采用D-饱和试验设计确定产植酸酶菌株代谢的营养条件

本研究对黑龙江海伦市种植大豆作物的黑色土壤中筛选得到的产植酸酶菌株(穗霉属26-13-4)的最佳营养条件进行优化。针对发酵培养基中的碳源、氮源、麸皮的添加量进行D-饱和最优化设计, 拟合三者之间的方程, 并确定最优化添加量。     

产植酸酶菌株的筛选及产酶条件的研究

通过初筛和复筛,得到一株产植酸酶较高的黑曲霉AN00101菌株,并对该菌种的产酶条件进行了研究.结果表明:配制加水量为35%的麸皮固体培养基,在37℃培养114h,用3%CaCl2进行提取,每g固体发酵物酶活高达1.3×104IU.经L9(34)正交实验表明,硫酸铵和硫酸镁对产酶有显著的促进作用,适宜添加量分别为4%和0.3%.     

产植酸酶菌株的分离筛选研究

研究了产植酸酶菌株分离筛选的方法 ,筛选得到分别适宜液态发酵和固态发酵的酶活力较高的黑曲霉XH和黑曲霉PA两菌株。在液态发酵KH2 PO4 对植酸酶产生有促进作用 ,其最佳用量为 8mg %;在固态发酵KH2 PO4 的添加对植酸酶的产生无明显促进作用。 2菌株所产植酸酶具有相似的酶学性质 ,最适作用温度为 5 5℃ ,当底物 pH为 3～ 5具有较高的酶活力。     

中性植酸酶高产菌株的筛选及产酶条件研究

以地衣芽孢杆菌为原始出发菌株,用紫外线反复诱变,最终获得一株中性植酸酶高产菌株B.licheniformis LL8,其酶活性约为原始出发菌株的2倍。该菌株具有稳定的遗传性能。通过单因素试验和正交试验对发酵条件进行了优化,当培养温度为55℃,pH为7.5,接种量为10%时,经30h培养后,中性植酸酶活力最高达到2268.4U/mL。     

激光诱变筛选高产植酸酶黑曲霉菌株的研究

用1.06um,15KHz高重复率声光调QNd：YAG激光以20W和15W的辐照功率分别照射产植酸酶的黑曲霉孢子液和原生质体液不同时间,检定再生菌落数并经透明圈初筛和摇瓶复筛,测定诱变菌株所产植酸酶酶活。结果表明：20W的功率辐照黑曲霉孢子1′以上,致死率近乎100…,15W功率辐照原生质体液20″-4′,存活率在8.571％-117.14％之间,正变率在27.45％-100％之间,正变辐度60.69％-124.96％,说明原生质体比孢子耐受激光诱变的能力强,并且诱变效果好。筛选到了一株酶活提高达3.75倍的单个变异菌株。     

植酸酶对斑点叉尾鲴生长性能及磷当量的研究

试验旨在研究饲料中添加植酸酶对斑点叉尾 [初始平均体重(1.70±0.04) g]生长性能及植酸磷利用的影响, 确定植酸酶的磷当量。试验采用单因素试验设计, 以 Ca(H2PO4)2提供外源无机磷, 同时添加不同浓度植酸酶, 试验设计为 10 个处理组, 分别为 1 个对照试验组、4 个无机磷试验组(0.3%、0.5%、0.8%、1.2%)和 5 个植酸酶试验组(300、500、1000、1500、2000 U/kg), 每个处理 3 个重复, 每个重复 30 尾鱼。通过折线模型确定植酸酶的最佳添加量; 通过回归分析, 建立响应指标(特定生长率、椎骨磷)与外源磷添加量之间的线性关系, 进而确定植酸酶的磷当量值。结果表明: (1)添加植酸酶的处理组与对照组相比, 增重率、特定生长率、蛋白质效率、肥满度均有显著提高(P<0.05), 饲料系数、肝体比、脏体比均有下降(P<0.05), 成活率各处理组没有显著差别(P>0.05); 鱼体粗蛋白、粗灰分、钙、磷及椎骨粗灰分、钙、磷均有显著提高(P<0.05), 鱼体粗脂肪含量有所下降。(2)无机磷添加水平与响应指标之间线性关系如下: Y1=0.2714X+2.294(X-无机磷, Y1-特定生长率, R2=0.9238), 300、500、1000、1500 和 2000 U/kg 植酸酶分别可替代 1 kg 饲料中 0.13%、0.57%、0.76%、1.46% 和 1.35% 的 磷 酸 二 氢 钙 , 等 效 于 添 加 了 0.03% 、 0.14% 、 0.19% 、 0.36% 和 0.33% 的 有 效 磷 ;Y2=0.8737X+5.1028(X-无机磷, Y2-椎骨磷, R2=0.9638), 300、500、1000、1500 和 2000 U/kg 植酸酶分别可替代1 kg 饲料中 0.47%、1.11%、1.18%、1.38%和 1.41%的磷酸二氢钙, 等效于添加了 0.12%、0.27%、0.29%、0.34%和 0.35%的有效磷。在试验条件下, 添加 1000―2000 U/kg 植酸酶能有效改善斑点叉尾 生长性能, 有利于营养物质在鱼体中的沉积, 促进骨骼矿化。以特定生长率为响应指标, 植酸酶最佳添加量为 1435 U/kg等效于添加了 0.37%的有效磷; 以椎骨磷为响应指标, 植酸酶最佳添加量为 1226 U/kg 等效于添加了 0.33%的有效磷。     

