响应面法优化北五味子褐斑病内生生防真菌淡紫拟青霉WG9发酵工艺及发酵产物稳定性研究

为提高北五味子内生生防真菌WG9发酵液对褐斑病病原菌的抑菌率,本研究采用单因素实验及响应面法对其发酵条件进行优化.首先采用单因素实验,以发酵时间(h)、发酵温度(℃)、摇床转速(r/min)及初始pH为考察指标,测定对病原菌的抑菌率及生防真菌WG9的菌丝体生物量,以获取单因素实验的最优值,然后使用Box-Behnken...     

2-氧代-4-苯基丁酸乙酯还原酶产生菌筛选及产酶条件

研究了利用生物催化不对称还原的方法制备(R)-2-羟基-4-苯基丁酸乙酯[(R)-HPBE]。以2-氧代-4-苯基丁酸乙酯(OPBE)为底物,通过对实验室保藏菌株进行筛选,得到一株产物立体选择性较高的菌株G2ndida krusei SW2026,并对其发酵产酶条件进行研究。其最适的发酵培养基组成为4.5%葡萄糖,3%蛋白胨,1.5%牛肉膏,0.05%Mn~(2+);适宜的产酶发酵条件为初始pH 6.0,温度28℃,摇床转速180 r/min,发酵周期48 h。将此条件下发酵培养的菌体用于OPBE的不对称还原反应,产物(R)-HPBE的对映体过量值(e.e.)可达97.33%,产率最高达到72.54%。     

产低温脂肪酶菌株CZW001发酵培养基优化研究

【目的】研究产低温脂肪酶菌株CZW001发酵培养基。【方法】在单因素试验的基础上, 采用Plackett-Burman (P-B)设计, Box-Behnken (B-B)设计和响应面试验设计(RSM), 在20 °C、pH 8.0、160?r/min发酵2 d条件下, 对发酵培养基进行优化。【结果】该菌株最适产酶培养基为(g/L): 葡萄糖7.68, 橄榄油21.93, 硫酸铵2.0, 磷酸二氢钾1.0, 硫酸镁0.27, 氯化钙0.3, 氯化钠20.0, 吐温-80 1.0。其最高酶活为62.8 U/mL, 比优化前提高了3.14倍。【结论】通过对产低温脂肪酶菌株CZW001发酵培养基优化研究, 明显提高低温脂肪酶活力。     

生物破乳菌的筛选及发酵条件的优化

生物破乳剂的开发可以降低油田乳状液对石油工业和生态环境的负面影响并减少化学破乳剂的使用量。本研究建立了一套高效、便捷的破乳菌筛选方法, 并对破乳菌的特性进行研究。利用大庆油田受石油污染土壤作为菌种来源, 将Tween 60-water(0.072%, V/V)和Span 60-oil (0.028%, V/V)以6.5:3.5体积比配置出可以稳定200 h以上的O/W型乳状液, 用于破乳菌破乳效能的评价。经过分离纯化、血平板试验、排油试验和破乳试验最终筛选出2株24 h平均排油率在80%以上的优势破乳菌, 初步鉴定为芽孢杆菌属(Bascillus)。通过该破乳菌发酵条的件优化得到, 当温度为25°C, 摇床转数为160 r/min, pH值为9, 接菌量为20%时对该破乳菌生长速率最快, 积累发酵产物的量最多; 当温度为35°C, 摇床转数为120 r/min, pH值为9, 接菌量为2%时该破乳菌代谢产物的破乳活性最高。     

