用废啤酒酵母制备碱不溶性葡聚糖的研究

研究了用碱法及酶－碱法从啤酒厂排弃的酵母泥中制备碱不溶性葡聚糖的工艺。对NaOH及蛋白酶用量的影响,NaOH作用时间、浓度、作用次数等因素的影响进行了较系统的探讨。先用蛋白酶处理酵母可减少NaOH用量从而减轻环境污染。对多糖产品进行了定性定量分析,得到的产品主要为β（1→3）-D-葡聚糖。     

原生质体融合技术选育耐热性β-淀粉酶产生菌

从土壤中分离到两株产β-淀粉酶芽孢杆菌菌株,经紫外线、丫-射线,氯化锂、亚硝基胍等诱变和筛选,得到β-淀粉酶高产菌和耐热性β-淀粉酶产生菌各一株。将两菌进行原生质体融合,获得兼有两亲株遗传特性的融合子wg6。从菌落形态和产酶特性等证实此融合子系两亲株融台所得的杂交子代。W96菌株产酶能力介于两亲株之间,酶的热稳定性较高,60℃处理15min,酶活力仍达93．2％。此菌株还可产生少量茁霉多糖酶(一种支链淀粉酶)．与所产生的β-淀粉酶协同作用,使淀粉水解率达到80．6％,因而有较高的应用价值。     

海枣曲霉β-葡萄糖苷酶的催化性质

本文研究了海枣曲霉(Aspergillus phoenicis)β-葡萄糖苷酶的底物特异性以及不同化学试剂对酶活力的影响。该酶水解对一硝基酚基-β-葡萄糖苷、纤维二糖和水杨素的相对活力分别为100、180和67.3。水解对一硝基酚基β-葡萄糖苷和纤维二糖的Km值分别为0.97mmol／L和1 8mmol／L,Vmax分别为576μmol min-1.mg-1和595μmol.min-1.mg-1.mg-1。Ag+、D-葡萄糖和纤维二糖对酶活力有强烈的抑制作用。Lineweaver—Burk作图法及Dixon作图法表明D-葡萄糖对该酶显示竞争性抑制作用,其Ki值分别为30mmol／L和3．4mmol／L。     

生物法合成维生素C棕榈酸酯

研究了不同的脂肪酶在有机溶剂体系中催化合成L-维生素C棕榈酸酯的反应。针对维生素C在有机溶剂中溶解度较低这一问题,对催化合成维生素C棕榈酸酯反应的脂肪酶和反应介质进行比较,同时对影响合成维生素C棕榈酸酯反应的因素（温度、底物浓度、底物摩尔比、反应时间和酶量等）进行探讨,优化了反应条件：在10mL的丙酮中,1.094g棕榈酸与0.107g维生素C在酶量为20％（W/W, 固定化酶/维生素C）的固定化脂肪酶催化下,初始含0.4nm分子筛20%,温度为60℃,转速为200r/min,反应48h转化率可以达到80%,产物维生素C棕榈酸酯的浓度可达20g/L。     

碱性β-甘露聚糖酶发酵工艺的研究

本文报道碱性β-甘露聚糖酶16L罐的发酵工艺。在发酵过程中,通气量影响菌体生长,最适通气量为1:0.75—1:1.0vvm。搅拌速度影响菌体产酶,最适搅拌速度为500r/min。碳源为魔芋粉,其适宜浓度为2%。发酵周期为40小时。发酵液中β-甘露聚糖酶的酶活力达300u/ml,比摇床上培养提高了2倍。     

