Ochrobactrum intermedium DN2烟碱降解酶发酵培养基优化研究

研究工作中筛选得到一株有较强烟碱降解能力的新型烟碱降解细菌 Ochrobactrum intermedium DN2,对其产烟碱降解酶的发酵培养基进行了优化.采用Plaekett-Burman设计法,对影响Ochrobactrum intermedium DN2产烟碱降解酶的15个因子进行了筛选.结果表明,影响该菌发酵产酶的显著因子为烟碱、酵母膏、葡萄糖、tween-80的质量浓度和培养基初始pH.在此基础上,采用响应曲面法对以上5个显著因子进行了优化.得到各因子最佳水平为烟碱2.183g/L,葡萄糖0.823 g/L,酵母膏0.844 g/L,Tween-80 0.976 g/L,初始pH 6.9.在此优化条件下获得实际酶活为7943 U/L,与预测值8 060 U/L相近,比优化前提高了52%.     

谷氨酸脱羧酶发酵工艺的优化

用大肠杆菌AS1.505进行液态发酵生产谷氨酸脱羧酶并优化培养基,考察了碳源、氮源、复合营养物质、起始pH及发酵时间对酶活的影响,确定最佳产酶培养基组成为:葡萄糖1.0 g/dL,蛋白胨3.0 g/dL,氯化钠0.3 g/dL,磷酸氢二钾0.1 g/dL,硫酸镁0.02 g/dL,L-谷氨酸0.01g/dL,玉米浆1.5 g/dL,生物素30 t,g/L,麸皮4 g/dL;pH 6.5.在此基础上,设计发酵条件的优化实验.实验结果表明为:250 mL的三角瓶装液量25 mL,37℃,起始pH 6.5,培养18 h达到产酶高峰,产酶活力可达1 290 U/mL.     

碳源对绿色木霉ZC产中性纤维素酶的影响

筛选得到一株高产中性纤维素酶的绿色木霉ZC,对其培养基中的碳源进行了优化,探讨了碳源的种类、混合碳源以及碳源与麸皮的比例对其产酶的影响,确定了以4 g/dL玉米秸秆粉、1g/dL麸皮为主的发酵培养基,此培养基中纤维素酶滤纸酶活可达321.12 U/mL.     

重组大肠杆菌BL21（DE3）-pET28a（＋）-bgl诱导表达β-葡聚糖酶的条件优化

研究自行构建的产β-葡聚糖酶的工程菌E.coli BL21(DE3)-pET28a(+)-bgl在LB培养基中的生长特性,考察种子液的菌龄、培养基起始pH、接种量及诱导起始时发酵液菌浓度等对β-葡聚糖酶产生水平的影响;通过正交试验确定诱导剂IPTG及乳糖添加量、诱导温度及诱导剂作用时间.结果表明:培养基起始pH 7.0,对数生长中期的种子液(OD600为0.35)以接种量(体积分数)10%接入摇瓶发酵培养,37 ℃,200 r/min培养约3 h,菌液OD值达到1.0左右,添加终浓度分别为0.033 6 mmol/L的IPTG及10 mmol/L乳糖,24℃诱导6 h,发酵液清液中酶活达到最高(336.33 U/mL),菌体生长量为1.12 g/L,发酵液中总酶活达到459.32 U/mL,是原始菌株在相同条件下所产酶活的6.62倍.采用优化培养条件及诱导剂作用条件,重组菌在TB培养基中酶活水平进一步提高,诱导剂作用10 h,发酵清液中酶活为1 090.31 U/mL,总酶活1 570.83 U/mL,是原始菌在该条件下酶活的19.73倍,显示出重组菌具有广阔的工业化应用前景.     

一株碱性低温脂肪酶产生菌发酵条件的优化

通过单因素实验,对SYBC Wu-3菌摇床发酵产脂肪酶的培养基组成和培养条件进行优化,得出较佳的产酶培养基组成配方为:蛋白胨5 g/L,酵母膏 6 g/L,NaH2PO4 3 g/L; 油脂250 mL/L,乳化剂OP 25 mL/L.最优发酵条件为250 mL的摇瓶装液量50 mL,培养温度30 ℃,发酵时间72 h.经过优化后发酵液脂肪酶酶活力最高可达到10.68 U/mL,较优化前提高了2.8倍.     