Rates of ammonia release from sediments by chironomid larvae

SUMMARY. 1. Microcosms of Lake Balaton mud and sterilized sand and aerated water were used to evaluate ammonia increments in the overlying water as influenced by chironomid density and temperature. In the two approaches, the effects of sediment disturbance and metabolic excretion of chironomids were measured.
2. The activity of larvae increased the ammonia content of the overlying water at temperatures above 10°C. A rise of temperature to 20°C resulted in a 5–20-fold increase in ammonia release in both systems with chironomids.
3. At 10°C combined effects of sediment disturbance and of excretion produced lower release rates than did excretion rates alone (mud-water v. sand-water treatments). At higher temperatures (15 and 20°C) release rates of ammonia by sediment disturbance plus excretion were higher than excretion rates alone. Ammonia excretion contributed significantly to the total release at each temperature.
4. Metabolic mineralization of nitrogen compounds appears to be an important mechanism contributing to nitrogen regeneration from aerobic lake sediments. High N:P ratio (14:1) of chironomid excretion materials supports this interpretation.     

How does the vertical distribution of chlorophyll vary in littoral sediments of small lakes?

1. In this study, sediment chlorophyll profiles at twenty littoral stations in three oligo‐mesotrophic lakes were compared to test whether the vertical distribution of chlorophyll is related to site characteristics (light availability, temperature, physical disturbances) and whether these profiles differ between shallow and deep portions of the littoral zone.
2. The magnitude of chlorophyll peaks at the sediment surface did not vary with light intensity. Chlorophyll peaks in the shallow littoral zone had a weak tendency to decrease with increasing effective fetch. The magnitude of chlorophyll peaks at deeper sites was more closely related to water temperature than to substrate slope.
3. High chlorophyll concentrations were measured down to 1–3 cm in the sediments, both at shallow (< 2.5 m) and deep (4–10 m) stations. The depth to which high chlorophyll was found in sediments did not vary with effective fetch or sediment water content, two indices of wave disturbance in the shallow littoral zone, or with substrate slope, an index of sediment stability in the deep littoral zone. Sediment mixing is apparently not related to common indices of physical disturbances.
4. Between 8 and 100% of sediment surface chlorophyll was 'retained' 4–5 cm into the sediments. The proportion of chlorophyll 'retained' in littoral sediments increased with increasing depth, increasing lake productivity (total phosphorus concentration) and increasing lake pH.
5. Among‐core variability (standard error/mean) in chlorophyll concentration at the sediment surface ranged from less than 1% to 33% at different stations and was highest at shallow, exposed sites. These levels of variability are similar to those found in other periphytic communities.     

Carboxylmethylation affects the proteolysis of myelin basic protein by Staphylococcus aureus V8 proteinase.

Bovine myelin basic protein (MBP), charge isoform 1 (C1) was carboxylmethylated by the enzyme D-aspartyl/L-isoaspartyl protein methyltransferase (EC. 2.1.1.77) and the carboxylmethylated protein was subjected to proteolysis by sequencing grade staphylococcal V8 proteinase at pH 4.0 to identify its carboxylmethylated modified aspartate and/or asparagine residues which are recognized by this methyltransferase. Native MBP, C1 was treated similarly and the proteolysis products were compared, using electrophoretic, chromatographic and amino acid sequencing techniques. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed differences in the kinetics of proteolysis between the native and the carboxylmethylated MBP, C1 which were confirmed using HPLC. Partial sequencing of the native and carboxylmethylated fragments eluting at about 29 min (P29) revealed cleavage of native MBP, C1 at Gly-127-Gly-128 and of the carboxylmethylated MBP, C1 at Phe-124-Gly-125. Additional evidence including tryptic subdigestion of carboxylmethylated P29 disclosed the following partial sequence for this peptide: Gly-Tyr-Gly-Gly-Arg-Ala-Ser-Asp-Tyr-Lys-Ser-Ala-His-Lys-Gly-Leu-Lys- Gly-His-Asp-Ala-Gln-Gly-Thr-Leu-Ser-Lys-Ileu-Phe-Lys-. This sequence matches MBP residues 125-154. As a result of these findings, Asp-132 and Asp-144 were identified as two of the modified (isomerized or racemized) methyl-accepting L-aspartates in MBP. The results of the proteolysis experiments wherein the sequencing grade staphylococcal V8 proteinase was used at the rarely tested pH of 4.0, rather than at its commonly tested pH of 7.8, also disclose that the proteinase totally failed to recognize and hence cleave the two Glu-X bonds (Glu-82-Asn-83 and Glu-118-Gly-119) of MBP, preferring to cleave the protein at a number of hitherto unreported sites.     

