低嘌呤、高酶活纳豆芽孢杆菌液态发酵条件的优化

采用比浊法连续测定OD660nm值,绘制菌种生长曲线,研究纳豆芽孢杆菌种子生长条件,确定最佳接种时间为12h。并用单因素和正交设计实验的方法对纳豆激酶发酵条件进行优化,确定发酵条件的最优组合(g/L):全豆豆粉20,乳糖20,酵母浸膏10,KH2PO40.1,K2HPO4·3H2O 0.1,MgSO4·7H2O 0.5,NaCl 10,CaCl20.3,发酵液初期pH7.0,接种量2%,装液量30mL/50mL,摇速100r/min,发酵温度37℃,发酵时间36h。同时,优化后酶活由1005.77IU/mL提升到2835.50IU/mL,嘌呤总量由65.56mg/L降低到29.36mg/L,优化结果较为显著。

米曲霉与酱油曲霉高酶活菌株的诱变选育

酱油酿造过程中菌株所分泌的酶系直接影响原料的利用率和最终产品的品质。实验通过紫外诱变、亚硝基胍诱变以及紫外-亚硝基胍复合诱变方法分别对米曲霉和酱油曲霉菌株进行了诱变,采用酪蛋白透明圈法初筛以及福林酚法和DNS法复筛,根据碱性蛋白酶、中性蛋白酶、酸性蛋白酶、纤维素酶和α-淀粉酶酶活水平的变化,选育出数株米曲霉和酱油曲霉高酶活菌株。并对初筛方法的可行性和不同诱变方法对菌株改良的效果进行了探讨。

绿色动物食品——高酶活益生素在浚县问世

一种可增强动物抗病能力，防止病菌感染，同时又可提高饲料利用率，提高动物生长性能，提高动物肉类品质的绿色动物饲料添加剂——高酶活益生素日前在河南浚县立世集团研制成功，近日将投入批量生产。

高酶活益生素饲料添加剂

多年来人们在饲养畜禽动物时，为防病治病，总是用些抗生素或其它药物，抗生素和药物的应用，会给动物带来一定的负作用。如影响动物生长，降低动物生长性能，影响肉品质量等。肉品质量的下降将直接影响到人体健康。影响到肉类品的出口，欧美国家严禁抗生素和药物超标肉食品进口。

Aspergllus usamii B_1及B_(1—12)果胶酶性质、组分和应用的比较研究

B_(1—12)是由Asp·usamii B_1经NTG、UV、LiCl复合处理选育到的一株呈黑色的孢子是适于液体培养的高酶活果胶酶菌株。具有发酵周期短、易过滤等痔点,酒樁沉淀的酶制剂与B_1比较,酶作用的最适pH分别为4.0和4.5,最适温度均力50℃,但在30℃时,B_1仍保持较高酶活水平的稳定性。B_(1—12)果胶酶组分ex—PG酶活高于B_1 0.5—1倍左右,endo—PG和PE变化不明显.两者都含有糖化酶、纤维素酶、蛋白酶、花青素酶等非果胶酶组分,其中糖化酶活性比B_1下降一倍.应用于苹果醋、苹果汁、草莓汁、樱桃汁的澄清.B_(1—12)优于B_1,透光率可达90%以上,但对枣汁的澄清效果却次于B_1。

利用纤维素酶酶解粗饲料的研究——第四报:纤维素酶产生菌激光诱变育种

当前,纤维素酶在应用上要解决的主要问题之一是要获得活力高的纤维素酶产生菌菌种。国内外研究工作者利用诱变的方法获得了高酶活的菌株。美国陆军Natick实验室的Qm9419与原野生菌株Qm6a比较,酶活提高4倍;日本外山等诱变选育出高活力的黑曲霉T5;在国内中国科学院上海植生所采用物理化学诱变因素结合最后以高能电子处理获得了二株高酶活菌株EA3—867和N2—78,比其出发菌株分别提高酶活一倍和数倍。四川省生物所采用紫外线和亚硝酸复合处理。

Effect of ^(12)C^(6+) Ions Beam Irradiation on Seed Germination and Enzymes Activity in Seedlings of Sweet Sorghum