青霉LQ-7植酸酶产生条件研究

筛选到一株植酸酶高产菌株 ,对其进行诱变并研究了该菌株产植酸酶的最适条件。优化产酶液体培养基组成为 :3%可溶性淀粉 ,0 5 %蛋白胨 ,0 5 %NH4NO3 ,0 0 5 %MgSO4·7H2 O ,0 0 5 %FeSO4·7H2 O ,0 0 0 1%MnSO4·4H2 O ;发酵培养基的起始pH为 6 5 ,接种量 4%的条件下 30℃ ,135r min培养 72h可获得较高的植酸酶产量。少量 (2 5mg)植酸盐的加入对植酸酶有促进作用 ,但过量 (10mg以上 )时则会产生抑制作用。     

包被工艺条件对植酸酶热稳定性的影响

考察了不同种类的糖、盐及浓度在干热及湿热的情况下对残存酶活的影响,通过正交实验,确定包被工艺优化条件为：以多孔淀粉为载体,采用流化干燥法取得;包被中蔗糖的添加量为40mg／g酶,氯化钠包覆用量为淀粉质量的10％,明胶作外包被,用量为淀粉质量的1.5％,得到包埋颗粒,其包埋率为82．8％,包埋后,水分活度大于0．35时,包被酶在干热的情况下残存酶活比原酶有较大的提高,残存酶活提高8．7％,湿热的情况下,残存酶活提高58．3％,胃蛋白酶对其的损坏作用也明显减小.     

植酸酶基因工程研究进展

植酸酶是催化植酸及植酸盐水解成肌醇和无机磷酸的一类酶的总称.添加于食品和饲料中,能消除植酸所引起的抗营养作用,可提高蛋白质和矿物质的生物利用率.对植酸酶的生物学特性、基因结构和基因工程的研究进展做了综述.     

雌二醇对蒙古绵羊输卵管上皮细胞内β-防御素(sBD-1)表达的影响

为了探索雌二醇与β-防御素(sBD-1)表达量的关系,体外模拟蒙古绵羊(Ovis aries)生理周期.用添加雌二醇浓度10-11、10-10、10-9、10-8、10-7、10-6 mol/L的培养液及不添加雌二醇的培养液(即对照组)分别培养蒙古绵羊输卵管上皮细胞,作用24 h、48 h、72 h后,提取细胞总RNA,应用SYBR Green I荧光定量RT-PCR法进行定量分析.结果显示.添加不同浓度雌二醇培养的输卵管上皮细胞中sBD-1的相对表达量0.000 740～0.001 758,与对照组0.000 190之间差异显著(P＜0.05),且小于10-8 mol/L雌二醇添加浓度与sBD-1基因相对表达量变化基本呈正相关.此结果为进一步研究雌激素参与机体防御功能奠定了基础.     

