The use of limited proteolysis to probe interdomain and active site regions of beta-galactosidase (Escherichia coli)

Limited proteolysis by pancreatic elastase (EC 3.4.21.36) and chymotrypsin (EC 3.4.21.1) was used to study the domain structure and active site of beta-galactosidase (EC 3.2.1.23) (Escherichia coli). Treatment with elastase resulted in a rapid cleavage between residues Ala-732 and Ala-733. No inactivation accompanied this cleavage suggesting that this bond is in a hinge region of the protein. Some slow cleavages beyond the initial one were observed to occur and were accompanied by inactivation. Treatment of beta-galactosidase with chymotrypsin resulted in cleavages first between Trp-585 and Ser-586 and then between Phe-601 and Cys-602. The first of these cleavages resulted in total inactivation of beta-galactosidase. The presence of monovalent ions or isopropyl-beta-D-thiogalactopyranoside protected against the cleavages but when Mg2+ or Mn2+ was present in the reaction mixture, the bond between Trp-585 and Ser-586 was more susceptible to the action of chymotrypsin. These data demonstrate that the conformation of beta-galactosidase around Trp-585 and Ser-586 is dramatically affected by the binding of ions and isopropyl-beta-D-thiogalactopyranoside. The mutant M15 beta-galactosidase, which is missing residues 11 through 41 and is an inactive dimer rather than an active tetramer, was found to be much more labile to proteases than native beta-galactosidase, but the same initial cleavages were found to occur. In addition, trypsin cleaved the M15 protein between Arg-431 and Trp-432 while native beta-galactosidase was stable to trypsin.     

糙叶败酱挥发油镇静作用的研究

本文观察了败酱科植物糙叶败酱(Patrinia scabra Bunge)根和根茎中制得的挥发油的镇静作用,井与黄花败酱挥发油做了比较。结果表明,此油灌胃给予数组小鼠,剂量0.45ml/kg,显示如下的作用:[1]能显著延长由于腹腔注射戊巴比妥钠(50mg/kg)引起的小鼠睡眠时间,但其作用强度弱于黄花败酱挥发油。(2)一次灌胃给予小鼠大剂量10.46g/kg的糙叶败酱挥发油,连续观察10天,动物外观正常,无一死亡,体重增加与对照组相似。     

鲢、鳙在天然条件下的摄食强度(Ⅱ)武汉东湖鲢、鳙周年摄食强度的研究

武汉东湖的鲢(Hypophthalmichthy molitrix)鳙(Aristichthys nobilis)在天然条件下摄食强度具季节性变化。摄食强度高峰处于夏季,低谷处于冬季。在实验条件下,按周年采样期间水温变化范围,测定鱼的肠管排空率。食物通过鱼肠管时间(Y_p—h)与水温(X_t—℃)的关系为: 鲢Y_p=270.63 X_t~(0.6408) 鳙Y_p=280.46 X_t~(0.6642) 根据修正后Bajkov公式(D=C (24.A)/n),估算鱼的日粮。鱼日粮(Y_D)与水温(X_t)关系为: 鲢Y_D=0.2683e~(0.1503X_t) 鳙Y_D=0.0075X_t~(2.2715) 计算鱼在天然条件下周年月粮及年粮。鲢、鳙对天然饵料年消耗量分别为18.924公斤及17.39公斤,饵料系数分别为18.02及13.38。     

“引黄济青”输水沿线水生生物分布与蓄水库富营养化关系的研究

“引黄济青”是国家“七五”重点工程,由黄河下游博兴县境内打渔张闸引水,在渠首沉沙后,再经惠民、东营、潍坊和青岛的10个县区约253km的明渠输送,到达唠山县棘洪滩蓄水库蓄存,实现向青岛市日供水3.0×10~5m~3。明渠输水过程中,来白沿线的营养物质会污染水体,加之输水在库中滞留时间长,会不会发生富营养化,这是社会普遍关注的问题。     

自养黄杆菌合成羟基丁酸和羟基戊酸共聚体的发酵研究

采用本实验室从土壤中分离到的一株自养黄杆菌进行了羟基丁酸和羟基戊酸共聚体〔P(HB-co-HV)〕的发酵试验。实验结果表明,该菌株是自养黄杆菌葡萄糖运输突变株,可以葡萄糖、果糖、蔗糖、麦芽糖、乙酸盐、乳酸盐和苹果酸盐作为唯一碳源,尤以葡萄糖和果糖效果最佳。硫酸铵、氯化铵和蛋白胨等不同氮源不影响其生长,却影响细胞中P(HB-co-HV)的含量和P(HB-co-HV)中HV/HB的比例。应用两阶段控制方式,经42h的补料分批发酵,细胞浓度达34.9g·L~(-1),P(HB-co-HV)浓度达25.28g·L~(-1)。细胞和P(HB-co-HV)生产速率系数分别为0.83g·L~(-1)”·h~(-1)和0.61g·L~(-1)·h~(-1)。以基质为基准的细胞得率系数(Yx/s)、产物得率系数(Yp/s)和以干细胞为基准的产物得率系数(Yp/x)分别为0.283(g/g)、0.174(g/g)和0.73(g/g)。改变培养基中碳氮源组分可将P(HB-co-HV)中HB的含量调节在24％～78％之间。     

湿地松粉蚧夏季数量凋落的原因分析

利用生命表技术及其相应的控制指数分析方法,对新侵入害虫湿地松粉蚧Ｏｒａｏｅｌｌａａ－ｃｕｔａ（Ｌｏｂｄｅｌｌ）夏季数量凋落的某些因子进行了量化分析。结果表明：在广东南部的新侵入区,夏季高温引起的松梢迅速老化,上代为害以后引起的营养质量的变化、拥挤以及煤污病的严重发生等,均对湿地松粉蚧夏季种群数量的凋落有着明显的影响,其总的排除作用控制指数ＥＩＰＣ为４６．８９,如果排除这几个因子的作用,下代种群的数量将为当代的４６．８９倍。     

边界层阻力在叶片气体交换过程中的作用

作者对现有的气体交换法的叶室作了改进。从实验上检验了作者根据电学类比分析和模拟检验得到的结论：在已有的气体交换测定装置中,空气流速不变,叶片的边界层导度也固定不变,所得到的结论用于边界层导度变化的情形时,会造成比较大的误差。现加一障碍物使得流过叶片的风速减小,从而改变叶片的边界层导度。通过测量在不同边界层导度下同一叶片的光强曲线可以考察不同边界层导度下的气孔导度、光合速率、蒸腾速率以及它们之间的相互关系。实验结果与作者以前的电学类比分析和模拟得到的结论是基本一致的