PBD和RSM联用优化聚酯型儿茶素A化学合成技术参数

为探明化学合成条件对聚酯型儿茶素A（Theasinensin A,TSA）得率的影响,通过单因素试验和Plackett-Burman Design（PBD）确定TSA化学合成关键因子,然后采用响应面法（Response surface methodology,RSM）,进一步优化TSA化学合成技术参数。结果表明,氯化铜用量、甲醇体积分数、温度对TSA得率的影响差异极显著,主因素效应为甲醇体积分数>氯化铜用量>温度,最优条件为氯化铜用量43%,甲醇体积分数26%,温度15℃,聚酯型儿茶素得率为59.12%,与模型预测值59.34%接近。PBD和RSM联用优化TSA化学合成工艺可行,预测性较好,可为其他种类儿茶素氧化聚合物的高效化学合成提供借鉴和理论依据。     

Factors affecting the virulence of Ralstonia solanacearum and its colonization on tobacco roots

Tobacco bacterial wilt caused by Ralstonia solanacearum is a serious disease affecting tobacco cultivation in southwest China. The response surface methodology was employed to evaluate the optimal conditions of tobacco bacterial wilt, and green fluorescent protein gene (gfp) labelling was applied to monitor the location and survival dynamics of R. solanacearum (Rs::gfp) on tobacco roots and in soil under these optimal conditions. The results showed that the highest wilt incidence was 91.13%, which occurred when the population reached 6.6 × 10⁶ CFU/g soil, the temperature was 30.55 °C, and the humidity was >81.42%. The Rs::gfp densely colonized the root tips and root hairs, and cells of Rs::gfp were observed intermittently in the elongation zone or at the point of the emerging lateral roots. The Rs::gfp number in the rhizosphere soil was 10.75‐, 73.13‐ and 74.86‐times higher than that in the bulk soil at 10, 15 and 20 days after transplantation, respectively. Increased colonization by Rs::gfp was related to the population of the pathogen, the environmental temperature and the humidity in the soil. These three conditions determined whether R. solanacearum would induce tobacco wilt. This is the first study to investigate factors affecting the virulence of a tobacco wilt bacterial pathogen, which is important for conducting field diagnosis and biocontrol of tobacco bacterial wilt.     

Optimization of d-pinitol extraction from vegetable soybean leaves and its potential application in control of cucumber powdery mildew

d-pinitol is a bioactive compound with an important application in both food and nonfood industries. In the present study, the leaves of twelve vegetable soybean cultivars that are usually treated as agricultural waste were screened for the presence of d-pinitol. The results showed that the d-pinitol content in these 12 cultivars ranged from 1.32 to 3.04% (w/w) of dry weight, and the cultivar Z98002 was found to contain the highest amount of d-pinitol. Response surface methodology (RSM) was used for optimization of d-pinitol extraction conditions by the Box–Behnken design. The predicted extraction efficiency, which is at least 1.3-fold that of the previously reported typical method, reached to 3.27% under the following optimal conditions: temperature of 65.5 °C, extraction time of 86.8 min, and dilution rate of 1:10. The test in the growth chamber showed that the efficacy of soybean extracts in cucumber powdery mildew control was dose-dependent on the d-pinitol concentration, and the formulated extracts clearly increased the control compared to the crude extract. This effect of the d-pinitol formulation in a greenhouse was confirmed in four provinces of China over a period of two years. Our present study provides the optimized extraction conditions for a selected agricultural waste—vegetable soybean—to obtain a high yield of d-pinitol. Results from the growth chamber test and the greenhouse bioassay will be useful in exploring a low-cost phytochemical fungicide for cucumber powdery mildew control.     

海洋芽孢杆菌N11-8产蛋白酶的发酵条件优化

本研究采用响应面法(RSM)对南极海洋芽孢杆菌(Bacillus sp.)N11-8液体发酵产蛋白酶PBN11-8的发酵条件进行快速优化。通过单因素实验初步确定南极海洋芽孢杆菌N11-8产蛋白酶的最佳碳源和氮源分别为可溶性淀粉和蛋白胨；并通过Plackett-Burman(PB)设计对影响其产酶相关因素进行评估，筛选出具有显著效应的3个因素：温度、氮源和碳源；以最陡爬坡实验逼近至上述因子最大响应区域，进而采用RSM法对其最佳水平范围进行研究，确定最优发酵条件为：可溶性淀粉3 g/L，蛋白胨13.1 g/L，酵母浸粉2.9 g/L，NaCl 5 g/L，KH2PO4 1 g/L，FeCl3·6H2O 2 mmol/L，初始pH值为7.0，温度为34.0℃，转速为200 r/min，装液量为50 ml/250 ml，接种量为4%，培养时间为60 h。最终优化后的酶活达到90.5 U/ml，比初始酶活提高了23.6%。     

响应面优化甘草浸膏中甘草酸超声提取工艺

[目的]利用响应面法优化超声提取甘草浸膏中甘草酸的工艺条件。[方法]在乙醇浓度、超声波时间及料液比等单因素试验的基础上,根据Box-Behnken的中心组合设计原理采用3因素3水平的响应面分析法优化超声波提取甘草酸的工艺条件。[结果]超声提取甘草酸的最佳工艺条件为乙醇浓度70%、超声波时间30 min、料液比2.8 g/L,在此条件下甘草酸含量为8.34%。[结论]建立了甘草浸膏中甘草酸超声提取最佳工艺条件,为甘草浸膏的精深加工及进一步研究甘草酸在食品和医药领域产业化应用提供理论依据。     

