Characterization of a thermostable extracellular beta-glucosidase with activities of exoglucanase and transglycosylation from Paecilomyces thermophila

The purification and characterization of a novel extracellular beta-glucosidase from Paecilomyces thermophila J18 was studied. The beta-glucosidase was purified to 105-fold apparent homogeneity with a recovery yield of 21.7% by DEAE 52 and Sephacryl S-200 chromatographies. Its molecular masses were 116 and 197 kDa when detected by SDS-PAGE and gel filtration, respectively. It was a homodimeric glycoprotein with a carbohydrate content of 82.3%. The purified enzyme exhibited an optimal activity at 75 degrees C and pH 6.2. It was stable up to 65 degrees C and in the pH range of 5.0-8.5. The enzyme exhibited a broad substrate specificity and significantly hydrolyzed p-nitrophenyl-beta- d-glucopyranoside ( pNPG), cellobiose, gentiobiose, sophorose, amygdalin, salicin, daidzin, and genistin. Moreover, it displayed substantial activity on beta-glucans such as laminarin and lichenan, indicating that the enzyme has some exoglucanase activity. The rate of glucose released by the purified enzyme from cellooligosaccharides with a degree of polymerization (DP) ranging between 2 and 5 decreased with increasing chain length. Glucose and glucono-delta-lactone inhibited the beta-glucosidase competitively with Ki values of 73 and 0.49 mM, respectively. The beta-glucosidase hydrolyzed pNPG, cellobiose, gentiobiose, sophorose, salicin, and amygdalin, exhibiting apparent Km values of 0.26, 0.65, 0.77, 1.06, 1.39, and 1.45 mM, respectively. Besides, the enzyme showed transglycosylation activity, producing oligosaccharides with higher DP than the substrates when cellooligosaccharides were hydrolyzed. These properties make this beta-glucosidase useful for various biotechnological applications.     

Changes in organic carbon content and its physical and chemical distribution in paddy soils cultivated under different fertilisation practices

Purpose

Chemical protection facilitates soil organic carbon (SOC) sequestration and stabilisation due to a strong chemical binding with mineral surfaces and metal ions (e.g. iron [Fe], aluminium [Al] and calcium [Ca]). However, there is not much information regarding the role of chemical protection in SOC stabilisation in paddy soils, particularly in terms of the specific forms of organo-mineral complexes such as Fe-, Al- and Ca-bonded OC.

Materials and methods

We sampled paddy soils at the 0–20 cm soil layer from a long-term field experiment (initiated in 1981) conducted under humid subtropical conditions in China, which has five fertilisation treatments (i.e. control treatment without fertiliser [CK], chemical fertiliser only [CF], green manure [GM], Straw and Manure) with equivalent nutrient inputs (i.e. N, P₂O₅ and K₂O at the rates of 135–67.5–135 kg ha^?1, respectively, for both early and late rice) except CK. We determined the chemical binding forms of SOC and the associated soil properties in the particulate fraction (PF, >53 μm) and the mineral-associated fraction (MAF, <53 μm), which were obtained using a low-energy ultrasonic dispersion procedure, of a paddy soil in the long-term fertilisation experiment.

Results and discussion

Iron- and Al-bonded OC (Fe/Al-OC) was the dominant fraction and made up 55–70% of the total SOC in the paddy soil, while Ca-bonded OC (Ca-OC) was only a minor fraction (<4%). The Fe/Al-OC was mainly allocated in the MAF (52–67%), indicating that the chemical protection of SOC occurred mostly in the finer particle fractions. Long-term application of organic amendments increased the contents of bulk SOC by 27–34% (P < 0.05), of Fe/Al-OC by 9–16% and of Ca-OC by 35–83% (P < 0.05), whereas the sole application of chemical fertiliser had no significant effects on SOC contents of the paddy soil compared with the treatment without fertiliser inputs. Both amorphous Fe and Al extracted by ammonium oxalate (Fe_ox and Al_ox) showed significant correlations with Fe/Al-OC (r = 0.52 and 0.78, respectively), but Al_ox appeared to have a greater influence on C stabilisation in the paddy soil.

Conclusions

These results demonstrated that the dominant chemical binding forms of SOC in the paddy soils were Fe/Al-OC and amorphous Fe/Al oxyhydrates, especially amorphous Al, contributed mostly to the chemical stabilisation of SOC.

