基于Access和Dream Weaver软件的实验室菌种管理系统设计与应用

利用开放性实验和实践创新训练的教学模式为学生提供实践机会,指导学生参与实验室中的基因工程菌菌种管理。针对实验室构建的工程菌的品种多、数量大、流动快等特点,利用Microsoft Office Access2003和Macromedia Dream Weaver 8.0设计了一种简单易行的菌种管理系统,对现有菌种进行良好的统计和查找。阐述了如何使用Microsoft Office Access 2003和Macromedia Dream Weaver 8.0软件管理实验室菌种,并介绍了该系统的实际应用。     

固定化脂肪酶合成蔗糖月桂酸酯的研究

以固定化脂肪酶为催化剂,以DMSO/叔丁醇有机溶剂为反应体系,在室温条件下催化合成蔗糖酯。研究表明:月桂酸与蔗糖的摩尔比为大于8,反应温度为30℃,脂肪酶用量为0.3g,缓冲液的pH值为7.5,反应6h时,酯化反应的相对转化率较高。反应产物的薄层色谱和红外光谱分析结果证实所合成的产物为蔗糖月桂酸酯。     

漆酶高产菌株的筛选

采用平板PDA加愈创木酚的方法,对30株白腐菌和10株食用菌进行产漆酶初筛和复筛,得到了10株高产菌株,其中酶活最高的菌株为灵芝(Glossy ganoderma sp.),最高产酶活可达到478.5U/L.     

两相体系脂肪酶催化合成蔗糖酯的研究

研究了正戊醇和水形成的两相体系混合溶剂中,蔗糖和乙酸乙烯酯在游离态脂肪酶的催化作用下,发生转酯化反应合成蔗糖酯,确定了合成反应的各工艺参数。即当蔗糖为1.0g,水体积含量为60%,温度在40℃,缓冲液的pH值为8.0,脂肪酶用量为0.3g,蔗糖与乙酸乙烯酯的比例为1∶4,反应时间24h时,蔗糖的转化率为54.8%。通过红外光谱确定了生成主要的产物是蔗糖-6-乙酸酯。     

一次性运动对肥胖大鼠脂肪水解调节因素的影响

研究一次性运动对肥胖大鼠脂肪组织HSL、perilipin蛋白含量及脂肪水解的影响。方法:116只3周龄SD雄性大鼠,110只给予10周高脂饲料,建立高脂饮食诱导肥胖大鼠模型。随机分为:安静对照组,运动后即刻、1、31、2和24 h组。用化学法测试血清FFA和甘油、脂肪组织HSL活性;用Western Blotting法测试HSL和perilipin蛋白表达。结果:与对照组相比,运动后FFA和甘油、HSL活性和含量出现不同程度升高;而perilipin含量除即刻组出现显著升高外,其它各组均显著降低。结论:一次性运动导致肥胖大鼠运动后脂解增加,可能与其脂肪组织HSL活性、HSL蛋白含量上调及perilipin蛋白含量下调有关。     

无溶剂体系中脂肪酶催化合成癸酸甘油酯

研究了无溶剂体系中固定化脂肪酶催化癸酸和甘油合成癸酸甘油酯，并与油酸进行对比。癸酸和油酸分别在50℃和60℃下获得最高平衡转化率。在开口反应器中，加酶量100u／g，50℃下癸酸最高平衡转化率为90％，移去水的方不同对最终转化率影响很大。     

非平衡双轴失配应变对BST薄膜相图的影响

用朗道———德文希尔理论研究了非平衡双轴失配应变对单畴外延钛酸锶钡(BST)薄膜相变的影响,得到了室温下生长在四方基底上的BST50/50薄膜的“失配应变—失配应变”相图和能量图。结果表明在BST50/50薄膜中由于非平衡双轴失配应变的作用,出现了两个在平面内的四方铁电相,即:a1相(P1≠0,P2=P3=0)和a2相(P2≠0,P1=P3=0),而这在平衡失配应变下是不存在的。室温下,非平衡失配应变在一定范围内不能使BST50/50中的顺电相消失。     

Sex Dimorphism in Serum Lecithin: Cholesterol Acyltransferase and Lipoprotein Lipase Activities in Adult Sickle Cell Anaemia Patients with Proteinuria

Proteinuria in subjects with sickle cell anaemia (SCA) is an indication of an ongoing renal insufficiency and it’s prevalence varies between sexes. We evaluated sex differences in the activities of Lecithin: cholesterol acyltransferase (LCAT), Lipoprotein lipase (LPL) and the levels of lipoproteins in SCA patients with proteinuria. Fifty SCA patients (30 males aged: 26.4 ± 7.3 years and 20 females, aged 25.4 ± 2.6 years) and 50 age and sex matched control SCA patients were recruited for the study. Random urine specimens were collected and tested for the presence of albumin by urine dipstick technique. A 24 h urinary protein was quantitated using sulphosalicylic acid technique. Fasting serum total cholesterol, triglyceride, urea and creatinine were determined using enzymes catalyzed colorimetric methods. HDL cholesterol was determined in the supernatant after precipitation with manganese chloride–phosphotungstic acid solution. LCAT was measured using the Anasolv LCAT assay with proteoliposome as substrate. LPL was determined by incubating the serum in glyceryl trioleate substrate, the glycerol liberated was measured in an aliquot of the incubating mixture. In male SCA controls there was 18.2 and 6.9% increase in the activities of LPL and LCAT respectively when compared with females but in SCA patients with proteinuria there was 8.4 and 5.2% decreases in the male SCA patients compared with females. The concentration of 24 h urine protein in the SCA male subjects with proteinuria was significantly higher (0.25 g/day; P < 0.001) compared with the SCA female patients with proteinuria (0.09 g/day). There are sex differences in the activities of LCAT and LPL in SCA patients with proteinuria. Metabolism of these lipolytic enzymes may be modulated differently in SCA patients with proteinuria.     

丁草胺与胡子鲶肝胰脏淀粉酶及脂肪酶活力

本实验研究3种不同浓度(0.1、1.0、2.5 mg/L)的丁草胺对胡子鲶肝胰脏淀粉酶、脂肪酶活力的影响.结果表明,暴露2 d的高浓度组(2.5 mg/L)及暴露4 d的各浓度组淀粉酶活力显著提高,暴露6d的低浓度组(0.1 mg/L)及较高浓度组(1.0 mg/L)仍保持激活特性,而高浓度组(2.5 mg/L)淀粉酶活力则受到显著抑制;丁草胺暴露期间胡子鲶肝胰脏脂肪酶活力始终呈现显著的抑制作用,特别是高浓度组(2.5 mg/L)随着暴露时间的延长(6 d)则呈现极显著抑制.     

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtilis was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone α-factor (MFals), and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction (PCR)-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae (SC-βG) was preliminarily screened by the clearing hydrolysis zone formed after the barley β-glucan was hydrolyzed in the plate and no proteinase A (PrA) activity was measured in fermenting liquor. The results of PCR analysis ofgenome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-1,3-1,4-β-glucanase assay methods showed that the recombinant strain SC-βG had high endo-1,3-1,4-β-glucanase expression level with the maximum of 69.3 U/(h-ml) after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-1,3-1,4-β-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.