葡聚糖酶高产菌株的诱变筛选及其摇瓶发酵条件的初步研究

从土壤中分离出的一株产葡聚糖酶酶活31 U／ml的野生菌株,经UV、^60Co、LiCl诱变筛选后得到了产葡聚糖酶高产菌株SB126,经发酵条件优化试验后检测其酶活达到85U／ml,较野生菌株提高了近两倍。葡聚糖酶摇瓶较适发酵条件为：装量30ml(250m1)、转速180r／min、pH7．0、温度30℃、接种量8％、发酵周期5d。     

产木聚糖酶菌株的选育及其液体发酵条件

从土壤中筛出一株生长快产木聚糖酶活力较高的黑曲霉(Aspergillus niger)C—2菌株,经紫外线和甲基磺酸乙酯诱变,获得一突变株An—76,其生长减缓,孢子形成能力减弱,产生的术聚糖酶和β—木糖苷酶活力分别可达353．61U／ml和4．51U／ml。测定了An—76的正常产酶曲线,研究了麸皮、氮源、碳源及半纤维素浓度对产酶的影响。最适培养条件为：起始pH6.0、28℃、96h。酶的最适作用条件为50—55℃,pH4．8,在pHl．2—11．4范围内稳定。酶的热稳定性较差,55℃保温1小时,剩余活力为40％。只有在含木糖苷类物质存在时,An—76才大量合成木聚糖酶。     

响应面法优化Burkholderiasp．SYBCLIP—Y发酵产低温脂肪酶条件的研究

用响应面法对Burkholderiasp．SYBCLIP—Y液体发酵产低温脂肪酶的发酵条件进行了快速优化。首先利用Plackett—Burman设计对影响其产酶相关因素进行评估并筛选出具有显著效应的三个因素：牛肉膏,橄榄油,TritonX-100;用最陡爬坡路径逼近最大产酶区域后,利用响应面中心组合设计对显著因素进行优化,确定出牛肉膏,橄榄油,TritonX-100的最佳浓度分别为：牛肉膏31．8g／L、橄榄油21mL／L、TritonX-10036．55mL／L,优化后脂肪酶的酶活达到61．52U／mL,是优化前的2．62倍。     

枯草杆菌SOD高产菌株的诱变选育及产酶条件研究

本实验采用低能氮离子注入技术对枯草芽孢杆菌(Bacillus subtilis)进行辐照诱变处理,选育出一株SOD高产菌株(编号为BsB8)。其生物量略高于出发菌株、SOD产量达3439．3U／g湿菌体,为实验出发菌的1．88倍。该菌株最佳产酶条件为：起始pH为8．2,装液量为150ml／500ml,接种量为1．5％;添加Mn^2 的培养基可显著提高SOD酶活。     

霉菌α—糖苷酶固态发酵的研究

研究了一株产α-糖苷酶能力较高的黑曲霉菌株H－8的固态发酵的较佳产酶条件。结果表明：当麸皮加水比为1：1．00、混料装量10g（500ml三角瓶）、接种量2环、35℃培养48h后,酶活力对干麸曲计可达496.89U／g。     

微生物法生产普鲁兰酶的研究

对产普鲁兰酶的出发菌株进行筛选和诱变,使酶活从22．10u／ml提高到了40．77u／ml,酶活提高了84．4％。随后对菌体产酶培养基进行了确定和优化,得到最佳发酵培养基,在此培养基下,菌体产酶达46．76u／ml,为原酶活的114．7％。     

真空微波诱变结合化学诱变选育酸性纤维素酶高产菌株

以酸性纤维素酶产生菌绿色木霉（Trichoderma viride）WL0512作为原始出发菌株,首先经自然分离筛选出一株产酶较稳定的菌株TVN-18,其羧甲基纤维素酶活（CMC酶活）达2765．8U／g,滤纸酶活（FPA酶活）达48．5U／g。再经真空微波和甲基磺酸乙酯（EMS）逐级诱变处理,获得了一株高产、稳产酸性纤维素酶的E6—1菌株,其CMC酶活达4396．6U／g,FPA酶活达126．0U／g,分别是菌株TVN-18的1．59倍和2．60倍。通过对固态发酵培养基麸皮和稻草比例、料水比以及初始pH值的优化,突变株的产酶能力进一步得到提高,其产的CIVIC酶活和FPA酶活分别提高了22．3％和22．4％。     