[ Objective] The aim was to study the effect of 12C6 ＋ ions beam irradiation to two varieties of sweet sorghum on seed germination and some enzymes activity in seedlings with different doses, and provided a theoretical foundation for sweet sorghum breeding. [ Method] After germination, the germination potential, germination fraction and enzyme activity were detected, respectively. [ Result] The results showed that with the dose increased, the germination potential of sweet sorghum increased first and then decreased, while their germination fraction presented ＂shoulder like shape＂ ; the activity of LDH, SOD, CAT and GSH-Px increased first and then decreased with doses, they presented slight differences among different enzymes. [ Conclusion] Low dose radiation could accelerate germination of sweet sorghum seeds and enzyme activity could remain at a relatively high level. Enzyme activity decreased with high doses and the growth of sweet sorghum was inhibited.

Advances in Melatonin and Its Functions in Plants

Abstract Melatonin （N-acetyi-5-methoxytryptamine） is a well-known animal hormone, which is synthesized and secreted by pineal gland and takes part in the regulation of circadian rhythm in animals. At present it has been observed that melatonin is widely existed in higher plants while there have no enough studies on functions of melatonin in plants. Researches have already indicated that the possible functions of melatonin in plants include regulating photoperiod, participating in growth regulation, clearing active oxygen, and promoting activity of antioxidase. General reviews upon functions of melatonin in plants are made upon experiments in recent years. We fo-cus on the demonstrated and predicted biological functions of melatonin in plants to bring researchers up to date on this field. The weakness in present studies and the main research directions are also pointed out.

Inducing Expression and Reaction Characteristic of Nitrile Hydratase from Rhodococcus sp. SHZ-1

Inducing expression and the reaction characteristic of nitrile hydratase （NHase） from Rhodococcus sp. SHZ-1 were investigated. The results showed that the expression of NHase was greatly enhanced with the cooperation of acrylonitrile and ammonium chloride as inducer in the medium and the specific activity of NHase was increased of 44%. Then the temperature, pH, concentration of acrylonitrile and acrylamide were evaluated, which affected the activity and reaction characteristic of NHase. It was found that the temperature and concentration of acrylarnide were the most important factors for the catalyzation of NHase. The optimal catalysis temperature of NHase from Rhodococcus sp. SHZ-1 was 30℃, and the activation energy of the hydration of NHase was 90.2kJ·mol^-1 in the temperature range from 5℃ to 30℃. Kmof NHase was 0.095mol·L^-1 using acrylonitrile（AN） as substrate, and NHase activity was inhibited seriously when acrylonitrile concentration was up to 40g·L^-1, the substrate inhibition constant Ki is 0.283mol·L^-1. Moreover, the NHase from Rhodococcus sp. SHZ-1 had very strong tolerance to acrylamide, in which the final concentration of acrylamide reached to 642g·L^-1 and the residual activity of NHase still maintained 8.6% of the initial enzyme activity.

Soil Phosphorus Composition and Phosphatase Activities along Altitudes of Alpine Tundra in Changbai Mountains,China

Alpine tundra ecosystems have specific vegetation and environmental conditions that may affect soil phosphorus （P） composition and phosphatase activities. However, these effects are poody understood. This study used NaOH-EDTA extraction and solution ^31P nuclear magnetic resonance （NMR） spectroscopy to determine soil P composition and phosphatase activities, including acid phosphomonoesterase （AcP）, phosphodiesterase （PD） and inorganic pyrophosphatase （IPP）, in the alpine tundra of the Changbai Mountains at seven different altitudinal gradients （i.e., 2000 m, 2100 m, 2200 m, 2300 m, 2400 m, 2500 m, and 2600 m）. The results show that total P （TP）, organic P （OP）, OP/TP, NaOH-EDTA extracted P and AcP, PD, and IPP activities over the altitude range of 2500-2600 m are significantly lower than those below 2400 m. The dominant extracted form of P is OP （73%0-83%） with a large proportion of monoesters （65%0-72%）, whereas inorganic P is present in lower proportions （17%-27%）. The activity of AcP is significantly positively correlated with the contents of soil OP, total carbon （TC）, total nitrogen （TN）, and TP （P 〈 0.05）, indicating that the AcP is a more sensitive index for responding P nutrient storage than PD and IPP. Soil properties, P composition, and phosphatase activities decrease with increased altitude and soil pH. Our results indicate that the distribution of soil P composition and phosphatase activities along altitude and AcP may play an important role in P hydrolysis as well as have the potential to be an indicator of soil quality.