Zn2+对轮枝链霉菌SK4.001生长及转谷氨酰胺酶合成的影响

研究了Zn2+在SK4.001发酵生产转谷氨酰胺酶中的作用. 在发酵培养基中加入不同质量浓度的Zn2+,检测菌体量、pH、残余甘油及转谷氨酰胺酶(TGase) 酶活的变化, 发现Zn2+在SK4.001发酵生产转谷氨酰胺酶中除满足菌体生长需要外,还具有提高转谷氨酰胺酶酶活的作用. 发酵培养基中含有3.15 μg/mL Zn2+时,菌体生长正常,菌体量较高;当Zn2+质量浓度增加到8.15 μg/mL时,菌体量改变不明显, 但转谷氨酰胺酶酶活迅速上升,达到4.723 U/mL,是Zn2+质量浓度为3.15 μg/mL时的2.06倍.     

纳豆芽孢杆菌的固态发酵条件

为了优化纳豆芽孢杆菌固态发酵条件,以活菌数和芽孢数为指标,采用微生物发酵技术,研究了种龄、接种量、培养基初始pH值和含水量对固态发酵的影响,并通过L9(34)正交试验确定了纳豆芽孢杆菌固态发酵最佳条件.试验结果表明:接种体积分数7%、种龄17 h、培养基初始pH值8.0、培养基初始水分质量分数70%,37℃固态发酵5 d效果最好,活菌数和芽孢数分别达到3.4×1010 cfu/g和1.8×109 cfu/g.     

Brevibacterium sp.DGCDC-82发酵罐中生产胆固醇氧化酶

用Brevibacterium sp.DGCDC-82在7L发酵罐中分批培养.分别对添加剂吐温-80、培养温度、培养基的pH、通风量和搅拌速度在胆固醇氧化酶的生产中的影响进行了研究.结果显示以上条件均对产酶有影响.在不同的发酵阶段改变发酵操作条件,发酵20 h最高酶活可达1203U/L,生产强度可达60 U/(L·h).既有效地提高了胆固醇氧化酶的产量,又防止了发酵过程中的泡沫外溢.     

pH值和初始淀粉质量浓度对发酵生产谷氨酰胺转胺酶的影响

研究了不同的 pH值对谷氨酰胺转胺酶 (MTG)发酵过程中菌体生长和产酶的影响 ,在此基础上探讨了不同初始淀粉质量浓度及中后期碳源流加对发酵过程的影响 .研究结果表明 :pH值对菌体的生长和产酶模式产生明显的影响 ,当发酵过程的 pH值控制在 6 .5时 ,最有利于菌体的生长和酶的合成 ,在此条件下可得到较高的菌体干重 (DCW )和酶活 ;初始淀粉质量浓度以 3g/dL较适宜 ,其DCW和MTG酶活最高 ,DCW为 2 5.1g/L ,酶活水平达 2 .94U /mL ;中后期采用流加碳源的策略使发酵时间比分批发酵最好水平缩短 12h左右 ,酶活提高到 3.0 5U/mL ,各项指标均比分批发酵最好水平有明显的提高 .     

以玉米芯木聚糖为碳源的草菇木聚糖酶的发酵条件

在玉米芯木聚糖的碱法提取中,玉米芯木聚糖在80℃抽提120min的抽提率较高,还原糖质量分数较低,同时,抽提液的颜色较浅,明显优于121℃抽提30min.以80℃抽提的木聚糖为诱导,草菇产生的木聚糖酶的酶活比121℃抽提的木聚糖的酶活高.以80℃抽提的木聚糖为碳源,通过正交试验,设计出适合草菇生长和产生木聚糖酶的发酵条件.培养基组成为:木聚糖15g/L,(NH4)2SO42.5g/L,Tween 803g/L,酵母膏1g/L,KH2PO41g/L,MgSO4·7H2O0.6g/L,微量元素0.0005g/L.pH7.0,40℃下150r/min培养4d,此时酶活达32.4IU/mL.     