黑曲霉产纤维素酶的研究

从土壤中筛选获得一株产纤维素酶的优良菌株黑曲霉Asp．n－21,采用固体培养产生纤维素酶,产酶活力FPA137U／g干曲、GMCase320～388U／g干曲、β－葡萄糖苷酶84～149U／g干曲,对培养基成份进行优化,并分析其酶系组成,该菌所产酶可作为饲料用酶。     

植物乳杆菌RS2-2发酵法生产乳酸脱氢酶

筛选到一株产乳酸脱氢酶的植物乳杆菌RS2-2（Lactobacillus plantarum RS2-2）。产酶条件研究表明,以蔗糖为碳源,酵母膏为氮源,起始pH6．5左右,30℃发酵,接种量为10％,产酶量最高。在25L发酵罐放大实验,并通过流加NaOH控制发酵过程中的pH,所产乳酸脱氢酶比活力为18．73U／mg,菌重量为lg／L以上。粗酶液经分离纯化得到成品酶液,比酶活提高了51．7倍,达到1210、7U／mg,总收率为30、2％。用凝胶过滤检测其分子量分布,分子量约为85,000Da,酶纯度为88％。     

两株保加利亚乳杆菌产β-半乳糖苷酶条件的优化及酶活力测定

目的优化2株保加利亚乳杆菌产β-半乳糖苷酶的培养条件,测定其在最适于产酶条件下的酶活力,并初步观察酶活力稳定性。方法测定2株保加利亚乳杆菌——来源于酸奶的wch9901和标准菌株1．1480在不同培养时间、培养基碳源、摇床转速、气体环境下产生的β-半乳糖苷酶的活力,优化产酶条件。测定2株菌在其最适产酶条件下的酶活力。将制得的1．1480粗酶液分别置冰浴和20℃水浴,每隔1h测定一次粗酶液中β-半乳糖苷酶活力,观察酶活力稳定性。结果2株保加利亚乳杆菌的最适产酶条件分别为：1．1480于30℃有氧静止培养36h;wch9901于37℃厌氧静止培养18h,培养基含乳糖。在最适产酶条件下,1．1480的酶活力为0．321NLU／ml粗酶液,比酶活为2．469NLU／mg蛋白;wch9901的酶活力为0．401NLU／ml粗酶液,比酶活为6．169NLU／mg蛋白。1．1480粗酶液于冰浴可稳定保存6h,于20℃水浴可稳定保存5h。结论wch9901与1．1480的最适产酶条件有所差异,前者产酶迅速,产酶量多。     

Production and potential applications of a xylanase from a new strain of Streptomyces cuspidosporus

Enzyme production by a new mesophilic Streptomyces isolate was investigated which grew optimally on 1% (w/v) xylan and 10% (w/v) wheat bran at pH 7 and 37 °C. Xylan induced only CMCase (0.29 U/ml) besides xylanase (22–35 U/ml, 40–49 U/mg protein). Wheat bran induced xylanase (105 U/ml, 17.5 U/mg protein), CMCase (0.74 U/ml), -xylosidase (0.009 U/ml), -glucosidase (0.026 U/ml), -L-arabinofuranosidase (0.049 U/ml), amylase (1.6 U/ml) and phytase (0.432 U/ml). The isolate was amenable to solid state cultivation and produced increased levels of xylanase (146 U/ml, 28 U/mg protein). The pH and temperature optima of the crude xylanase activity were 5.5 and 65 °C respectively. The pI was 6.0 as determined by PEG precipitation. The crude enzyme was applied in treatment of paper pulp and predigestion of poultry feed and was found to be effective in releasing sugars from both and soluble phosphorus from the latter.     