Deletion of a Non-Catalytic Region Increases the Enzymatic Activity of a β-Agarase from Flammeovirga sp. MY04

A Glycoside hydrolase （GH） typically contains one catalytic module and varied non-catalytic regions （NCRs）. However, effects of the NCRs to the catalytic modules remain mostly unclear except the carbohydrate-binding modules （CBMs）. AgaG4 is a GH16 endo-β-agarase of the agarolytic marine bacterium Flammeovirga sp. MY04. The enzyme consists of an extra sugar-binding peptide within the catalytic module, with no predictable CBMs but function-unknown sequences in the NCR, which is a new characteristic of agarase sequences. In this study, we deleted the NCR sequence, a 140-amino acid peptide at the C-terminus and expressed the truncated gene, agaG4-T140, in Escherichia coli. After purification and refolding, the trtmcated agarase rAgaG4-T140 retained the same catalytic temperature and pH value as rAgaG4. Using combined fluorescent labeling, HPLC and MS/MS techniques, we identified the end-products of agarose degradation by rAgaG4-T140 as neoagarotetraose and neoagarohexaose, with a final molar ratio of 1.53：1 and a conversion ratio of approximately 70%, which were similar to those of rAgaG4. However, the truncated agarase rAgaG4-T140 markedly decreased in protein solubility by 15 times and increased in enzymatic activities by 35 times. The oligosaccharide production of rAgaG4-T140 was approximately 25 times the weight of that produced by equimolar rAgaG4. This study provides some insights into the influences of NCR on the biochemical characteristics of agarase AgaG4 and implies some new strategies to improve the properties of a GH enzyme.

The difference of soluble sugar contents and starch synthetic key enzyme activities between high and low starch cultivar of cassava

The difference of soluble sugar contents and starch synthetic key enzyme activities between high starch cultivar and low starch cultivar were studied in this experiment, of which FuXuan01, GR891, SC124, and SC201 were used as materials. The results showed that the contents of reducing sugar in root tube of low starch cultivars were higher than those of high starch cultivars in all growth periods, the contents of sucrose in root tube of high starch cultivar were higher than those of low starch cultivars in the early growth period, but, lower in the late growth period, and there were almost no difference in the contents of soluble sugar in root tube between high starch cultivar and low starch cultivar in the early growth period, but notable difference in the late growth period and the soluble sugar contents of low starch cultivars were higher than those of high starch cultivars. It also-showed that the activities of ADPGPpase, SSS, and SBE （starch branching enzyme） of high starch cultivars were higher than those of low starch cultivars. It was evident that there were close correlations between the content of sucrose, reducing sugar and soluble sugar in root tube, and the activities of ADPGPpase, SSS, SBE, and the starch accumulation in the root tubers of cassava. It was quite evident in this experiment that the soluble sugar and starch synthetic key enzyme were main factors controlling starch accumulation in root tubers. These results provided important indication for physiological controlling in high starch cultivation and gene engineering breeding of high starch cultivar of cassava.

Effects of Iron and Aluminum Oxides and Kaolinite on Adsorption and Activities of Invertase *1