Paenibacillus massiliensis T7固氮基因簇nifBHDKENX的克隆、序列分析及铵和氧对nifB启动子的调控

Paenibacillus massiliensis T7是我室从北京郊区分离的一株革兰氏阳性、产芽孢的固氮菌. 根据GenBank中已发表的多种固氮生物nifH基因序列, 在其保守区域设计一对简并引物, PCR扩增获得324 bp的nifH片段. 用地高辛标记该片段为探针, 对EcoRⅠ和PstⅠ消化的P. massiliensis T7染色体DNA进行Southern杂交, 结果DNA/EcoRⅠ杂交道1.1 kb和9 kb处, DNA/PstⅠ杂交道1.3 kb和8 kb处分别有两条杂交带, 从而初步判断nifH可能有两个拷贝. 在324 bp nifH片段的基础上, 采用反向PCR扩增的方法, 获得了purD和nifBHDKENX基因. 在nifB的上游有一个类似σ⁵⁴型启动子的保守序列, 在该启动子上游有一个类似激活蛋白NifA结合位点. 分析表明, nifBHDKENX可能组成一个转录单元, 共用nifB上游的启动子. 将nifB启动子与报告基因lacZ相融合, 构建成表达载体pnifB-LacZ, 并转化到P. massiliensis T7中, 研究铵和氧对该启动子的表达调控. 结果表明, 在无氧条件下, 铵浓度为0%时, β-半乳糖苷酶的活性最高, 随着铵浓度的增加, 酶活性并没有明显的下降过程, 即使铵浓度达80 mmol/L时, 酶活也相当于无铵时的73.12%, 说明铵对nifB启动子有微弱的抑制作用, 而铵浓度一定, 氧浓度3%时, β-半乳糖苷酶活性最高, 提高氧浓度则酶活性开始下降, 当氧浓度高于20%时, β-半乳糖苷酶活性仅几个Miller单位, 说明氧对nifB启动子有明显的抑制作用.     

人p53蛋白在巴斯德毕赤酵母中的表

将人p53基因装入Pichia分泌型质粒Phil-S1中,酶切线性化后电穿孔导入酵母细胞进行整合,经筛选得到一高表达p53蛋白的克隆。SDS-PAGE显示表达量约占分泌总量的30%。ELISA验证重组人p53存在免疫学活性。在诱导时就降低Pichia酵母系统水解酶活力等方面进行优化,经FPLC分离纯化得到约200mg/L表达量。     

海枣曲霉β-葡萄糖苷酶的提纯与性质

通过聚乙二醇6000一磷酸钾缓冲液双相分离,Sephadex G—100凝胶过滤、DEAE-Sephadcx A-50及SE-Se phadex C-50离子交换柱层析等提纯步骤,从海枣曲霉(Aspe rgillutphoenlcis)麦麸培养物抽提液中提纯到凝胶电泳均一的β-葡萄糖苷酶。该酶的最适pH5．0,最适温度60℃,在pH 4．0--7．5之间及55℃以下稳定。Ag+及Hg2+对该酶有强烈的抑制作用。用SDS-凝胶电泳法及梯度凝胶电泳法测得该酶均分子量分别为118000及195000薄层凝胶等电聚焦法测得其等电点为pH 3.95。     

青霉素G酰化酶调节基因的定位

为了定位青霉素G酰化酶的调节基因,从质粒Ppa6克隆了一系列青霉索G酰化酶基因(pac)的片段,将这些重组质粒转化E.coli D816,测定克隆片段对pac表达的影响。如果克隆片段含有完整的调节基因(pacR)。诱导剂不能使由高拷贝pacR表达的阻抑物失活,部分阻抑物结合pac操纵基困,阻碍RNA聚合酶对加pac的转录,因此pac的表达量降低。发酵结果表明,阻抑物可能是由pac结构基因内部的ORFⅡ编码的蛋白因子。     

嗜碱芽孢杆菌XE22-4-1碱性弹性蛋白酶发酵条件的研究

从西藏天然碱湖中筛选到一株产碱性弹性蛋白酶 (alkalineelastase)菌株 ,最适生长pH为 1 0 0 ,经鉴定为Bacillussp .,编号XE2 2 4 1。该菌产酶最适碳源为 2 %葡萄糖 ,氮源为0 2 5%酵母粉 ,豆饼粉对发酵产酶有促进作用。 2L发酵罐实验表明 ,溶解氧是影响该菌株产酶的重要因素。通过提高通气量和改变搅拌速度 ,该菌株可在发酵 48h内达到产酶高峰 ,酶活力最高达 2 66u mL     