Concentrations and distribution of Fe,Zn and Cu in tissues of the white sucker (Catostomus commersoni) in relation to elevated levels of metals and low pH

This study examines whether the process of lake acidification influences the accumulation of Fe, Zn and Cu in the tissues of the white sucker (Catostomus commersoni). Concentrations of Fe, Zn and Cu were measured in the liver, kidney and muscle of white sucker sampled from 4 acidic (pH range 4.8–5.3), 1 slightly acidic (pH = 5.8) and 3 circumneutral (pH = 6.3, 6.4) lakes located in south-central Ontario, Canada. Pearson product-moment correlation coefficients were used to determine relationships between average elemental concentrations in the 3 tissues and both sediment and water metal concentrations plus lake pH, DOC and alkalinity. Despite the 1000-fold difference in H+ concentration among the 8 study lakes, tissue concentrations of Fe, Zn and Cu did not correlate with lake pH. Average Fe, Zn and Cu tissue concentrations did not correlate with metal concentrations in lake water. Only Zn concentrations in the liver and muscle were correlated with Zn concentrations in the sediment (r = 0.83 and r = 0.88, P < 0.05). Iron and Cu were regulated by the white sucker over a wide range of lake pH and metal concentrations in both the water and sediment. In contrast, Zn tissue concentrations were correlated with sediment Zn concentrations, the latter are thought to result from Zn inputs of anthropogenic origin.     

地衣芽孢杆菌D-1产碱性蛋白酶培养条件的优化

为了对产碱性蛋白酶的地衣芽孢杆菌D-1的培养条件进行优化,利用10 L发酵罐,采用正交设计19(34)试验,对培养温度、pH值、搅拌转速、通气量4条件进行优化,得到地衣芽孢杆菌D-1发酵产碱性蛋白酶的最优培养条件为:培养温度37.0℃,pH值7.5,通气量4L/min,搅拌转速300r/min.利用最优条件组合进行验证...     

纳豆激酶液态发酵条件的研究

考察纳豆杆菌在7.5 L发酵罐中分批发酵产纳豆激酶的条件,纳豆激酶酶活采用四肽底物测定。结果表明:纳豆杆菌生长和产酶的适宜条件不一致。发酵过程中发酵罐搅拌转速控制为500 r/min不变,0～12 h时控制pH为8.0、温度为37℃;12～36 h调整pH为7.0、温度为30℃;16 h时补加外源C源,连续发酵36 h。该过程中发酵液中比酶活最高达到3 232 U/mL,与摇瓶发酵相比,比酶活提高了58%。     

Climate drivers of diatom distribution in shallow subarctic lakes

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一株病原拮抗野生菌株的筛选、鉴定及其发酵工艺优化

为获得广谱抗菌功能野生菌株并提高其发酵产物中抗菌物质的含量。采用管碟法和菌丝生长速率法筛选功能菌株,ITS序列分析鉴定功能菌株,通过响应面法和正交设计优化发酵生产抗菌物质的工艺。筛选到一株强效、广谱抗菌功能菌株,鉴定为Cerrena sp.,其发酵产物对金黄色葡萄球菌、大肠杆菌、白色念珠菌、枯草芽孢杆菌和水稻纹枯病菌有显著拮抗作用。该菌株的摇甁发酵配方及培养条件为：马铃薯13.99 g/L,蔗糖 41.58 g/L,VB1 0.027 g/L,麸皮7 g/L,KH₂PO₄ 2 g/L,MgSO₄·7H₂O 2 g/L;摇床温度28 ℃、发酵周期10 d、种龄4 d、接种量8%、初始pH为5.0、装液量110 ml/250 ml。该菌株有明显抑菌活性,发酵工艺优化后抗菌活性提高了30.37%,为该菌株今后的应用、抗菌剂的分离提纯和产业化提供了实验依据。     