Experiments were conducted to study the influences of synthetic bayerite, non crystalline aluminum oxide (N AlOH), goethite, non crystalline iron oxide (N FeOH) and kaolinite on the adsorption, activity, kinetics and thermal stability of invertase. Adsorption of invertase on iron, aluminum oxides fitted Langmuir equation. The amount of invertase held on the minerals followed the sequence kaolinite > goethite > N AlOH > bayerite > N FeOH. No correlation was found between enzyme adsorption and the specific surface area of minerals examined. The differences in the surface structure of minerals and the arrangement of enzymatic molecules on mineral surfaces led to the different capacities of minerals for enzyme adsorption. The adsorption of invertase on bayerite, N AlOH, goethite, N FeOH and kaolinite was differently affected by pH. The order for the activity of invertase adsorbed on minerals was N FeOH > N AlOH > bayerite > reak goethite > kaolinite. The inhibition effect of minerals on enzyme activity was kaolinite > crystalline oxides > non crystalline oxides. The pH optimum of iron oxide and aluminum oxide invertase complexes was similar to that of free enzyme (pH 4.0), whereas the pH optimum of kaolinite inv ertase complex was one pH unit higher than that of free enzyme. The affinity to substrate and the maximum reaction velocity as well as the thermal stability of combined invertase were lower than those of the free enzyme.

Effect of Bu-Zhong-Yi-Qi-Tang on deficiency of N-glycan/nitric oxide and islet damage induced by streptozotocin in diabetic rats

AIM: To investigate the effect of Bu-Zhong-Yi-Qi-Tang (Decoction for Reinforcing Middle Jiao and Replenishing Qi) on deficiency of N-glycan/nitric oxide (NO) and islet damage induced by injecting two medium doses of streptozotocin (STZ). METHODS: Diabetes was induced by intraperitoneal injection of STZ at 55 mg/kg on day 1 and day 8. Islet damage was evaluated using a scoring system. Nitrite, nitrate, α-mannosidase and amylase activities were measured by colorimetry. N-glycan patterns of amylase were determined with lectin [ConA, pisum sativum agglutinin (PSA), peanut agglutinin (PNA), and lens culinaris agglutinin (LCA)] affinity precipitation method. RESULTS: Severe islet necrosis and mild islet atrophy were observed in diabetic rats. The number and size ofislets, the activities of α-mannosidase, amylase and nitrite were decreased, while the binding of PNA and LCA to amylase was increased. All of which were improved after treatment with Bu-Zhong-Yi-Qi-Tang. Islet damage was significantly correlated with nitrite, nitrate, α-mannosidase, amylase and the binding of LCA, PNA, and PSA to amylase.CONCLUSION: STZ-induced islet damage is related to N-glycan deficiency in proteins by blocking α-mannosidase activity and no deficiency, accumulation of unfolded proteins, and endoplasmic reticulum stress and activation of cellular signals, all of which are improved after treatment with Bu-Zhong-YiQi-Tang.

Soil Cellulase Activity and Fungal Community Responses to Wetland Degradation in the Zoige Plateau, China

Four soil types(peat, marsh, meadow, and sandy) in the Zoige Plateau of China are associated with the severity of wetland degradation. The effects of wetland degradation on the structure and abundance of fungal communities and cellulase activity were assessed in these 4 soil types at 3 depths using DGGE(Denatured Gradient Gel Electrophoresis), q PCR(Quantitative Real-time PCR),and 3,5-dinitrosalicylic acid assays. Cellulase activity and abundance of the fungal community declined in parallel to the level of wetland degradation(from least to most disturbed). DGGE analysis indicated a major shift in composition of fungal communities among the4 soil types consistent with the level of degradation.Water content(WC), organic carbon(OC), total nitrogen(TN), total phosphorus(TP), available nitrogen(AN), and available phosphorus(AP) were strongly correlated with cellulase activity and the structure and abundance of the fungal community.The results indicate that soil physicochemical properties(WC, OC, TN, TP, AN, and AP), cellulase activity, and diversity and abundance of fungal communities are sensitive indicators of the relative level of wetland degradation. WC was the major factorinvolved in Zoige wetland degradation and lower WC levels contributed to declines in the abundance and diversity of the fungal community and reduction in cellulase activity.