USING TREND INDICES FOR ENDANGERED SPECIES

Initial appraisals of the status of endangered large-mammal populations may have to depend on indices of population trend. Such indices may possibly be improved by using auxiliary variables. Various models were studied for populations of the Florida manatee (Trichechus manatus latirostris), Yellowstone grizzly bear (Ursus arctos horribilis), and Hawaiian monk seal (Monachus schauinslandi). Several criteria for checking validity of the fitted models were considered, and the simple R² criterion appears to provide useful comparisons. Multiple regression models overestimated the rate of change of the East Coast manatee population as determined from three other sources (a covariance model, a non-linear model, and the rate estimated from reproductive and survival data). A multiple regression model for grizzly bears using three auxiliary variables exhibited a fairly high R² (0.84) and appeared to provide a better fit than did a non-linear model. A beach count index for Hawaiian monk seals seemed to be unreliable for year-to-year comparisons in contrast to total population counts and estimates from a capture-recapture method. The use of auxiliary variables for checking and improving trend index data appears feasible and well worthwhile.     

A METHOD FOR ESTIMATING MASS OF LARGE PINNIPEDS

Fifty-two male elephant seals were weighed and photographed at Año Nuevo State Reserve, California, to establish a predictive relationship between photographically measured morphological variables (length, side area, and girth area) and body mass. Regression of mass on these variables revealed that side area, roughly equivalent to a longitudinal cross-section, was the most useful single variable for predicting mass, and that adding the other two variables to side area slightly improved the accuracy of the photogrammetric technique. Curvilinear regressions based on a power model provided the best predictive relationships. This technique may prove useful for estimating body mass of other pinnipeds.     

SENSORY OPTIMIZATION OF PROCESS VARIABLES FOR PREPARING ROASTED GRAIN POWDERS

The quality of roasted grain powders is greatly affected bythe roasting process variables; viz., initial moisture content (IMC), grain-to-sand ratio (GSR), sand temperature (ST) and roasting time (RT). The sensory optimization for these variables was done in two phases. The IMC (12 ± 0.2%) and GSR (1:4) were standardized in the first phase, whereas ST (215C) and RT (1.5–2.0 min), in the second. The overall quality of the roasted grain powders was found to be greatly affected by ST followed by RT.     

RECOGNITION MEMORY FOR ODORS

Recognition tests demonstrated unique features of odor memory.The forgetting curve over a span of one month showed a veryslow decay from an initial performance of 85% correct recognition,a finding that contrasts with the rapid decay for stimuli presentedto other sense modalities. The variables of labelling, codability,serial position, and retention interval had little or no effecton performance. Even the variables of familiarity and pleasantness,salient aspects of odor experience, had no effect on recognition.Hence, the connection between linguistic processing and odorrecognition is weak. ^* This research supported by grant ES-00592 from the NationalInstitutes of Health.     

A COMPUTERIZED SYSTEM FOR CONTROLLING AND MEASURING GUSTATORY REACTION TIMES

Reaction Time (RT) procedures are widely used in cognitive and behavioral experiments. In the sensory realm RT has been traditionally applied to measure visual, auditory or motor responses. The application of the RT method to gustatory stimuli has proved to be difficult. Attempts to develop automatic control techniques have been restrained by difficulties related to the control of variables, e.g. physiochemical characteristics of chemical solutions and the procedure for stimulus presentation. In this report we describe a computer based system that was designed to measure the reaction time to taste solutions dropped on the tongue. The equipment consists of a pumping system, an interface between the computer and the pumping system, the software required to control the interface and to measure reaction time, and a push button to detect the subject's response. The system can be used as a tool for both research and evaluation tests.     