Improved production of an enzyme that hydrolyses raw yam starch by Penicillium sp. S-22 using fed-batch fermentation

A newly isolated strain, Penicillium sp. S-22, was used to produce an enzyme that hydrolyses raw yam starch [raw yam starch digesting enzyme (RYSDE)]. The enzyme activity and overall enzyme productivity were respectively 16 U/ml and 0.19 U/ml h in the batch culture. The enzyme activity increased to 85 U/ml by feeding of partially hydrolyzed raw yam starch. When a mixture containing partially hydrolyzed raw yam starch and peptone was fed by a pH-stat strategy, the enzyme activity reached 366 U/ml, 23-fold of that obtained in the batch culture, and the overall productivity reached 3.4 U/ml h, which was 18-fold of that in the batch culture.     

黑曲霉生产菊粉酶工艺条件的研究

从徐州市沛县河口镇秦庄村牛蒡种植基地取得的土壤样本中,筛选出产菊粉酶的菌株,对从土壤中分离到的40株产菊粉酶的各类微生物进行酶活的测定。通过透明圈法初筛及摇瓶复筛,获得产菊粉酶能力较高的霉菌菌株3株,为C122803、D081506和D081513,这3株菌株的酶活分别达到：C122803：1.411 U/ml;D081506 ：1.895U/ml;D081513 ：1.792U/ml。其中D081506的酶活最高,为1.895U/ml;对D081506号黑曲霉产菊粉酶的发酵条件进行了研究,确定了优化的发酵条件为：牛蒡汁2%,酵母膏1.6%,(NH4)2SO41.6%,NaCl 0.5%,K2HPO4 0.5%,pH 5.0,在27℃、140 r/min条件下,摇瓶培养24h, D081506酶活力为2.958U/ml,酶活力提高56.09%。     

Preparation and characterisation of immobilised metal ion hollow-fibre polysulphone membranes. Their application in high-speed pectic enzyme fractionation

Chelating hollow-fibre membranes were prepared from epoxy-activated polysulphone microfiltration fibres by introducing iminodiacetic acid (IDA) groups in the presence of dimethyl sulphoxide. Fibres with 160, 350 and 620 μmol epoxy groups/ml provided ligand densities of 69, 134 and 203 μmol IDA/ml and pure water fluxes of 7.8, 5.8 and 0.42 cm/min, respectively. However, lysozyme capacity was close to 4 μmol/ml for all fibres. Adsorption isotherms for lysozyme and pectinesterase did not fit Langmuir-type curves and the existence of two types of ligand (A and B) with different accessibility to proteins was assumed. For pectinesterase, maximum capacities of 5100 and 2900 U/ml and dissociation constants of 25 and 316 U/ml were found, respectively, for ligands A and B. For lysozyme, maximum capacities were 2.9 and 0.9 μmol/ml and dissociation constants 5.0 and 102 μM, respectively, for said ligands. A cartridge assembled with IDA hollow fibres had a dynamic capacity for pectinesterase of 7509 U/ml. Productivity of this cartridge for pectic enzyme fractionation was 750 pectinesterase U/ml min, far higher than that obtained with a chelating soft gel (81 pectinesterase U/ml min).     

Effects of high-density lipoproteins on thrombin-induced platelet aggregation

Using new-developed method of aggregates radius measurement in suppression of washed human platelets, it has been shown, that HDL, in concentration of 190 ug/ml and in higher ones, totally inhibited aggregation, induced by 0.075 U/ml thrombin. For the same effect on aggregation, induced by 0.225 U/ml thrombin, HDL in concentration of 1320 ug/ml were needed.     

