海洋芽孢杆菌N11-8产蛋白酶的发酵条件优化

本研究采用响应面法(RSM)对南极海洋芽孢杆菌(Bacillus sp.)N11-8液体发酵产蛋白酶PBN11-8的发酵条件进行快速优化。通过单因素实验初步确定南极海洋芽孢杆菌N11-8产蛋白酶的最佳碳源和氮源分别为可溶性淀粉和蛋白胨；并通过Plackett-Burman(PB)设计对影响其产酶相关因素进行评估，筛选出具有显著效应的3个因素：温度、氮源和碳源；以最陡爬坡实验逼近至上述因子最大响应区域，进而采用RSM法对其最佳水平范围进行研究，确定最优发酵条件为：可溶性淀粉3 g/L，蛋白胨13.1 g/L，酵母浸粉2.9 g/L，NaCl 5 g/L，KH2PO4 1 g/L，FeCl3·6H2O 2 mmol/L，初始pH值为7.0，温度为34.0℃，转速为200 r/min，装液量为50 ml/250 ml，接种量为4%，培养时间为60 h。最终优化后的酶活达到90.5 U/ml，比初始酶活提高了23.6%。     

An improved method for extraction of sorghum polymeric protein complexes

A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm.     

Dynamics of upward and downward N2O and CO2 fluxes in ploughed or no-tilled soils in relation to water-filled pore space, compaction and crop presence

Sharp peaks in nitrous oxide (N₂O) fluxes under no-tillage in wet conditions appear to be related to near surface soil and crop cover conditions. Here we explored some of the factors influencing tillage effects on short-term variations in gas flux so that we could learn about the mechanisms involved. Field investigations revealed that a cumulative emission of 13 kg N₂O–N ha⁻¹ over a 12-week period was possible under no-tillage for spring barley. We investigated how reducing crop cover and changing the structural arrangement of the water-filled pore space (WFPS) by short-term laboratory compaction influenced N₂O and carbon dioxide (CO₂) fluxes in upward and downward directions in core samples from tilled and untilled soil. Increasing the downward flux of N₂O within a soil profile by changing soil or moisture conditions may increase the likelihood of its further reduction to N₂ or dissolution. We took undisturbed cores from 3 to 8 cm depth, equilibrated them to −1 or −6 kPa matric potential, incubated them and measured N₂O and CO₂ fluxes from the upper and lower surfaces in a purpose-designed apparatus before and after compaction in an uniaxial tester. We also measured WFPS, air permeability, bulk density and air-filled porosity before and after compaction. Spring barley was tested in 1999 and winter barley in 2000.Fluxes of N₂O were from 1.5 to 35 times higher from no-tilled than ploughed even where the soil was of similar bulk density. Reduction of the crop cover increased CO₂ flux and could reduce N₂O flux. The effects of structural changes induced by laboratory compaction on the fluxes of N₂O and CO₂ were not influenced greatly by the tillage and crop cover treatments. Fluxes from the upper surfaces of cores (corresponding to 3 cm soil depth, upwards direction) could be up to 100 times greater (N₂O) or 8 times (CO₂) than from the lower surfaces (8 cm depth, downwards direction). These differences between surfaces were greatest when N₂O fluxes were very high in no-tilled soil (4.2 mg N₂O–N m⁻² h⁻¹) as occurred when WFPS exceeded 80% or became blocked with water, an effect that was increased by our compaction treatment. In general N₂O fluxes increased with WFPS. The production and emission of N₂O were strongly influenced by the soil physical environment, the magnitude of the water-filled pore space and continuity of the air-filled pore space in particular, produced in no-till versus plough cultivation.     

团头鲂肠道肝胰脏消化酶的初步研究

本文采用动力学方法测定了团头鲂肝胰脏淀粉酶和肠道蛋白酶、淀粉酶的活性分布及pH值对各消化酶活性的影响,结果表明:食糜中的蛋白酶、淀粉酶活性沿消化道从前至后逐渐减弱,而肠黏膜中各消化酶活性则基本上是由前到后逐渐增加。在pH值6.0～9.0范围内,肠道蛋白酶活性在pH为8.6时为最大,在pH值6.2～8.6范围内,肝胰脏和肠道淀粉酶活性在pH值7.8时为最大。     

内蒙古根河14种野生大型经济真菌生物活性测定

对内蒙古根河市14种野生大型经济真菌的抗菌活性及凝集素、蛋白酶、磷酸酶等生物活性进行了测定。结果表明在被测的14个样品中,具有蛋白酶活性的有4种、具有凝集素活性的有7种、具有抗植物病原真菌活性的有3种,开发和利用这些野生大型经济真菌将取得巨大的经济效益和社会效益。     

A review of cyanobacterial odorous and bioactive metabolites: Impacts and management alternatives in aquaculture

