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.     

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.     

Thermobifida fusca WSH03-11突变株合成角质酶的发酵条件

以放线菌Thermobifida fusca WSH03—11——高产角质酶突变株为出发菌株，在摇瓶中考察了碳、氮源等条件以及接种量、装液量和初始pH值等环境条件对突变株细胞生长和产角质酶的影响。通过优化发酵条件，突变菌株的角质酶酶活达10．1U／mL，提高了15％。在此基础上，在5L发酵罐中进一步研究了碳源流加对突变株发酵产酶的影响，结果发现，分别在0、24、48h添加1％乙醇，角质酶酶活达到16．4U／mL，细胞干重（DCW）达到3．7g／L，而且发酵时间也由110h缩短到50h。     

Kluyveromyces fragilis LFS-8611 β-D-半乳糖苷酶的摇瓶培养条件

脆壁克鲁维酵母(Kluyveromycesfragilis)LFS 8611合成的β D 半乳糖苷酶具有较高的催化半乳糖基转移反应活力.脆壁克鲁维酵母(K.fragilis)LFS 8611细胞生长和β D 半乳糖苷酶的合成同步.该菌株生长和产酶的最适碳源为半乳糖,乳糖次之;最适氮源为蛋白胨F403;最适培养条件为:发酵培养基的初始pH值为7.0,摇床的转速为200r/min.培养基中碳源和氮源质量浓度对菌体生物量和β D 半乳糖苷酶活力有重要影响,以12mg/mL乳糖为碳源,16mg/mL蛋白胨(F403)为氮源,在最适培养条件下培养32h后,菌体生物量和β D 半乳糖苷酶活力分别为7.56g/L和18.83U/mL.     

pH值对聚唾液酸分批发酵的影响及补料发酵

研究了不同pH值对聚唾液酸发酵过程中菌体生长和产聚唾液酸的影响.发酵初期pH自然下降时有利于菌体生长,菌体生长对数期较长,最大菌体干重可达6.9g/L;发酵中后期pH控制在6.4时有利于聚唾液酸的延续合成,合成对数期比其它pH条件下的合成对数期延长了11h.动力学特性表现为部分相关模式,而在其它pH条件下,动力学特性表现为相关模式.对聚唾液酸的流加补料发酵进行了初步研究,最终使菌体干重达到11.16g/L,聚唾液酸产量达到2.606g/L.     

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.     

嗜热芽孢杆菌2004产β-甘露聚糖酶产酶条件优化和酶学性质

对芽孢杆菌2004 β-甘露聚糖酶进行了产酶条件优化、初步纯化及其酶学性质的研究.条件优化结果:1.5 g/dL槐豆胶,2 g/dL蛋白胨,培养温度为40 ℃,250 mL三角瓶装液量为70 mL,其他因素影响不大.此最佳产酶条件下,嗜热芽孢杆菌2004 12 h的产酶水平平均为41.92 U/mL,比原来提高了30倍.培养液离心得到上清液,经硫酸铵沉淀,Sephadex G-200分子凝胶过滤和DEAE纤维素离子交换,酶比活达到722 U/mg,纯化了23.94倍,收率为35.1 %.该酶的最适反应温度为70～80 ℃;反应的最适pH值为5.8～6.0;pH值稳定范围为4.0～9.0.金属离子Mg2 和K 对酶略有激活作用.适当浓度的Mg2 不仅对酶有激活作用,还对酶的热失活具有保护作用.     

固态发酵生产细菌α-淀粉酶

采用国内α 淀粉酶生产常用菌株BF7658变异菌种 ,直接以麸皮为原料固态发酵法生产α 淀粉酶 ,得到较适宜的条件为 :培养基初始含水量 (质量分数 )为 60 % ,起始 pH自然 ,液体接种量为 5mL/kg ,3 7～ 3 9℃培养 4 8～ 60h ,三角瓶培养产酶水平可达 1 2 4 8U/ g ,浅盘培养平均产酶可达 1754U/ g .固态发酵生产细菌α 淀粉酶产酶水平为液体深层发酵法的 4～ 5倍 ,并且成本低廉 ,具有较好的经济和环境效益 .     

产胆固醇氧化酶重组大肠杆菌的发酵培养基和诱导条件的优化

在摇瓶中对产胆固醇氧化酶重组大肠杆菌的发酵培养基和诱导条件进行了优化,优化培养基为甘油10 g/L,胰蛋白胨10 g/L,酵母粉5 g/L,KH2PO4 2 g/L,K2HPO4 4 g/L,Na2HPO4·12H2O 7 g/L,(NH4)2SO4 1.2 g/L,NH4Cl 0.2 g/L,MgSO4·7H2O 1 g/L;优化的诱导条件为:对数生长中期诱导,IPTG浓度为0.3 mmol/L.在优化的培养基和优化的诱导条件下,单位菌体产酶量达745.86 U/g,菌体产酶水平达1 625.97 U/L,为优化前的700 U/L的2.3倍.     

