Clone and expression of β-1,3-1,4-glucanase in Pichia pastoris

β-1,3-1,4-d-葡聚糖酶是一类专一性降解β-1,3-1,4-葡糖苷键中的β-1,4-糖苷键,产生小分子还原糖的水解酶,广泛应用于啤酒工业和饲料工业中。本研究根据毕赤酵母密码子偏好性优化β-1,3-1,4-葡聚糖酶基因序列,采用PCr法将其插入毕赤酵母表达载体PPICzαA,经SACI线性化后电击整合入毕赤酵母X-33基因组,构建重组酵母;经菌落PCr验证和摇瓶筛选,获得一株X-33/PPICzαA-bgl,甲醇诱导96H后,酶活力达308.5u/Ml,经SdS-PAgE电泳,实际蛋白分子量约为33ku。β-1,3-1,4-d-葡聚糖酶最适反应PH为5.0,最适反应温度为50℃。β-1,3-1,4-glucanase,which can hydrolyzeβ-1,3-1,4-glucan by cutting off theβ-1,4-glycosidic linkages and release shorter reducing sugar,has been applied in the brewing and animal feed additive industry.It can effectively improve digestibility of barley-based diets and reduce enteritis.In this report,the gene ofβ-1,3-1,4-glucanase,which was optimized according to Pichia pastoris condon usage,was amplified by polymerase chain reaction and ligated into the expression vector pPICZαA.The recombinant vector was lineared by Sac I and transformed into Pichia pastoris X-33 by electroporation.Then the optimal positive transformant,named X-33/pPICZαA-bgl,was screened by colony PCR and shake flask fermentation.The enzyme activity of recombinantβ-1,3-1,4-glucanase couldreach 308.5U/mL after 96h methanol induction.Results of SDS-PAGE showed that about 33ku protein of theβ-1,3-1,4-glucanase was expressed.Acidity and temperature optimal for this recombinant enzyme was pH5.0 and 50℃,respectively.福建省农业科学院青年基金(2012DQC-11); 公益性行业(农业)科研专项(201303094-05

Clavicoronol,a new metabolite,from the mycelia culture of Clavicorona pyxidata

目的对珊瑚菌(ClAVICOrOnA PyXIdATA)菌体培养物的活性成分进行分离。方法用多种色谱技术对化合物进行分离纯化,并用光谱技术和单晶X射线衍射技术鉴定化合物的结构。结果从中分离到2个化合物:ClAVICOrOnOl(1)、腺苷(2)。结论化合物1为新化合物,并通过单晶X射线衍射技术确定了2的立体构型。Aim To explore the new bioactive metabolites from the mycelia culture of the edible mushroom Clavicorona pyxidata.Methods The constituents were isolated and purified by column chromatography,and their structures were identified by spectroscopic analyses,including 1D-and 2D-NMR data,and single-crystal X-ray diffraction.Results A new natural product,named clavicoronol(1) along with adenosine(2),were obtained.Conclusion Compound 1 is a new compound and stereochemistry of 2 was confirmed by a single-crystal X-ray diffraction.ThisworkwasfinanciallysupportedbytheNationalNaturalScienceFoundationofChina(30500632);theNationalScienceFundforDistinguishedYoungScholarstoY.-M.Shen(30325044