Increased demand has pushed extensive aquaculture towards intensively operated production systems, commonly resulting in eutrophic conditions and cyanobacterial blooms. This review summarizes those cyanobacterial secondary metabolites that can cause undesirable tastes and odors (odorous metabolites) or are biochemically active (bioactive metabolites) in marine and freshwater, extensive and intensive aquaculture systems. For the scope of this paper, biochemically active metabolites include (1) toxins that can cause mortality in aquaculture organisms or have the potential to harm consumers via accumulation in the product (hepatotoxins, cytotoxins, neurotoxins, dermatoxins, and brine shrimp/molluskal toxins), (2) metabolites that may degrade the nutritional status of aquaculture species (inhibitors of proteases and grazer deterrents) or (3) metabolites that have the potential to negatively affect the general health of aquaculture species or aquaculture laborers (dermatoxins, irritant toxins, hepatotoxins, cytotoxins). Suggestions are made as to future management practices in intensive and extensive aquaculture and the potential exposure pathways to aquaculture species and human consumers are identified.     

芽孢杆菌的筛选及其在豆粕发酵中的应用

筛选出耐酸耐胆盐且产蛋白酶、淀粉酶性能较好的优势芽孢杆菌菌株,以期制备高效复合益生素或应用于豆粕发酵。首先,对分离并镜检出的17株菌株进行耐酸耐胆盐特性初筛,而后对筛出的3株菌株进行耐酸耐胆盐梯度复筛,随后检测其发酵豆粕的蛋白酶和淀粉酶活性以及对抗原蛋白的降解能力。最后通过16SrRNA扩增比对鉴定筛选的菌种,进而通过MEGA4.0构建系统进化树进行分析。成功筛选出1株耐酸耐胆盐、产蛋白酶和淀粉酶,且能高效降解豆粕抗原蛋白的解淀粉芽孢杆菌B9。     

Optimal dietary protein requirement for juvenile ivory shell, Babylonia areolate

A growth experiment was conducted to determine the optimal dietary protein requirement for juvenile ivory shell reared in indoor aerated aquaria. Six isoenergetic experimental diets using fish meal, casein and gelatin as protein sources were formulated to contain graded levels of protein (27, 33, 38, 43, 49 and 54% of dry diet, respectively). Triplicate groups of 40 shells (average weight 93.50 ± 1.70 mg) were stocked in 120-l tanks and fed to apparent satiation twice daily for 8 weeks. The results showed that the growth performance and feed utilization were significantly affected by dietary protein level (P < 0.05). Maximum weight gain, mean protein gain, specific growth rate and soft body to shell ratio occurred at 43% dietary protein level (P < 0.05). There were significant differences in protein, lipid, moisture and ash content in soft body; except that ash content in shell was not significantly affected by dietary protein level. Pepsin activity in soft body tissue significantly increased with dietary protein level up to 43%, and trypsin-like enzyme activity increased with dietary protein level up to 49%. However, lipase activity in soft body decreased with increasing dietary protein level. However, no significant differences (P < 0.05) in survival, calcium, phosphorus concentration in the shell and soft body were found among dietary treatments. Quadratic regression analysis of weight gain against dietary protein level indicated that the optimal dietary protein requirement for maximum growth and feed utilization of juvenile ivory shell is 45% of dry diet.     

不同生长阶段斑节对虾消化酶活性变化

通过对斑节对虾不同生长阶段蛋白酶、脂肪酶及淀粉酶活性变化的分析,探讨了斑节对虾在生长过程中对蛋白质、脂肪和碳水化合物的消化利用。结果表明,根据消化酶活性,可将斑节对虾生长分为两个阶段。第1阶段,在体长10～12.5 cm阶段之前,随着个体的增大蛋白酶和脂肪酶活性不断升高;第2阶段,在12.5 cm之后,蛋白酶和脂肪酶活性呈现出一定程度的下降。肝胰腺、胃、肠3种不同消化器官的消化酶活性在不同生长期也不一致,大致上蛋白酶活性从高到低依次为:肝胰腺>胃>肠;脂肪酶活性:肠>肝胰腺>胃;淀粉酶活性:肠>胃>肝胰腺。     

Inhibition of protease activities in discus Symphysodon spp. by three plant meals

Based on biochemical assays and electrophoretical methods, the inhibitory effects of three plant meals (soybean meal, wheat meal, winged bean meal) on digestive alkaline proteases of discus were investigated. Casein assays revealed that increasing levels of soybean meal caused a linear inhibitory effect on activity of protease. SDS-PAGE images revealed that trypsin and chymotrypsin were the inhibited enzymes. Soybean showed the lowest inhibition rate followed by wheat meal and raw winged bean. There was a quadratic relationship between wheat meal levels and its inhibition of protease acitivity. The highest inhibitory effect was obtained with the winged bean meal with inhibition of caseinolytic activities ranging from 3.6–98.6%. Results from this study showed the potential of both soybean meal and wheat meal as ingredients for practical diet for discus, while demonstrating the need for further improvement in processing method for winged bean meal.