Antioxidant Defense System in Wheat （Triticum aestivum L.） Seedlings under Heat Stress and Revival Conditions

The present investigation was carried out to investigate the effect of heat stress and revival on some antioxidative enzymes and metabolites in leaves of the wheat （Triticum aestivum L.） seedlings of heat susceptible （cv. WH 147 and HS 277） and heat tolerant （cv. WH 1021 and HW 2045） cultivars. Seven days old seedlings grown at 25 ℃ were exposed to 40 ℃ for 6 h and these seedlings were again brought to 25 ℃. The observations were recorded in the leaves of control, stressed and revived seedlings on 2nd and 4th day of revival. For the selection ofthermo-tolerant cultivars, screening of the thirty-six cultivars was done based on wilting of primary leaf and values of chlorophyll fluorescence. The MDA （malondialdehyde） and H2O2 concentration in leaves of wheat seedlings increased at the high temperature. There was enhancement in the activities of antioxidative enzymes, viz. CAT （catalase）, POX （peroxidase）, GR （glutathione reductase） and APX （ascorbate peroxidase） in leaves of the tolerant and susceptible cultivars under heat stress, however, higher percent increase was observed in tolerant cultivars. Heat stress increased the SOD （superoxide dismutase） activity in tolerant cultivars but activity declined in susceptible cultivars. On revival, the activities of the CAT, POX and GR declined in comparison to stressed seedlings but remained higher as compared to control. Ascorbate peroxidase activity remained higher on 2nd day and 4th day of revival in all the cultivars.

THE INCREASE IN PLASMINOGEN ACTIVATOR INHIBITOR TYPE-1 EXPRESSION BY STIMULATION OF ACTIVATORS FOR PEROXISOME PROLIFERATOR-ACTIVATED RECEPTORS IN HUMAN ENDOTHELIAL CELLS

Objective.To investigate the effect of peroxis ome proliferator-activated recept ors(PPARs )activators on plasminogen activator inhibitor ty pe-1(PAI-1)expression in human umbilical vein e ndothelial cells and the possi-ble mechanism.Methods.Human umbilical vein endothelial ce lls(HUVECs )were obtained from normal fetus,and cul-tured conventionally.Then the HUVECs were exposed to test agents(linolenic acid,linoleic acid,oleic acid,stearic acid and prostaglandin J 2 respectively)in varying concentrations with fresh media.RT -PCR and ELISA were applied to determine the expression of PPARs and PAI-1in HUVECs.Results.PPARα,PPARδand PPARγmRNA were detected by using RT-PCR in HUVECs.Treatment of HUVECs with PPARαand PPARγactivators---linolenic acid,linoleic acid,oleic acid and prostaglandin J 2 respectively,but not with stearic a cid could augment PAI-I mRNA expression and protein secretion in a concentration-dependent manner.However,the mRNA expressions of 3subclasses of PPAR with their activators in HUVECs were not changed compared w ith controls.Conclusion.HUVECs express PPARs.PPARs activators may increase PAI-1expression in ECs,but the underlying mechanism remains uncle ar.Although PPARs expression was not enhanced after stimulated by their activators in ECs,the role of functionally active PPARs in regulating PA I-1expression in ECs needs to be further investigated by using transient gen e transfection assay.

Effects of elevated CO_2 on the nutrient compositions and enzymes activities of Nilaparvata lugens nymphs fed on rice plants

Elevated CO 2 may reduce the tolerance of Nilaparvata lugen (N. lugens) to adverse environmental factors through the biological and physiological degeneration of N. lugens. In an artificial climate box, under 375 and 750 μL L 1 CO 2 levels, the rice stems nutrient content, the nutrient content and enzyme activities of N. lugens nymph fed on rice seedlings exposed to ambient and elevated CO 2 were studied. The results showed that rice stems had significantly higher protein and total amino acid levels under ambient than elevated CO 2 levels. Nymphs had significantly higher protein levels in the ambient CO 2 treatment, while their glucose levels were significantly lower under ambient CO 2 conditions. Significantly higher trypsin activity was observed in nymphs grown in elevated CO 2 . Significantly lower activities of the protective enzymes total superoxide dismutase and catalase were observed in the nymphs under ambient CO 2 . Meanwhile, the activity of the detoxification enzyme glutathione S-transferase was significantly higher in the ambient CO 2 treatment. Measuring how energy and resources were allocated to enzymes in N. lugens nymphs under elevated CO 2 conditions can provide a more meaningful evaluation of their metabolic tolerances to adverse climatic conditions.