A TWO-PARTICLE TURBIDIMETRIC LATEX IMMUNOASSAY FOR THE DETECTION OF SPECIFIC ANTIBODIES OF TRICHINELLA SPIRALIS

目的　建立旋毛虫病的快速检测技术。方法　基因工程抗原包被有色乳胶颗粒，抗抗体包被磁性颗粒，在抗体存在下形成抗原－ 乳胶－ 抗体－ 抗抗体－ 磁性颗粒的复合场，在磁场作用下沉淀下来，从而达到快速检测旋毛虫相关抗体的目的。结果　１９ 份人工感染鼠血清、５ 份人工感染猪血清、３ 份旋毛虫病猪血清、４ 份病人血清均呈阳性反应，而对照血清均为阴性反应；该方法在鼠感染旋毛虫后第五天可测出ＩｇＭ 抗体，第九天可测出ＩｇＧ抗体。结论　本检测技术简便易行，不需专用设备，可望成为一种快速诊断旋毛虫病的有效方法。Aim To establish a rapidly detecting method for the specific antibodies of Trichinella spiralis Method Latex was coated with p49/GST and polystyrene beads(Dynabeads)were coated with anti-antibodies SAM-IgG、SAH-IgG、RAP-IgG or SAM-IgM Then the test serum was incubated with two particles for 30 min at room temperature with slowly shaking After sedimentation of the polystyrene beads with a magnet,the turbidity of the resultant latex suspension was estimated with eyes or measured spectrophotometrically at a wavelength of 400nm Results Positive result of IgG antibody were found in 19 sera of experimental infected mice,3 sera of experimental infected swine and 4 sera of patients The IgM and IgG can be check up in mice after 5 and 9 days after experimental infection The result agreed with those obtained by ELISA Conclusion A two-particle turbidimetric latex immunoassy established by this study is a rapid,easy and precise method for the diagnosis of the trichinosis福建省科委(农医)项目!(95-Z150

DIAGNOSIS OF TRICHINOSIS BY ELISA WITH P49/GST ANTIGEN

目的以纯化的融合蛋白 p49/GST为抗原建立ELISA检测方法。方法对一批试验血清进行间接ELISA检测。结果 19份人工感染鼠血清、5份人工感染猪血清、4份旋毛虫病猪血清、4份病人血清呈IgG抗体阳性 ,2 1份人工感染鼠血清呈IgM抗体阳性 ,而正常对照血清及 30 0份屠宰场待检猪血清均呈阴性反应 ,其结果与常规压片法结果相符。结论融合蛋白p49/GST对于研制旋毛虫病的诊断抗原具有的潜在应用价值Aim To establish ELISA detecting method for the specific antibodies of Trichinella spiralis.Method A series of confirmed trichinosis sera were detected with ELISA with fusion protein p49/GST.Results Positive results of IgG antibody were found in 19sera of experimental infected mouse and 3sera of infected swine and 4 sera of patients;positive results of IgM antibody were found in 21 sera of experimental infected mouse .All normal sera were negative. Conclusion The ELISA with p49/GST can be regarded as a sensitive, specific immunological method for the diagnosis of trichinosis.福建省重点(农医 )项目资助!(项目号 :95 -Z -15 0

Application of biomarkers in marine environment monitoring

总结了国内外近十几年可应用于海洋环境污染监测常见的生物标记物及其主要的检测方法,对各自的应用特点进行了阐述,并展望了其应用的前景。其中对DNA损伤的检测作了较为详细的介绍。This paper sums up the recent-year application of biomarkers in marine environment motoring and the main detection methods of them.The features and the future prospects of these biomarkers are also discussed. The introduction of the DNA damage is the most detailed part in this paper.国家自然科学基金资助项目(A20077023,C40106012

LiFePO_4动力电池热物性测定及温升特性研究

温度是影响LiFePO4动力电池性能、安全及寿命的重要因素。电池热物性参数的测定及其温升特性的研究是电池热管理设计中的重要一环。通过实验测定了额定容量20 Ah的方块LiFePO4动力电池单体的比热容、生热速率、导热系数等重要热物性参数；并研究了强制风冷条件下，风速对电池在1、2、3电流倍率下的温升特性。电池热物性测定及温升研究将为电池散热设计提供重要的理论依据

Study on thermo-physical property measurement and temperature rise characteristic of LiFePO4 power battery

The temperature is an important factor affecting the performance, safety and cycle life of LiFePO4 power batteries. It is very important to measure the thermo-physical properties of batteries and study the temperature rise characteristic in the design o