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.     

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。     

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%.     

氮添加对樟子松人工林土壤细菌磷酸酶编码基因丰度的影响

为研究氮添加影响森林土壤有机磷矿化的微生物调控机制,分析了10年的野外氮添加(100 kg N ha^-2year^-1)对沙地樟子松人工林土壤微生物中编码酸性磷酸单酯酶、碱性磷酸单酯酶和植酸酶的功能基因(phoC、phoD和appA)丰度及相关酶活性和土壤理化性质的影响。结果表明,氮添加使樟子松人工林土壤中酸性和碱性磷酸单酯酶活性分别下降了18.09%和55.29%,植酸酶活性下降了41.88%。氮添加使土壤微生物中各基因拷贝数分别下降40.97%(16S-rRNA)、78.38%(phoD)、67.92%(phoC)、74.37%(appA)。各基因拷贝数占总细菌基因拷贝数的比例显著下降了61%(phoD)、44%(phoC)、55%(appA)。土壤微生物量碳、微生物量磷含量与酸性磷酸单脂酶、碱性磷酸单脂酶、植酸酶活性及16S rRNA、phoD、phoC、appA基因丰度显著正相关。土壤铵态氮含量与酸性磷酸单脂酶、碱性磷酸单脂酶活性及16S rRNA、phoC、appA基因丰度显著负相关。酸性磷酸单酯酶活性与其基因丰度显著正相关,其他两种...     

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.     

不同添加方式植酸酶处理豆粕对牙鲆生长和饲料利用率的影响

以初始平均体重(2.02±0.02)g的牙鲆(Paralichthys olivaceus)为实验对象,进行为期70d的摄食生长实验,研究不同添加方式的植酸酶对牙鲆生长和饲料利用的影响。在5000.0g豆粕中添加2.5g植酸酶,然后用产朊假丝酵母(Candidautilis)进行发酵预处理,得到植酸酶预处理豆粕。共制作4种等氮等能(粗蛋白49.7%、总能20.9kJ/g)饲料,对照饲料主要以鱼粉为蛋白源;在对照饲料的基础上,用豆粕蛋白替代45%的鱼粉蛋白配制成豆粕组饲料;在每千克豆粕组饲料中添加1000IU植酸酶,配制成植酸酶组饲料;用植酸酶预处理豆粕蛋白替代45%的鱼粉蛋白配制成植酸酶预处理豆粕组饲料。结果表明,与对照组相比较,用豆粕蛋白替代饲料中45%的鱼粉蛋白,若不添加植酸酶则显著降低牙鲆的特定生长率(P0.01)、饲料效率、蛋白质效率和氮贮积率(P0.05);直接添加植酸酶组、植酸酶预处理豆粕组牙鲆的特定生长率、饲料效率、蛋白质效率和氮贮积率与鱼粉对照组相比较没有出现显著差异(P0.05);与不添加植酸酶的豆粕组相比较,在含豆粕饲料中添加1000IU/kg饲料的植酸酶显著提高牙鲆的特定生长率(P0.01)、氮贮积率(P0.05)和磷贮积率(P0.01),显著降低氮排放率(P0.05)和磷排放率(P0.01),但饲料效率和蛋白质效率没有显著变化(P0.05);在豆粕中添加植酸酶进行发酵预处理,降低了豆粕中植酸含量,在饲料中添加植酸酶预处理豆粕显著提高牙鲆的特定生长率(P0.01)、饲料效率、蛋白质效率和氮贮积率(P0.05),显著降低氮(P0.05)、磷和钙的排放率(P0.01)。       

通过参数相关性分析对发酵控制策略在植酸酶高密度发酵中的作用探讨

为了优化植酸酶高密度发酵条件,有必要获取在发酵过程中由于控制策略引起有关参数的实时变化及其关联性.本研究利用传感器对植酸酶工程菌高密度发酵过程进行数据在线采集,通过改变转速、接种量与补料甘油,探讨三方面控制因素对高密度发酵产酶过程参数具体影响及各参数变化之间的相关性,建立起与转速-细胞密度-溶氧-乳酸相关的发酵罐内外环...     

植酸酶对草鱼和新吉富罗非鱼消化酶活性的影响

植酸酶能水解植酸络合物,释放被植酸束缚的各种营养因子,因此能有效解除植酸与内源性消化酶的结合,促进消化酶的作用。本实验在全植物性饲料中添加植酸酶,研究其对草鱼(Ctenopharyngodon idellus)和新吉富罗非鱼(Oreochromis niloticus)淀粉酶及蛋白酶比活力的影响。以全植物性饲料为阴性对照组,添加磷酸氢钙(dibasic calcium phosphate,DCP)实验组为阳性对照组,另设4个不同梯度的植酸酶实验组(250 U/kg、500 U/kg、1 000 U/kg和2 000 U/kg)。实验选取健壮、规格齐整平均体质量为(12.59±0.09)g的草鱼和平均体质量为(9.59±0.12)g的新吉富罗非鱼,分别随机分为6个组,每组5个平行,每个平行20尾鱼。养殖8周后,草鱼平均体质量(18.29±0.63)g,新吉富罗非鱼平均体质量为(24.68±1.34)g,抽样取出胃、肠和肝胰脏用来分析淀粉酶和蛋白酶比活力。结果表明,植酸酶对无胃鱼草鱼和有胃鱼罗非鱼淀粉酶及蛋白酶比活力都有显著的促进作用。相比较而言,植酸酶对罗非鱼的应用效果较明显,低剂量就能显著提高其淀粉酶及蛋白酶比活力(P<0.05)。当植酸酶添加量达到1 000 U/kg时,草鱼和罗非鱼淀粉酶及蛋白酶比活力均达到峰值,此时,罗非鱼淀粉酶和蛋白酶比活力与阳性对照组无显著差异(P>0.05),而草鱼肝胰脏蛋白酶比活力显著高于阳性对照组(P<0.05)。植酸酶2 000 U/kg实验组,罗非鱼淀粉酶和蛋白酶比活力与1 000 U/kg植酸酶实验组无显著差异(P>0.05),但草鱼肝胰脏蛋白酶比活力显著低于1 000 U/kg植酸酶实验组(P<0.05)。因此,本实验条件下,植酸酶在草鱼和新吉富罗非鱼全植物性蛋白质配合饲料中的适宜添加量均为1 000 U/kg,生产实践中可通过添加植酸酶部分替代无机磷源。     

植酸酶的研究进展

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。植酸酶作为一种新型酶制剂,添加于食品和饲料中,能消除植酸引起的抗营养作用,提高蛋白质的生物利用率。本文综述有关植酸酶的分子结构、作用机理、生物学特征、基因结构的研究。     

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.     

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.