酵母流加发酵过程中的模糊控制器

通过分析酵母发酵过程中的影响因素，建立了控制酵母发酵过程中流加速率的模糊控制器.该模糊控制器的模糊规则可以根据专家经验方便地修改以提高控制效果.实验表明在模糊控制器控制下酵母产量比恒速流加时提高了一倍.     

搅拌转速和通气量对灰树花深层发酵培养的影响

采用15L搅拌式发酵罐研究灰树花的扩大培养条件，主要考察了搅拌转速和通气量对灰树花菌丝量、胞外多糖产率、溶氧、菌丝形态以及发酵液粘度的影响．研究结果表明搅拌转速和通气量能显著影响灰树花的生长，并且通过对灰树花菌丝球的大小、形态以及胞外多糖的分泌影响发酵液的粘度．并得出较为优化的培养条件为：温度25℃，通气量0．75vvm，搅拌转速80r／min，装液量60％；在此条件下，发酵10d后，灰树花菌丝体最大得率为22．95g／L，胞外多糖的得率为1．521g／L。     

Outcome of Patients Surgically Treated for Various Forms of Hyperthyroidism with Differentiated Thyroid Cancer: Experience at an Endocrine Center in Italy

Purpose The incidence and aggressiveness of thyroid cancer associated with hyperthyroidism remains a subject of much controversy. The aim of this study was to analyze the frequency of coexisting hyperthyroidism and thyroid cancer, and to determine whether cancer becomes more aggressive with different forms of hyperthyroidism. Methods We retrospectively studied 2 449 patients assessed for hyperthyroidism between 1985 and 2001. All patients with a “cold” nodule on scintigraphy, such as those with Graves' disease and a concomitant solid nodule, underwent fine-needle aspiration biopsy (FNAB). Criteria for surgery were cytological findings indicative of malignancy, goiter with signs of tracheal or esophageal compression, side effects of antithyroid drug therapy, or Graves' disease with multiple relapses after therapy withdrawal or responsiveness to antithyroid drugs. Results Thyroid cancer was diagnosed more frequently in patients with Graves' disease (6.5%) than in those with uninodular toxic goiter (UTG) (4.4%) or multinodular toxic goiter (MTG) (3.9%). Lymph node involvement was found in 56% of the patients with Graves' disease, in 23% of those with MTG, and in none of those with UTG. Distant metastases were found in one patient with Graves' disease. Conclusions Cancers associated with Graves' disease seems to be more aggressive than those associated with MTG or UTG. Thus, we suggest that patients with Graves' disease be carefully monitored for the detection of thyroid nodules. Ultrasonography seems to be the best modality to detect such nodules.     

DTI tractography of the human brain's language pathways

Diffusion Tensor Imaging (DTI) tractography has been used to detect leftward asymmetries in the arcuate fasciculus, a pathway that links temporal and inferior frontal language cortices. In this study, we more specifically define this asymmetry with respect to both anatomy and function. Twenty right-handed male subjects were scanned with DTI, and the arcuate fasciculus was reconstructed using deterministic tractography. The arcuate was divided into 2 segments with different hypothesized functions, one terminating in the posterior superior temporal gyrus (STG) and another terminating in the middle temporal gyrus (MTG). Tractography results were compared with peak activation coordinates from prior functional neuroimaging studies of phonology, lexical-semantic processing, and prosodic processing to assign putative functions to these pathways. STG terminations were strongly left lateralized and overlapped with phonological activations in the left but not the right hemisphere, suggesting that only the left hemisphere phonological cortex is directly connected with the frontal lobe via the arcuate fasciculus. MTG terminations were also strongly left lateralized, overlapping with left lateralized lexical-semantic activations. Smaller right hemisphere MTG terminations overlapped with right lateralized prosodic activations. We combine our findings with a recent model of brain language processing to explain 6 aphasia syndromes.     