Efflux of Red Cell Water into Buffered Hypertonic Solutions

Buffered NaCl solutions hypertonic to rabbit serum were prepared and freezing point depressions of each determined after dilution with measured amounts of water. Freezing point depression of these dilutions was a linear function of the amount of water added. One ml. of rabbit red cells was added to each 4 ml. of the hypertonic solutions and after incubation at 38°C. for 30 minutes the mixture was centrifuged and a freezing point depression determined on the supernatant fluid. The amount of water added to the hypertonic solutions by the red cells was calcuated from this freezing point depression. For each decrease in the freezing point of -0.093°C. of the surrounding solution red cells gave up approximately 5 ml. of water per 100 ml. of red cells in the range of -0.560 to -0.930°C. Beyond -0.930°C. the amount of water given up by 100 ml. of red cells fits best a parabolic equation. The maximum of this equation occurred at a freezing point of the hypertonic solution of -2.001°C. at which time the maximum amount of water leaving the red cells would be 39.9 ml. per 100 ml. of red cells. The data suggest that only about 43 per cent of the red cell water is available for exchange into solutions of increasing tonicity.     

地木耳对蕃茄生长的影响

地木耳是蓝藻门植物,具有生物固氮作用,分别以地表施加、土中施加、浸泡材料与酶提取液的方式在番茄的营养生长期和花蕾期施加,均有增肥效应。地表施加效果最好,且营养生长期施加的作用大于花蕾期;地木耳的用量随每一植株生活的环境不同而异。只要使地木耳保持活力,白天进行光合作用,夜间便可进行固氮作用,它可以持续地进行生物固氮,且具有累加效应。     

Ionic interfaces and diphtheria toxoid interactions

We present here a systematic study on the purification of the diphtheria toxoid (Dtxd) produced at the Instituto Butantan, by adding only one step on the entire process of its production. Aliquots of 1.0 ml of Dtxd were added to an equal amount of Q-Sepharose previously equilibrated with 500 mM Tris, pH 5.0-9.0 (increments of 0.5 pH units). The best condition for the Dtxd monomer adsorption was achieved at pH 9.0. The best condition for desorption was obtained with 300 mM NaCl. After studying the gel binding capacity for Dtxd, a column (C20/20) equilibrated with 500 mM Tris, pH 9.0, was prepared. The purification factor for Dtxd was 1.5. The final recovery of Dtxd was 68.75%, with 90.31% purity. The process methodology presented here is a very realistic sequence of separation steps, which is perfectly compatible with the production requirements. Vaccination with "toxoid highly purified toxin" is known to confer a strong immunity on people in the absence of undesirable reactions, which led experts of European Pharmacopoeia to recommend its use both for children and adult vaccination.     

Optimized enzymatic synthesis of geranyl butyrate with lipase AY from Candida rugosa

Response surface methodology (RSM) and five-level, five-variable central composite rotatable design (CCRD) were used to evaluate the effects of synthetic variables, such as reaction time (1-9 h), temperature (25-65 degrees C), enzyme amount (10-50%), substrate molar ratio of geraniol to tributyrin (1:0.33-1:1), and added water amount (0-20%) on molar percent yield of geranyl butyrate, using lipase AY from Candida rugosa. Reaction time and temperature were the most important variables and substrate molar ratio had no effect on percent molar conversion. Based on contour plots, optimum conditions were: reaction time 9 h, temperature 35 degrees C, enzyme amount 50%, substrate molar ratio 1:0.33, and added water 10%. The predicted value was 100% and actual experimental value was 96.8% molar conversion. (c) 1996 John Wiley & Sons, Inc.     

Synthesis of ovalbumin and globin during simultaneous translation of their mRNA in the presence of tRNA of different origin in the cell-free protein-synthesizing system from wheat germ

Competition among ovalbumin and globin mRNA under their simultaneous translation in the tRNA-dependent cell-free system from wheat germ was studied. One of the mRNAs was added to samples in a constant amount, that provided 50% protein synthesis level of the maximum, other--in increasing amount (from 0 to the maximum). The ration of ovalbumin and globin synthesis rates has been shown to depend essentially (1.5-2 fold) on the nature of tRNA, being added to the system. The obtained data suggest that functional adaptation of tRNA is a part of the mechanism of protein biosynthesis regulation and it is possible to modulate some mRNAs translation selectivity on the elongation by different sets of tRNA.     

New insulating particleboards prepared from mixture of solid wastes from tissue paper manufacturing and corn peel

New composite boards with low-thermal conductivity produced from a mixture of solid wastes from tissue paper manufacturing (solid waste TPM) and corn peel have been developed. The effects of solid waste TPM/corn peel ratio on the properties of the boards were investigated and the possibility of using recycled polystyrene packaging foam as a laminating agent to improve the quality of the boards was also evaluated. Our results show that the density of the particleboards decrease with increasing the amount of corn peel added in the mixture, leading to a decrease in thermal conductivity of the final product. In contrary, larger amount of solid waste TPM added in the mixture produced stronger boards. The lamination of recycled polystyrene on the surface of particleboards improves the mechanical properties and reduces the thickness swelling of the boards. The best improvement in mechanical properties and swelling resistance could be achieved when 15% polystyrene (w/v) was coated on the surface of the boards.     