Optimization of cellulase production in batch fermentation byTrichoderma reesei

Maximum cellulase production was sought by comparing the activities of the cellulases produced by differentTrichoderma reesei strains andAspergillus niger. Trichoderma reesei Rut-C30 showed higher cellulase activity than otherTrichoderma reesei strains andAspergillus niger that was isolated from soil. By optimizing the cultivation condition during shake flask culture, higher cellulase production could be achieved. The FP (filter paper) activity of 3.7 U/ml and CMCase (Carboxymethylcellulase) activity of 60 U/ml were obtained from shake flask culture. When it was grown in 2.5L fermentor, where pH and DO levels are controlled, the Enzyme activities were 133.35 U/ml (CMCase) and 11.67 U./ml (FP), respectively. Ammonium sulfate precipitation method was used to recover enzymes from fermentation broth. The dried cellulase powder showed 3074.9 U/g of CMCase activity and 166.7 U/g of FP activity with 83.5% CMCase recovery.     

Scale-up of an alkaline protease from Bacillus pumilus MTCC 7514 utilizing fish meal as a sole source of nutrients

Fish meal grades SL1 and SL2 from Sardine (Sardinella longiceps) and NJ from Pink Perch (Nemipterus japonicas) were evaluated as a sole source of carbon and nitrogen in the medium for alkaline protease production by Bacillus pumilus MTCC 7514. The analysis of the fish meal suggests that the carbon and nitrogen contents in fish meal are sufficient to justify its choice as replacement for other nutrients. Protease production increased significantly (4,914 U/ml) in medium containing only fish meal, compared with the basal medium (2,646 U/ml). However, the elimination of inorganic salts from media reduced the protease productivity. In addition, all the three grades of fish meal yielded almost the same amounts of protease when employed as the sole source of carbon and nitrogen. Nevertheless, the best results were observed in fish meal SL1 medium. Furthermore, protease production was enhanced to 6,966 U/ml and 7,047 U/ml on scaling up from flask (4,914 U/ml) to 3.7 and 20 L fermenters, respectively, using fish meal (10 g/l). Similarly, the corresponding improvement in productivities over flask (102.38 U/ml/h) was 193.5 and 195.75 U/ml/h in 3.7 and 20 L fermenters, respectively. The crude protease was found to have dehairing ability in leather processing, which is bound to have great environmental benefits.     

Enhanced production of a thermostable xylanase from Streptomyces sp. QG-11-3 and its application in biobleaching of eucalyptus kraft pulp

A thermostable and cellulase-free xylanase has been produced from Streptomyces sp. QG-11-3 in solid substrate fermentation using wheat bran and eucalyptus kraft pulp as the prime solid substrates. The maximum xylanase yield obtained using these two substrates were 2360 U/g and 1200 U/g dry solid substrate at substrate:moisture ratios of 1:3 and 1:2.5, respectively. In immobilized cell system using polyurethane foam (PUF) and three nonwoven fabrics, namely, polyester, silk, and cotton, the xylanase yields were enhanced by 2.5-fold (203 U/ml), 1.91-fold (155 U/ml), 1.54-fold (125 U/ml), and 1.47-fold (119 U/ml), respectively, compared to the xylanase yield in liquid-batch fermentation (81 U/ml). In the biobleaching experiments, the xylanase dose of 3.5 U/g moisture free pulp exhibited the optimum bleach boosting of eucalyptus kraft pulp at pH 8.5 and 50 degrees C after 2 h of treatment. When xylanase treated pulp was subsequently treated with 4.5% chlorine, it resulted in reduction of kappa number by 25%, enhanced the brightness (%ISO) by 20% and improved the pulp properties such as tensile strength and burst factor by up to 63% and 8%, respectively.     

Modulation by novobiocin of sister-chromatid exchanges induced by tumor necrosis factor in human lymphocytes.

The effect of the antibiotic novobiocin on human recombinant tumor necrosis factor (TNF)-induced sister-chromatid exchanges (SCEs) were examined in human peripheral blood lymphocytes. TNF, when introduced in a dose range of 10-1000 U/ml at the initiation of culture, was found to cause a significant increase in SCE frequency. The simultaneous addition of TNF and novobiocin (25 micrograms/ml) in the assay resulted in no increase of SCE frequency. Delayed (for 24 h) addition of novobiocin suppressed the induction of SCEs by 50, 100 and 500 U/ml but not by 1000 U/ml of TNF.