在模型化基础上对青霉素补料分批发酵过程中经济效益的预测

在研究发酵动力学、生物过程反应计量学、代谢流分布和代谢流控制的基础上 ,结合专家知识、实验及生产数据 ,对生产规模的青霉素反复补料分批发酵 (即半连续发酵 )的全过程进行了数学模拟 ,建立了一套包含 2 5个过程变量和一系列经济学变量的改良机理模型 .根据这一模型 ,预测了一些工艺参数和经济学参数的改变对发酵过程经济效益的影响 ,发现在保持最大生物质浓度、稀释速率 ,及其它工艺条件不变的情况下 ,中间放料间隔时间、初始发酵液体积以及葡萄糖和电力的价格对经济效益产生较大的影响 .模拟运算结果表明 ,葡萄糖和电力的价格降低 5 % ,全年利润可分别提高 6 .35 %和 3.75 % ;中间放料间隔时间由 2 4h缩短为 12h和 1h(接近于连续发酵 ) ,全年利润可分别提高 7.2 2 %和 14 .11% ;初始发酵液装罐体积分数由 75 %提高到 85 % ,全年发酵利润可提高 5 .4 8% .     

脊柱骨巨细胞瘤与脊索瘤局部侵袭范围的组织学研究

目的 观察脊柱骨巨细胞瘤和脊索瘤在不同类型瘤周组织中的侵袭距离,为两者手术切除范围的确定提供组织学依据.方法 将7例脊柱骨巨细胞瘤与10例脊柱脊索瘤患者的手术切除标本,按瘤周组织类型的不同,分为骨皮质组、软骨组、骨松质组、肌肉组、脂肪组和瘢痕组,分别测量各组标本肉眼边界(MSM)、组织学边界(HLM)与分子边界(MCM),并进行各边界组内比较和MSM.HLM、MSM-MCM组间比较.结果 除骨皮质组与软骨组外,其余4组边界组内比较存在统计学差异(P＜0.05).骨松质组、脂肪组、肌肉组和瘢痕组MSM-HLM,MSM-MCM差值显著大于软骨组和(或)骨皮质组同一种差值(P＜0.05),边界差值的最大值分别为11.68 mm,8.64 mm,13.08 mm和9.98 mm.结论 脊椎可被视为一个解剖间室,对局限于该间室的骨巨细胞瘤和脊索瘤,应首选全椎切除;在瘤周松质骨、椎旁肌肉和脂肪组织内行广泛性切除时,切除范围应达到肿瘤肉眼所见范围外1.3 cm,1.5 cm和1.0 cm;重视首次手术切除彻底性,复发再手术时,瘤旁瘢痕切除范围至少应达到肉眼所见范围外1.0 cm.     

Speaking of which: dissecting the neurocognitive network of language production in picture naming

The noninvasive methods of cognitive neuroscience offer new possibilities to study language. We used neuronavigated multisite transcranial magnetic stimulation (TMS) to determine the functional relevance of 1) the posterior part of left superior temporal gyrus (Wernicke's area), 2) a midportion of Broca's area (slightly posterior/superior to apex of vertical ascending ramus), and 3) the midsection of the left middle temporal gyrus (MTG), during overt picture naming. Our chronometric TMS design enabled us to chart the time points at which neural activity in each of these regions functionally contributes to overt speech production. Our findings demonstrate that the midsection of left MTG becomes functionally relevant at 225 ms after picture onset, followed by Broca's area at 300 ms and Wernicke's area at 400 ms. Interestingly, during this late time window, the left MTG shows a second peak of functional relevance. Each area thus contributed during the speech production process at different stages, suggesting distinct underlying functional roles within this complex multicomponential skill. These findings are discussed and framed in the context of psycholinguistic models of speech production according to which successful speaking relies on intact, spatiotemporally specific feed forward and recurrent feedback loops within a left-hemispheric fronto-temporal brain connectivity network.     

核黄素产生菌的补料发酵

通过分析核黄素产生菌E .ashbyii在发酵过程中菌体量、糖质量浓度、pH值、核黄素质量浓度和粘度的变化及菌体形态与核黄素生物合成之间的联系 ,发现 pH值对菌体生长有显著的影响 ,菌体浓度过高会导致发酵液粘度过大 ,通风供氧困难 ,菌体活力下降 .采用先低浓度培养菌体 ,然后根据发酵液 pH值的变化进行多次适量补料 ,不仅可以控制菌体的过度增殖 ,缓解通风供氧不足造成的困难 ,而且还可以保持菌体活力 ,增加核黄素的产量 .     

红酵母类胡萝卜素发酵助剂的筛选及应用

研究了几种促进剂和前体对红酵母RY－98类胡萝卜素发酵的影响，从中选出了番茄汁、花生油和核黄素3种对红酵母生长及类胡萝卜素合成具有显著促进作用的发酵助剂，并确定了适宜的用量。应用试验和发酵过程动态分析表明，这3种发酵助剂增产效果明显。当同时添加番茄汁3mL/L、花生油1.2mL/L和核黄素3.5mg/L时，菌体量、类胡萝卜素质量分数和产量可分别比对照组提高39.4%、32.8%和85.1%，且对发酵过程菌体生长及生理代谢规律无不良影响。