高产洛伐他汀棒曲霉菌株的筛选、鉴定及发酵条件优化

从不同生境(食品、土壤、空气、有机质等)收集到的自然发酵样品中分离得到150株曲霉属菌株.用高效液相色谱(HPLC)法检测发酵液中洛伐他汀(Lovastatin)含量,筛选获得1株稳定高产Lovastatin的曲霉菌株(编号:Ac-32).根据菌落形态特征并结合18 S rDNA测序,鉴定其为棒曲霉(Aspergillus clavatus).通过摇瓶发酵单因素实验优化了碳氮源种类、碳氮源含量、碳氮比(C/N)、发酵温度、初始pH、转速、种龄和接种量,确定了棒曲霉菌株Ac-32摇瓶发酵产Lovastatin的适宜条件为:乳糖为碳源、蛋白胨为氮源、碳源含量为100 g/L、氮源含量为12 g/L、碳氮比(C/N)为15:1.8、温度28℃、转速180 r/min、初始pH 5.2、种龄4 d、接种量6%.采用Minitab 17软件的P-B实验设计法,筛选对Lovastatin产量有显著影响的因素为:温度、pH、碳源含量和氮源含量.根据P-B实验结果,运用响应面法分析,确定棒曲霉菌株Ac-32产Lovastatin的最优条件为:碳源含量100 g/L,氮源含量11.8 g/L,温度28℃,pH 5.2.在此条件下,Lovastatin最高产量为236.221μg/mL.     

响应面法优化灰色链霉菌产纤维素酶的工艺研究

以灰色链霉菌为原料,在单因素试验的基础上,采用响应面法试验,优化灰色链霉菌产纤维素酶活性的发酵条件。结果表明,单因素试验灰色链霉菌产纤维素酶活性的最适发酵条件:碳源为CMC-Na,氮源为明胶,温度为28℃,pH为7.0,转速为130 r/min。响应面法试验优化灰色链霉菌产纤维素酶活性最佳发酵条件为:温度27.7℃,pH值6.9,转数130.3 r/min,在此优化条件下,灰色链霉菌产纤维素平均酶活性为6.103 U/mL(n=3),与模型的预测值(6.217 U/mL)比较接近,误差为1.83%,证明了该响应面模型具有可靠性。     

Determinants of pollen dispersal in the East Asian steppe at different spatial scales

Two sites from the East Asian steppe, the Mu Us Sandland as a regional case and the Anguli Nuur catchment as a local one, were chosen to detect roles of vegetation, climate, landform, and human disturbance on pollen dispersal. 1) Vegetation: The semi-arid steppe vegetation is characterized by Artemisia and Chenopodiaceae pollen under various vegetation conditions; however, no evident correlation between pollen percentages and corresponding plant species cover is found. 2) Climate: Samples under different Mean Annual Precipitations (MAPs) clearly distinguish themselves in the Mu Us Sandland, implying MAP-determined surface pollen spectra in regional scale. 3) Landform: Surface pollen assemblages in the azonal psammophytic vegetation and lowland meadow, show insignificant variance from the zonal steppe vegetation. The azonal halophilous vegetation, mostly distributed in the lowland or near the lake shore, leads to higher than average percentages of Chenopodiaceae pollen. Signal of exotic Pinus pollen is also strengthened in the lowland and lake sediment. 4) Human disturbance: The role of human disturbance on surface pollen assemblages is weak, as shown in the Mu Us Sandland.