Simultaneous determination of NO and NO in fisheries wastewaters high in chloride concentration by capillary ion electrophoresis with direct UV detection

The simultaneous determination by capillary ion electrophoresis of nitrate and nitrite in wastewaters with chloride concentrations of 15 to 23g/l is described. Chloride concentrations over 200mg/l hampered the determination; thus, lead, mercuric and silver acetate were used to precipitate chloride. Silver acetate added in 1.5 times the stoichiometric amount for AgCl formation gave the best results in terms of nitrate and nitrite peak resolution.     

The amounts of adenosine di- and triphosphates bound to H-meromyosin and the adenosinetriphosphatase activity of the H-meromyosin-F-actin-relaxing protein system in the presence and absence of calcium ions. The physiological functions of the two routes of myosin adenosinetriphosphatase in muscle contraction.

The rates of the ATPase [EC 3.6.1.3] reaction of the H-meromyosin-F-actin-relaxing protein system were measured in 2 mM MgCl2, 50mM KC1, and 10mM Tris-HC1 at pH 7.8 and 20 degrees in the presence and absence of 0.05-0.1 mM Ca2+ ions. The concentrations of H-meromyosin (HMM) and the F-actin-relaxing protein (F-A-PR) complex were 3.4 and 3 mg/ml, respectively, and the ATPase reaction was coupled with 4 mg/ml of pyruvate kinase [EC 2.7.1.40] and 1 or 20 mM phosphoenolpyruvate to regenerate ATP. The amount of ADP bound to HMM during the ATPase reaction was determined by measuring the amount of ADP remaining in the reaction mixture. The amount of ATP bound to HMM was determined by subtracting the amount of bound ADP from the total amount of nucleotides bound to HMM, which was measured by a rapid flow-dialysis method. The following results were obtained. 1. The ATPase activity of the HMM-F-A-RP system increased linearly with increase in the amount of ATP added, and was independent of the presence of 0.05 mM Ca2+, when the amount of ATP added was less than 1 mole/mole of HMM. In the presence of 0.05 mM Ca2+, the ATPase activity reached a maximal level when 1.2-1.5 mole of ATP was added per mole of HMM, and maintained this level even at 3 moles of added ATP/mole of HMM. In the presence of 3mM EGTA, the ATPase activity decreased with increase in the amount of ATP added, from 1.5 to 3 moles of ATP/mole of HMM, and reached the level of the HMM ATPase reaction at 3 moles of added ATP/mole of HMM. Similar results were observed when the concentration of HMM was maintained at 3.4 mg/ml and the concentration of the F-A-RP complex was decreased from 3 to 1 or 0.5 mg/ml.     

The effect of incubation conditions on the laboratory measurement of the methane producing capacity of livestock wastes

The effects of incubations conditions (dilution, mixing, incubation time and inoculum amount and origin) on the determination of the maximum methane producing capacity (B(0)) from various livestock slurries were evaluated. For this purpose, the methane yields of different livestock slurries were determined using batch anaerobic incubations performed at 30 degrees C as regard these different conditions. The B(0) and the methane (CH(4)) generation as a function of time were used to study the processes and to determine the best incubation conditions. Methanogenesis was identified as the major rate-limiting step during the anaerobic degradation of slurries, probably due to inhibition by volatile fatty acids. In some cases, high free NH(3) concentrations were suspected to inhibit the hydrolysis process. The addition of inoculum and/or the dilution of the substrate reduced the inhibition and allowed to reach the B(0) more rapidly. However, the addition of inoculum must be minimized to reduce the possible errors made by considering a similar production by the inoculum with and without the substrate. All experiments performed during this study allowed to define the incubation conditions required for the determination of the B(0) from livestock slurries. Applying these conditions, the B(0) values determined for swine slurries varied from 244 to 343L CH(4)kg V(added)(-1), from 204 to 296L CH(4)kg V(added)(-1) for dairy cattle slurries and equalled 386 and 319L CH(4)kg VS(added)(-1) respectively for calves and duck slurries.