发酵甘油废水的处理工艺

针对发酵甘油生产过程中的淀粉质原料浸泡废水固体悬浮物质量浓度高和提取废水难以生物降解的特性,分别采用沉淀法和Fenton试剂法对其进行预处理,再与发酵废水混合后,采用UASB-SBR组合工艺进行处理.结果表明:当进水COD为6 800-7 360 mg／L时,处理后出水COD可降到100 mg／L以下,出水NH3-N降到15 mg／L以下,系统出水可以达标排放.     

Dioctophyme larva in the subcutaneous tissues of a woman in Ohio

A nematode larva undistinguishable from Dioctophyme renale was found in the subcutaneous tissues of the abdomen of a 23-year-old woman from Ohio. This is the third case of Dioctophyme larva reported in humans. Although it is not known how the infection was acquired, we suggest that ingestion of raw fish was the probable source. We further hypothesize that such a larva could eventually migrate to the kidney and grow to the adult state.     

精子优选后冷冻保存对活性氧生成的影响

目的观察优选技术在精液冷冻保存中对常规精液参数、线粒体膜电位(△Ψm)和活性氧(ROS)的影响。方法分别用计算机辅助精子分析仪(CASA)、流式细胞仪(FCM)检测优选或未优选的精子在冷冻前后常规精液参数、△Ψm和精子细胞内ROS的变化。结果各冻融组精子活率、a级精子、(a+b)级精子、正常形态率均比冷冻前有不同程度降低,有显著性差异;而优选冻融组和优选加精浆冻融组间常规精液参数无显著性差异(P>0.05),但都明显高于原始冻融组(P<0.01)。原始精液冷冻后比冷冻前△Ψm下降(P<0.05),ROS显著上升(P<0.01)。优选冻融组和优选加精浆冻融组与冷冻前相比,以及组间无显著性差异(P>0.05);但与原始冻融组分别比较,△Ψm均显著提高(P<0.05),ROS均显著减少(P<0.01)。各冻融组△Ψm和ROS都有明显相关性,相关系数(r)依次为-0.528、-0.537、-0.606(P<0.001)。结论精液优选后冷冻可以在保持良好的△Ψm同时,减少ROS生成,因此建议临床可应用该技术冷冻保存精子。     

不同炮制方法对黄芪化学成分的影响

目的：探讨不同炮制方法对于黄芪制品中黄酮类及糖类成分的影响。方法将统一产地及批次的黄芪通过不同的炮制方式转变为酒黄芪、米黄芪、炒黄芪、盐黄芪及蜜黄芪,分析不同方式炮制的黄芪中含有的糖类及黄酮类成分的变化情况。结果黄酮类：酒黄芪中毛蕊异黄酮的含量相对于生黄芪明显增加;蜜黄芪中黄酮类成分含量明显下降,而盐黄芪、米黄芪对黄酮类成分的影响不明显。糖类：水溶性糖在不同炮制方式中的含量由高到低依次为生黄芪＞米黄芪＞酒黄芪＞盐黄芪＞炒黄芪;还原性糖：生黄芪＞米黄芪＞酒黄芪＞盐黄芪＞炒黄芪;多糖：酒黄芪＞盐黄芪＞炒黄芪＞米黄芪＞生黄芪,以酒炙黄芪最高。结论不同的炮制方式会对黄芪中的有效成分含量产生明显的影响,黄酮类以酒制增加最为明显,蜜制则下降明显;多糖类也亦酒制增加明显,而水溶性糖及还原性糖则通过炮制后含量下降,以盐黄芪下降最明显。     

Oxidation of fatty acids is the source of increased mitochondrial reactive oxygen species production in kidney cortical tubules in early diabetes