烟草青枯病菌在烟草根际的定殖及最适发病条件

为明确烟草青枯病菌在根际的定殖情况和最适发病条件,采用绿色荧光蛋白(gfp)标记病原菌,监测其在根际的定殖位置及数量动态变化,并运用荧光定量PCR法测定病原菌在根际和土体土壤的数量,通过响应曲面法探索病原菌致病的临界浓度、发病最适温湿度以及对烟株青枯病发病的影响。结果表明,从贵州省烟田土壤中分离到烟草青枯病病原菌茄科劳尔氏菌Ralstonia solanacearum,gfp-标记的茄科尔劳尔氏菌主要定殖于根尖和根毛区,少数菌落定殖于初生根的延长区,定殖部分表现为非连续性;第10、15、20天根际土壤的茄科劳尔氏菌数量均显著高于土体土壤数量,分别是土体土壤中的1.15、1.33、1.42倍;当土壤中每克土病原菌数量为106.82 CFU、温度为30.55℃、相对湿度为81.42%以上时,烟草青枯病的病情指数最高为91.13,烟株易发生青枯病。表明茄科尔劳尔氏菌在根际土壤中比土体土壤更易定殖,烟草青枯病发病条件主要取决于病原菌浓度、温度以及相对湿度3项指标。     

蜂蜜火龙果花茶饮料的工艺优化

为有效利用火龙果花,提高火龙果农户收入,以大红龙品种火龙果花以及脱水火龙果花浸提液为主要原料,研制满足大众口味的蜂蜜火龙果花茶饮料。通过单因素试验探讨浸出液、蜂蜜、冰糖、柠檬酸、纯水、花干质量以及均质时间对蜂蜜火龙果花茶饮料感官评价分数的影响。在单因素试验的基础上采用响应面分析法(RSM)对浸出液、蜂蜜、柠檬酸比例进行优化。结果表明:蜂蜜火龙果花茶饮料的最佳配比为浸出液体积342.15mL,蜂蜜质量79.73g,柠檬酸质量1.62g,冰糖质量25g,纯水体积450mL,火龙果鲜花质量60g,均质时间0min,在此条件下蜂蜜火龙果花茶饮料的感官评价分数为84.79。制得饮料酸甜爽口、风味独特、生津止渴,具有较好的应用前景。     

基于响应面分析方法的氧化乐果降解条件优化

[目的]采用响应曲面法对影响氧化乐果降解的因素进行优化。[方法]选取5个因素pH值（A）、接种量（B）、装液量（C）、温度（D）和摇床转速（E）的最佳水平范围进行研究。通过对二次多项回归方程求解获得氧化乐果降解的最佳条件。[结果]氧化乐果降解的优化条件为：pH值7.10,接种量9.46%,装液量86.38ml,培养温度30.25℃和摇床转速165.32r/min。在此最优条件下,氧化乐果的降解率最大,响应预测值为71.78%,验证值为71.40%。[结论]该研究为降低氧化乐果对动物体的危害提供科学依据。     

黑曲霉固体发酵产木聚糖酶的响应面优化设计及其酶学性质的研究

本研究采用响应面法的中心组合对影响黑曲霉(aspergillus niger JL 15)固体发酵产木聚糖酶的条件进行了优化.以橘皮粉为基质,补充碳源、氮源,以及含水量和发酵时间对木聚糖酶产量具有显著影响(P<0.05).模拟二次多项式回归预测模型,并建立自变量与响应值的回归方程,获得各因素的最佳水平,即:甘油、硫酸铵的添加量分别为4.2%和3.1%,含水量为61%,发酵时间为73.4 h,木聚糖酶活性最大预测值达922.9 U/g干发酵产物,验证值为917.7 U/g干发酵产物,高出基础培养基酶活3.2倍.酶学性质分析研究表明,黑曲霉木聚糖酶(XylA)的最适温度和最适pH分别为55oC和pH 5.0.XylA的K_m和V_(max)值分别为9.24 mg/mL和54.05 μmol/min/mL.Mn~(2+)、Zn~(2+)和斗和Mg~(2+)对XylA活性具有促进作用,而Fe~(3+)和cu~(2+)对XylA活性具有明显抑制作用.HPLC分析结果表明,XylA的水解桦木木聚糖和麸皮不溶性木聚糖的产物均为木糖至木六糖,主要产物为木三糖.     

秸秆可溶性糖测定时样品前处理条件的优化

为确定秸秆可溶性糖测定时样品的最佳前处理条件,研究单因素对玉米秸秆和小麦秸秆可溶性糖得率的影响。利用中心组合试验(Box-Behnken)和响应曲面法(RSM)对主要前处理参数进行优化,并得到回归模型。结果表明:固液比(秸秆质量与水体积比)、水浴温度、提取时间和存放时间对玉米秸秆和小麦秸秆可溶性糖得率影响显著(P<0.05)。回归方程较好的反映了可溶性糖得率与固液比、水浴温度和提取时间的关系。玉米秸秆可溶性糖测定的最佳前处理条件为,固液比0.31g/mL,水浴温度94℃,提取时间63min,此条件下的可溶性糖得率为3.85%。小麦秸秆可溶性糖测定的最佳前处理条件为,固液比0.33g/mL,水浴温度95℃,提取时间58min,此条件下的可溶性糖得率为3.39%。