     

社会主义初级阶段和阶段性特征是我们谋改革、求发展的立足点

改革开放近30年来,我国取得了举世瞩目的成就,但是人口多、基础差、人民群众尚不富裕、发展不平衡的基本国情没有根本改变,这就要求我们必须清醒地认识我国仍将处于并将长期处于社会主义初级阶段这一基本国情;同时,也要清醒认识我国发展的阶段性特征。深刻认识国情和所处的历史阶段,是正确提出和贯彻党的理论和路线、方针、政策的基本依据,也是推进各项事业发展的重要前提。当前我们贯彻落实科学发展观、深化改革开放的基本立足点就是社会主义初级阶段和阶段性特征。     

Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.     

多效唑对大蒜试管微鳞茎形成和膨大的影响

以MS为基本培养基,用地方品种阿城紫皮大蒜茎尖为外植体进行离体培养试验。结果表明:在一定范围内,随着多效唑浓度的增加,抑制鳞茎的形成与分化,鳞茎发生数减少,地上部生长受抑制严重,叶片和根系变短增粗。添加6 mg/L多效唑为诱导鳞茎形成和促进膨大的最佳浓度,鳞茎形成率109.00%,鳞茎鲜重为307.07 mg,鳞茎指数1.15。     

DNA Immunization Against Very Virulent Infectious Bursal Disease Virus with VP2‐4‐3 Gene and Chicken IL‐6 Gene

The present study was to investigate the feasibility and efficiency of the DNA vaccine to protect chickens against very virulent infectious bursal disease virus (vvIBDV) infection. A plasmid DNA carrying VP2‐4‐3 genes of vvIBDV SH95 and a plasmid DNA carrying chicken interleukin‐6 (ChIL‐6) genes were constructed and designated as pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 respectively. Several DNA vaccination experiments were performed: 1‐week‐old chickens were intramuscularly injected with only plasmid pcDNA3‐VP2, pALTER‐MAX‐VP2‐4‐3 or mixture with pALTER‐MAX‐ChIL‐6. The chickens at 4 weeks old were orally inoculated with vvIBDV SH95. The results showed that immunization with the mixture of pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 three times conferred protection for 90% of chickens. Enzyme‐linked immunosorbent assay (ELISA) antibody titres in chickens immunized together with pALTER‐MAX‐ChIL‐6 were higher than those immunized simply with plasmid pcDNA3‐VP2 or pALTER‐MAX‐VP2‐4‐3. IBDV was not detected in the bursa of the protected chickens at 8 days after challenge by RT‐PCR. The results indicate that protection against vvIBDV can be achieved by using the VP2‐4‐3 gene of vvIBDV as a DNA vaccine. Furthermore, the simultaneous injection of ChIL‐6 plasmid significantly increased the protection after challenge with the very virulent strain.     

遮荫对水曲柳幼苗根系衰老的影响

用温室盆栽方法对自然光照和遮荫处理下水曲柳幼苗根系可溶性蛋白、MDA含量和DNA片断化进行了研究.结果表明:遮光幼苗的根系中可溶性蛋白含量高于正常的苗木;不同处理幼苗粗根中MDA浓度差异显著(p=0.009),遮荫处理苗木要低于正常苗木;DNA琼脂糖凝胶电泳没有明显的条带出现,说明短时间遮荫对水曲柳根系衰老有一定影响.     

除草剂条件下三裂叶豚草竞争对大豆生长发育的影响

在有无除草剂的影响下,研究不同密度三裂叶豚草(0、0.08、0.32株·m-2)对大豆生长发育的影响,探究三裂叶豚草对大田旱生作物的危害表现.结果表明:未施除草剂条件下大豆的株高要大于施除草剂条件下大豆株高;施除草剂条件下,苗期大豆分枝数随三裂叶豚草密度的增加逐渐增加,在结荚期,则随豚草密度的增加而呈先增加后降低的趋势;大豆茎重在开花期和结荚期随着三裂叶豚草密度的增加而降低,大豆地下部分生物量,除开花期施除草剂的处理外,均随着三裂叶豚草密度的增加而呈先增加后减少的趋势.因此可通过向农田中适当的施用除草剂抑制三裂叶豚草对农作物的影响.     

龙眼压差通风预冷装置风速控制与能耗分析

针对龙眼的物理特性,设计了一套龙眼压差通风预冷试验装置.根据果蔬压差预冷7/8时间特性,通过试验得到了龙眼包装箱压差与风速关系以及异步电动机频率与风速关系.进一步研究龙眼压差预冷装置能耗与风速关系可知,对于龙眼果实,较小的风速预冷能耗较小,预冷时间较长;当预冷时间按要求设定时,在风机允许范围内控制风速按一定规律递增,直到龙眼终止温度,可以在设定时间内完成龙眼预冷,且能耗最小.