Mitochondrial reactive oxygen species (ROS) cause kidney damage in diabetes. We investigated the source and site of ROS production by kidney cortical tubule mitochondria in streptozotocin-induced type 1 diabetes in rats. In diabetic mitochondria, the increased amounts and activities of selective fatty acid oxidation enzymes is associated with increased oxidative phosphorylation and net ROS production with fatty acid substrates (by 40% and 30%, respectively), whereas pyruvate oxidation is decreased and pyruvate-supported ROS production is unchanged. Oxidation of substrates that donate electrons at specific sites in the electron transport chain (ETC) is unchanged. The increased maximal production of ROS with fatty acid oxidation is not affected by limiting the electron flow from complex I into complex III. The maximal capacity of the ubiquinol oxidation site in complex III in generating ROS does not differ between the control and diabetic mitochondria. In conclusion, the mitochondrial ETC is neither the target nor the site of ROS production in kidney tubule mitochondria in short-term diabetes. Mitochondrial fatty acid oxidation is the source of the increased net ROS production, and the site of electron leakage is located proximal to coenzyme Q at the electron transfer flavoprotein that shuttles electrons from acyl-CoA dehydrogenases to coenzyme Q.     

Shortage of perioperative drugs: implications for anesthesia practice and patient safety

Several medications used in clinical perioperative medicine are currently cited on the national shortage list. Medication shortages may be attributed to lack of raw materials, manufacturing issues, and discontinuation of production. Medication shortage has a substantial impact on patient care, and is responsible for creating an environment conducive to an increase in medication errors. Anesthesiologists should be taking an active role with the pharmacy and hospital management to alert caregivers and help to prevent adverse effects on patient care and safety.     

Prostate epithelial cells utilize glucose and aspartate as the carbon sources for net citrate production

The prostate gland (human, rat ventral prostate) has the major function of accumulating and secreting extremely high levels of citrate. This function requires unique and specialized metabolic pathways associated with prostate secretory epithelial cells by which exogenous substrates must be utilized as the six-carbon sources of citrate. Recent studies demonstrated that aspartate can serve as the four-carbon source of oxalacetate for citrate synthesis. Identification of the two-carbon source of acetyl CoA (AcCoA) had not been established. The present study investigated the probability that exogenous glucose, via pyruvate oxidation, is a physiological source of AcCoA for net citrate production by isolated epithelial cells from rat ventral prostate. Under adequate oxygenation, 5 mM glucose in the presence of aspartate plus glutamate markedly stimulated citrate production. Exogenous and endogenous pyruvate also stimulated net citrate production. We propose that glucose via aerobic glycolysis and pyruvate oxidation provides AcCoA, which condenses with oxalacetate obtained from aspartate transamination for citrate synthesis. Prostate epithelial cells do not readily oxidize citrate, which permits accumulation and secretion of the synthesized citrate.     

Colonic healing: a role for polymorphonuclear leucocytes and oxygen radical production

Using a model of 'reperfusion injury' as a mechanism of oxygen radical (OR) production in vivo, we have demonstrated that superoxide (O2-) is the major source of damage. Inhibition of its production from polymorphonuclear leucocytes (PMNs) by aprotinin, a protease inhibitor, or by scavenging with superoxide dismutase (SOD), resulted in reduced oxidation of of tissue glutathione (P less than 0.002). Colonic healing after anastomosis was also improved by aprotinin as measured by histology (P less than 0.05), breaking energy (P less than 0.001), breaking strength (P = 0.02), and by hydroxyproline content (P = 0.01). Allopurinol, although inhibiting glutathione oxidation, had no effect on PMN leucocyte OR production and did not protect against histological damage. These data demonstrate that PMNs endogenous to the tissue or attracted there as a result of trauma are the source of OR production in 'reperfusion'. Their inhibition improves colonic healing.     

双层探测器光谱CT在心脏成像中的应用进展

双层探测器光谱CT(SDCT)具有上下两层空间对等的探测器结构,使高、低能2套数据在原始数据域空间即被解析,可用于重建反映康普顿效应和光电效应不同组成的能谱图像,并可合并重建完全等同于普通CT所具有的常规多能图像。SDCT的能谱数据具有同时、同源、同向特征,提升了其在心脏成像中的应用价值。本文对SDCT在改善冠状动脉CTA图像质量、“双低”成像、显示斑块及支架以及心肌灌注方面的应用价值进行综述。