This study also provides theoretical bases for quantitative reconstructions of palaeoclimate and palaeovegetation based on fossil pollen spectra from lake sediments and lowland soils in the semi-arid East Asian steppe. We suggest that calibration against locally dispersed pollen taxa is necessary to reliably reconstruct changes in vegetation pattern through time, for example, a factor of 1.75 for the widely used A/C (Artemisia/Chenopodiaceae) ratio is suggested according to the pollen assemblages in the surface layers of the lake sediment vs. slope soil in Anguli Nuur. However, uncertainties also exist for such calibration considering the dynamics of local-scale azonal vegetation.     

Effects of parabolic flight on abdominal arterial pressure in conscious rats.

Abdominal arterial pressure during parabolic flight was measured using a telemetry system to clarify the acute effect of microgravity on hemodynamics in conscious rats. The microgravity condition was elicited by three different levels of entry gravity, i.e. 2 G, 1.5 G and 1 G. On exposure to 2 G, mean aortic pressure (MBP) increased up to 118.7 mm Hg +/- 7.3 compared with the value at 1 G (107.0 +/- 6.3 mm Hg, n=6). The value at microgravity preceded by 2 G was 118.0 mmHg +/- 5.2 mm HG and it was still higher than at 1 G. When 1.5 G was elicited before microgravity exposure, MBP also increased (1.5 G: 114.9 +/- 5.3 vs 1 G: 105.8+/-5.0 mm Hg) and the value at microgravity was 117.3 + /- 5.3 mmHg. During pre-microgravity maneuver with 1 G, no changes were observed compared with the control level at 1 G (pre-microgravity: 105.0 +/- 5.0 vs 1G: 104.8 +/- 5.1 mm Hg ), whereas the MBP increased up to 117.0 +/- 6.5 mm Hg on exposure to microgravity. From these results, we found that in conscious rat MBP increase during acute microgravity exposure with either 1 G or hyper-G entry.     

Changes in regional cerebral blood flow distribution during postexercise hypotension in humans.

This investigation compared patterns of regional cerebral blood flow (rCBF) during exercise recovery both with and without postexercise hypotension (PEH). Eight subjects were studied on 3 days with randomly assigned conditions: 1) after 30 min of rest; 2) after 30 min of moderate exercise (M-Ex) at 60-70% heart rate (HR) reserve during PEH; and 3) after 30 min of light exercise (L-Ex) at 20% HR reserve with no PEH. Data were collected for HR, mean blood pressure (MBP), and ratings of perceived exertion and relaxation, and rCBF was assessed by use of single-photon-emission computed tomography. With the use of ANOVA across conditions, there were differences (P < 0.05; mean +/- SD) from rest during exercise recovery from M-Ex (HR = +12 +/- 3 beats/min; MBP = -9 +/- 2 mmHg), but not from L-Ex (HR = +2 +/- 2 beats/min; MBP = -2 +/- 2 mmHg). After M-Ex, there were decreases (P < 0.05) for the anterior cingulate (-6.7 +/- 2%), right and left inferior thalamus (-10 +/- 3%), right inferior insula (-13 +/- 3%), and left inferior anterior insula (-8 +/- 3%), not observed after L-Ex. There were rCBF decreases for leg sensorimotor regions after both M-Ex (-15 +/- 4%) and L-Ex (-12 +/- 3%) and for the left superior anterior insula (-7 +/- 3% and -6 +/- 3%), respectively. Data show that there are rCBF reductions within specific regions of the insular cortex and anterior cingulate cortex coupled with a postexercise hypotensive response after M-Ex. Findings suggest that these cerebral cortical regions, previously implicated in cardiovascular regulation during exercise, may also be involved in PEH.     

