Improved elastase production by Bacillus sp. EL31410—further optimization and kinetics studies of culture medium for batch fermentation

An efficient culture medium producing a bacterial elastase with high yields was developed further following preliminary studies by means of response surface method. Central composite design (CCD) and response surface methodology were applied to optimize the medium constituents. A central composite design was used to explain the combined effect of three medium constituents, viz, glucose, K(2)HPO(4), MgSO(4).7H(2)O. The strain produced more elastase in the completely optimized medium, as compared with the partially optimized medium. The fitted model of the second model, as per RSM, showed that glucose was 7.4 g/100 ml, casein 1.13 g/100 ml, corn steep flour 0.616 g/100 ml, K(2)HPO(4) 0.206 g/100 ml and MgSO(4).7H(2)O 0.034 g/100 ml. The fermentation kinetics of these two culture media in the flask experiments were analyzed. It was found that the highest elastase productivity occurred at 54 hours. Higher glucose concentration had inhibitory effect on elastase production. At the same time, we observed that the glucose consumption rate was slow in the completely optimized medium, which can explain the lag period of the highest elastase production. Some metal ions and surfactant additives also affected elastase production and cell growth.     

Homocysteine increases the synthesis of adrenomedullin in vascular fibroblasts of rats

Homocysteine (Hcy), a highly reactive sulfur- containing amino acid, is an intermediate product of methionine metabolism. Elevation of plasma Hcy has been widely studied as an independent risk factor for many cardiovascular diseases such as atherosclerosis, thrombosis, etc.[1]. Inherited hyperhomocysteinemia cases usually suffer from occlusion or thrombosis in arteries and large veins before the age of 20. Hyperhomocysteinemia significantly reduces the survival rate[2] and dilatation functi…     

人转铁蛋白基因的克隆及序列分析

目的:克隆人转铁蛋白基因并对其编码序列进行分析.方法:以人胎肝cDNA为模板,利用PCR方法克隆人转铁蛋白基因;通过与基因组序列对比分析基因组结构;通过TargetP 1.1和SignalP 3.0预测信号肽;通过Clustal X(1.81)进行蛋白序列联配.结果:PCR扩增了一个长2 160 bp的基因片断,序列分析表明其覆盖了完整编码框,编码由698个氨基酸组成的人转铁蛋白.进一步分析发现人转铁蛋白基因有19个外显子和18个内含子,编码人转铁蛋白N端具有19个氨基酸组成的信号肽序列.人转铁蛋白与猩猩、猴子、兔子和老鼠的转铁蛋白氨基酸相似率分别为94%、91%、78%、73%.生物信息分析表明,人转铁蛋白含有高度保守的参与蛋白二硫键形成的半胱氨酸以及铁离子结合位点,有两个序列较同源的结构域.结论:成功克隆人转铁蛋白基因,人转铁蛋白与其它物种转铁蛋白同源.     

培养条件对康宁木霉产酶的影响

通过对康宁木霉纤维素酶（C1酶和Cx酶）、酸性蛋白酶活力的比较,筛选出了康宁木霉产酶的最适培养条件.康宁木霉在m（麸皮）∶m（秸秆）=2∶8,氮源为w=1%的NH4NO3,含水量为65%,起始pH值为6.5的条件下培养72h,C1酶活力最高,为191.3U/g;在m（麸皮）∶m（秸秆）=6∶4,氮源为w=1%的（NH4）2SO4,含水量为65%,起始pH值为7.5的条件下培养48h,Cx酶活力最高,为2006.5U/g;在m（麸皮）∶m（秸秆）=8∶2,氮源为w=1%的（NH4）2CO3,含水量为65%,起始pH值为4.5的条件下培养96h,酸性蛋白酶活力最高,为1859.6U/g.     

鲤鱼肌肉中胶原蛋白酶的分离纯化与性质分析

利用明胶酶谱法,在鲤鱼肌肉中检测到了一种具有分解胶原蛋白能力的丝氨酸蛋白酶.通过硫酸铵盐析、DEAE-Sephacel和Phenyl-Sepharose等一系列柱层析法,纯化得到了分子质量为70 ku,能够降解胶原蛋白的丝氨酸蛋白酶(Collagenolytic Serine Proteinase,CSP).性质分析表明,CSP在弱碱性条件下显示活性,最适温度为30℃;丝氨酸蛋白酶抑制剂特异抑制CSP的活性.CSP能有效地分解鲤鱼肌肉中的I型胶原蛋白,提示其可能在鱼体死后肌肉胶原蛋白降解中发挥着重要的作用.     

采用蛋白酶组合漂白技术对羊毛增白的探讨

根据厂家对羊毛织物白度要求，需要开发一种快速，安全，价廉的羊毛漂白生产技术以满足于生产研究，研究了蛋白关正常氧漂工艺中对织物白度的促进作用及采取正确处理方法是获得良好的必要条件。     

羊毛表面性状的微观研究

借助扫描电镜和透射电镜对洗净澳毛的表面微观结构进行研究。并通过对其鳞片细胞的红外吸收光谱图的化学结构分析。建立了比较完整的羊毛纤维表面微观物理结构和化学结构模型。经过变性处理后，羊毛纤维表面性状结构发生了预期变化。进一步证明了毛纱上浆率低的根本原因是表面疏水性物质的存在。     

碱性蛋白酶催化的Henry反应

Henry反应是一类重要的碳碳键形成反应，广泛用于各类天然产物和医药中间体合成中．使用碱性蛋白酶催化芳香醛和硝基甲烷之间的Henry反应，在室温的条件下，得到对应的产物，产率高达88％．     

亚硝酰铁氰化纳法测定复杂样品中半胱氨酸含量的方法研究

本文首次建立了亚硝酰铁氰化钠法测定半胱氨酸的方法，该法灵敏度高，选择性好，干扰少，线性范围为１－９０μｇ／ｍＬ，检出极线为０．７８μｇ／ｍＬ，回归方程为ｙ＝０．０１７６ｘ＋０．２１１４，相关系数ｒ＝０．９９９１，尤其适用于复杂样品中半胱氨酸含量的测定。     

高度顺/反选择性合成单氟烯烃

近年来，单氟烯烃在药物化学中得到了广泛关注。这主要是由于单氟烯烃骨架在电荷分布与偶极距性质上与酰胺结构非常相近，但是单氟烯烃不像酰胺键那样容易发生构象变化或在水解酶的作用下发生水解断裂。因此单氟烯烃结构单元作为仿酰胺结构，常被用于仿肽类（peptidomimetic）蛋白酶抑制剂类药物中。