、粪便含水量、Bristol 分值、内脏敏感性、旷场试验评分、糖水偏好百分比、小肠推进率增加(P<0.01 或P<0.05),血清5-HT、SS含量降低(P<0.05),血清VIP含量升高（P<0.05）;与PEG-EP组、黛力新组相比,中药高剂量组大鼠血清5-HT、SS均降低(P<0.05),VIP水平升高(P<0.05)。结论：大鼠肝郁气滞合并阳虚型IBS-C模型制备成功,柴胡疏肝散合济川煎可改善模型大鼠肝郁和便秘症状,其作用机制可能与降低大鼠血清5-HT、SS含量和升高大鼠血清VIP含量有关。  相似文献   

响应面法优化酶法制备藻蓝蛋白抗氧化肽的实验研究     

唐志红  焦绪栋  周云丽  周虹  吕家森  秦松 《海洋科学》2012,36(11):50-54

为优化碱性蛋白酶酶解藻蓝蛋白制备抗氧化肽的工艺条件,采用响应面分析法,以超氧阴离子自由基清除率为响应值,研究了[E]/[S]、酶解温度和时间对制备抗氧化肽工艺的影响.结果表明,制备藻蓝蛋白肽的最佳酶解条件为:温度为44.9℃、时间为6.4 h和[E]/[S]为3.6%,此时对超氧阴离子自由基清除率达到75.39%.在该条件下制备的藻蓝蛋白酶解产物(0.5 g/L)还原能力的吸光度为1.03,并达到稳定.  相似文献   

Mycosynthesized Silver Nanoparticles as Potent Growth Inhibitory Agents Against Selected Waterborne Human Pathogens     

《洁净——土壤、空气、水》2017,45(4)

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大菱鲆Hepcidin基因的克隆和真核表达载体的构建     

丛柏林  韩光  黄晓航  刘胜浩  林学政  陈靖 《海洋科学进展》2008,26(1):74-81

Hepcidin是鱼类的一种重要的抗菌肽.通过比较GenBank中不同Hepcidin基因的同源相似性,设计出特异性的引物,以我国海水养殖大菱鲆(Scophthalmus maximus L)肝脏为模板利用RT-PCR 技术克隆获得了一条基因.使用生物信息学手段对该基因序列进行了比对和分析,证实这条基因为Hepcidin1 precursor基因,属于Hepcidin基因家族.通过设计引物在Hepcidin基因的5′和3′端分别引入EcoR I和NotⅠ酶切位点.经过双酶切后获得具有EcoR I和Not I酶切位点的Hepcidin成熟肽片段,并与同样使用该酶线性化的表达载体pPIC3.5K连接.构建了表达载体pPIC3.5K/Hepc1,并将其导入到酵母基因组中,完成了真核表达载体的构建,为Hepcidin的真核表达作了基础性的准备工作.  相似文献   

Screening of protease-producing marine yeasts for production of the bioactive peptides   总被引：1，自引：0，他引：1  

NI Xiumei  CHI Zhenming  LIU Zhiqiang  YUE Lixi 《海洋学报(英文版)》2008,27(2):116-125

Over 400 yeast strains from seawater and sediments were obtained, but only five strains named HN2 -3, N13d, N13C, Mb5 and HN3 - 2 among them could form clear zones around their colonies on the double plates with 2.0% casein. Peptides in the hydrolysate produced by the proteases from strains HN2 -3 and N13d had higher angiotensin I-converting-enzyme （ACE）-inhibitory activity. The two marine yeast strains were identified to be Aureobasidium pullulans according to the results of routine yeast identification and molecular methods. After purification of the proteases from the two marine yeast strains, it was found that the optimal pH for them was both 9.0, both of them were serine alkaline protease. However, the optimal temperature for the protease from the strain HN2 -3 was 52℃ while that from strain N13d was 48℃. ACE-inhibitory activity of the peptides in the hydrolysate of shrimp protein produced by the purified protease from the strain HN2 -3 was the highest while antioxidant activity in the hydrolysate of spirulina protein produced by the purified protease from the strain N13d was the highest.  相似文献   

Role of shellfish hatchery as a reservoir of antimicrobial resistant bacteria     

Claudio D. Miranda  Rodrigo Rojas  Marcela Garrido  Julieta Geisse  Gerardo González 《Marine pollution bulletin》2013

The main aim of this study was to determine the occurrence of resistant bacteria in florfenicol-treated and untreated scallop larval cultures from a commercial hatchery and to characterize some selected florfenicol-resistant strains. Larval cultures from untreated and treated rearing tanks exhibited percentages of copiotrophic bacteria resistant to florfenicol ranging from 0.03% to 10.67% and 0.49–18.34%, respectively, whereas florfenicol resistance among oligotrophic bacteria varied from 1.44% to 35.50% and 3.62–95.71%, from untreated and treated larvae, respectively. Florfenicol resistant microbiota from reared scallop larvae mainly belonged to the Pseudomonas and Pseudoalteromonas genus and were mainly resistant to florfenicol, chloramphenicol, streptomycin and co-trimoxazole. This is the first study reporting antimicrobial resistant bacteria associated to a shellfish hatchery and the results suggest that a continuous surveillance of antimicrobial resistance even in absence of antibacterial therapy is urgently required to evaluate potential undesirable consequences on the surrounding environments.  相似文献   

中国对虾素在大肠杆菌(Escherichia coli)中的重组表达   总被引：3，自引：0，他引：3  

薛剑峰  康翠洁  王金星  赵小凡  相建海 《海洋与湖沼》2005,36(3):235-240

中国对虾素是从中国对虾血细胞中克隆得到的一种抗菌肽。为了进一步研究中国对虾素(CHP)的功能并为制备特异性抗体作准备，采用大肠杆菌表达外源蛋白的方法，进行了对虾素原核表达的研究。根据从中国对虾血细胞中克隆得到的对虾素基因设计特异性引物扩增中国对虾素成熟肽基因(CHP)，插入原核表达载体pGEX-4T-1中，在E．coli BL21表达融合蛋白GST-CHP．结果表明，不同表达菌株的融合蛋白表达量为30％—34％，同一菌株在诱导5h后能达到最高表达量。利用GST亲和层析纯化融合蛋白，将融合蛋白用凝血酶裂解以得到CHP，其分子量约为5．6kDa，N-端测序结果与期望的成熟对虾肽序列一致。用液体生长抑制方法检测活性，表明重组GST-CHP蛋白及CHP均表现出对大肠杆菌的抑菌活性。  相似文献