用于全球变化研究的中国植物功能型划分

 植物功能型(Plant functional types, PFTs)作为沟通植物的结构和功能与生态系统属性的桥梁,随着全球变化与植被的关系研究的深入而受到广泛重视。植物功能型的划分依赖于研究的背景、尺度和要解决的问题。为了区域尺度全球变化研究的需要,该文提出了一个基于植物关键特征的植物功能型划分方法。该方法首先选择了6项植物特征,包括3项冠层特征：木本-草本、常绿-落叶和针叶-阔叶,以及3项生理特征：光合途径(C3 / C4)、植物的水分需求和热量需求,作为划分植物功能型的关键特征;然后,先根据植物冠层特征划分得到5个基本类型,再根据水分和热量条件进行详细划分,得到29种备选类型;需要时,再根据研究目的从这29种备选类型中选择所需类型。根据这个方法,在充分考虑了我国季风气候条件下特有的水热配置和高海拔环境对植物的形态和功能特征影响的基础上,从备选类型中选择了一套适合中国气候和植被特征的植物功能型体系。这套体系包括18类植物功能型,其中含7类‘树’功能型、6类‘灌木’功能型和5类‘草’功能型,另根据需要设置2类‘裸地’功能型。并且根据植物的生理生态特征和中国植被的地理分布确定了用于限制植物功能型分布的气候因子,这些气候因子包括绝对最低温度、最暖月平均温度、有效积温、年最热月平均温和最冷月平均温之差、湿润指数、年均降水量。应用表明,这套植物功能型可用于模拟我国植被在当前气候条件下的分布。该研究为发展适于我国的植被模型和区域气候模型、评估全球变化对我国植被的影响及植被变化对气候的反馈作用提供依据与参数。     

武汉东湖的磷-浮游植物动态模型

本文报道了东湖的一个富营养化模型。这个模型按照1年的时间标度描述东湖藻类的生长和磷循环,其状态变量包括浮游植物磷、藻类生物量、正磷酸盐、碎屑磷和沉积物磷。模型校准和检验结果表明,模型对于系统给定状态的描述是令人满意的,并且对于系统的强制函数的改变能给予合理的响应。根据东湖富营养化工程治理的初步设想,利用模型进行了东湖污水截流前后的水质预报,同时考查了截流后移出沉积物或引灌江水对于改善其水质的效果。模型所提供的各种预报可供拟定东湖治理方案时参考。     

UTILIZATION OF HAND-HELD PROGRAMMABLE CALCULATORS FOR RAPID BEDSIDE COMPUTATION OF PHYSIOLOGIC-PHARMACOLOGIC VARIABLES AND PROGNOSTIC INDICES DURING MECHANICAL CIRCULATORY SUPPORT

This study describes five programs that may be used on compact, low-cost programmable calculators with adequate memory and sufficient numbers of program steps to compute cardiorespiratory variables. These short programs are especially useful in the operating room and at the bedside.     

RELIABILITY AND VALIDITY OF AN ACCELEROMETRIC SYSTEM FOR ASSESSING VERTICAL JUMPING PERFORMANCE

The validity of an accelerometric system (Myotest©) for assessing vertical jump height, vertical force and power, leg stiffness and reactivity index was examined. 20 healthy males performed 3ד5 hops in place”, 3ד1 squat jump” and 3× “1 countermovement jump” during 2 test-retest sessions. The variables were simultaneously assessed using an accelerometer and a force platform at a frequency of 0.5 and 1 kHz, respectively. Both reliability and validity of the accelerometric system were studied. No significant differences between test and retest data were found (p < 0.05), showing a high level of reliability. Besides, moderate to high intraclass correlation coefficients (ICCs) (from 0.74 to 0.96) were obtained for all variables whereas weak to moderate ICCs (from 0.29 to 0.79) were obtained for force and power during the countermovement jump. With regards to validity, the difference between the two devices was not significant for 5 hops in place height (1.8 cm), force during squat (-1.4 N · kg⁻¹) and countermovement (0.1 N · kg⁻¹) jumps, leg stiffness (7.8 kN · m⁻¹) and reactivity index (0.4). So, the measurements of these variables with this accelerometer are valid, which is not the case for the other variables. The main causes of non-validity for velocity, power and contact time assessment are temporal biases of the takeoff and touchdown moments detection.