响应面法对微拟球藻产微藻蛋白的发酵条件优化

以稳定期微藻蛋白浓度为评价指标,利用响应面设计对微拟球藻(Nannochloropsis gaditana)的分批发酵条件进行优化。在单因素试验的基础上,选取温度、p H、搅拌速度及通气量为影响因子,采用四因素三水平的Box-Benhnken中心组合法设计试验。结果表明:微拟球藻的最佳发酵条件为温度30℃、p H 6.9、搅拌速度340 r/min以及通气量0.65 vvm,在此优化条件下得到微藻蛋白浓度为6.18 g/L,与模型预测值基本相符,较优化前提高了9.18%。     

乌勇布拉克盐湖嗜盐古菌絮凝效果筛选及活性检测

【目的】从嗜盐古菌中筛选可产生生物絮凝剂的菌株,对发酵液、上清液、菌悬液、胞外聚合物的絮凝作用进行检测,筛选能够适应高盐废水处理,且具有广谱盐度及pH作用范围的微生物絮凝剂。【方法】以新疆乌勇布拉克干盐湖沉积物为研究对象,利用纯培养方法对嗜盐古菌进行分离,对絮凝菌株进行初筛及16S rRNA基因测序,构建系统进化树,初步判断菌株分类地位;复筛检测不同生物材料的絮凝效果;选择絮凝效果较好的生物材料,检测其盐度、pH的絮凝效果稳定性。【结果】采用纯培养方法共分离到28株嗜盐古菌,絮凝初筛共筛选出16株嗜盐古菌,分布于碱线菌属(Natrinema)、盐缓长菌属(Halopiger)和盐土生菌属(Haloterrigena)。菌株发酵液、上清液、菌悬液、胞外聚合物具有不同程度的絮凝效果。菌株A279-1、A133、RP33、NGA0064、RM-152、A389的发酵液、上清液的絮凝效果较好,其中菌株A389的发酵液絮凝率为61.06%,上清液为67.92%。所有菌株菌悬液的絮凝率达到80%以上。菌株所产胞外聚合物表现出较好的絮凝效果,菌株RM-152所产胞外聚合物的絮凝率最高,达89.86%,其次是A389 (81.53%)。菌株A389所产胞外聚合物的产量最大,达12.53 g/L,具有广泛的盐度和pH适应性。【结论】乌勇布拉克干盐湖沉积物中蕴含丰富的可产生微生物絮凝剂的嗜盐古菌资源。嗜盐古菌菌株发酵液、上清液、菌悬液及胞外聚合物均具有良好的絮凝作用,尤其是胞外聚合物表现出较好的絮凝效果,具有广谱的盐度和pH耐受性。嗜盐古菌所产生物絮凝剂的发现对于后续高盐废水功能材料开发具有重要应用价值。     

Sedimentary microbial community dynamics in a regulated stream: East Fork of the Little Miami River,Ohio

A field study was conducted in the Lower East Fork of the Little Miami River, a regulated stream in Clermont county, Ohio, to determine how changes in streamflow, water temperature and photo-period affect sediment microbial community structure. Surface sediment cores were collected from sampling stations spanning 60 river kilometers three to four times per year between October 1996 and October 1999. During the final year of the field study, water temperature, water depth, conductivity, total suspended solids, dissolved organic carbon, instantaneous streamflow velocity, sediment grain size and sediment organic matter were determined. Total microbial biomass was measured using the phospholipid phosphate technique (PLP) and ranged between 2 and 134 nmol PLP * g(-1) dry weight sediment with a mean of 25 nmol PLP * g(-1). Microbial community structure was determined using the phospholipid fatty acid analysis and indicated seasonal shifts in sedimentary microbial community composition. January to June sedimentary microbial biomass was predominately prokaryotic (60% +/- 2), whereas microeukaryotes dominated samples collected during the late summer (55% +/- 2.4) and fall (60% +/- 2). These changes were correlated with stream discharge and water temperature. Microbial community structure varied spatially about a reservoir with prokaryotic biomass dominant at upstream stations and eukaryotic biomass dominant at downstream stations. These findings reveal that sedimentary microbial communities in streams are dynamic responding to the seasonal variation of environmental factors.