生物化学实验课教学改革及本科生基本科研素质培养

生物化学实验课是生物学及相关专业的一门学科类通修课程,其教学目的是让学生学习和掌握生物化学实验基本操作技能的同时,注重学生基本科研素质培养。我们根据这个理念将生物化学实验课内容设计成三个模块:基本生物化学实验技术训练模块,综合实验训练模块,以及自主设计实验模块,并在课程开展中注重学生基本科研素质的培养,达到了实验教学培养学生科研实践能力和综合素质的目的

Optimization of solid culture conditions for TNF-α inhibitor qiaonanmycin via response surface methodology

目的利用响应面法对潜在的Tnf-α抑制剂桥南霉素固态发酵培养基进行优化。方法用HPlC做检测方法,以桥南霉素的洗脱峰面积为指标,先通过PlACkETT-burMAn(P-b)实验筛选出影响固态发酵产量的主要因素,进而进行单因素优化实验和响应面中心组合设计实验,确定主要影响因素的最佳浓度,得到最优的固体发酵培养基组成。结果优化后的发酵培养基组成:土豆400g/l、葡萄糖35g/l、蛋白胨10.5g/l、(nH4)2SO4 0.6g/l。按此配方固态发酵10l,对得到的粗提物进行HPlC检测,桥南霉素的实际产量为4.3Mg/l,与模型的理论值基本相符,较优化前的产量1.5Mg/l提高了186.7%。结论响应面方法是一种有效的培养基优化方法,经此优化方法桥南霉素的产量得到了较大的提高。Objective The solid fermentation culture conditions of compound qiaonanmycin were optimized via response surface methodology.Methods Based on quantitative detection method of qiaonanmycin by HPLC, the key fermentation factors were determined via P-B test, then their optimized concentrations were confirmed through response face method.Results The optimum solid fermentation culture medium compositions of qiaonanmycin were: potato 400g/L, dextrose 35g/L, peptone 10.5g/L, and(NH4)2SO4 0.6g/L.Under this condition, the yield of qiaonanmycin was 4.3mg/L, resulting in a 186.7% increase comparing with original production yield, and the optimized yield was consistent well with theoretical value predicted by response face method.Conclusion The optimized solid fermentation culture conditions were reliable and repeatable, it was warranted for large-scale fermentation process.中央高校基本业务费(No.2010121092); 厦门市科技计划项目(No.3502Z20123010

Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter

Agence Nationale de Recherche (ANR); Centre National de la Recherche Scientifique (CNRS); University of StrasbourgBackground: Death-Associated Protein Kinase (DAPK) is a member of the Ca(2+)/calmodulin regulated serine/threonine protein kinases. Its biological function has been associated with induced cell death, and in vivo use of selective small molecule inhibitors of DAPK catalytic activity has demonstrated that it is a potential therapeutic target for treatment of brain injuries and neurodegenerative diseases. Methodology/Principal Findings: In the in vitro study presented here, we describe the homodimerization of DAPK catalytic domain and the crucial role played by its basic loop structure that is part of the molecular fingerprint of death protein kinases. Nanoelectrospray ionization mass spectrometry of DAPK catalytic domain and a basic loop mutant DAPK protein performed under a variety of conditions was used to detect the monomer-dimer interchange. A chemical biological approach was used to find a fluorescent probe that allowed us to follow the oligomerization state of the protein in solution. Conclusions/Significance: The use of this combined biophysical and chemical biology approach facilitated the elucidation of a monomer-dimer equilibrium in which the basic loop plays a key role, as well as an apparent allosteric conformational change reported by the fluorescent probe that is independent of the basic loop structure

Four New Citrinin Derivatives from a Marine-Derived Penicillium sp Fungal Strain

863 project [2006AA09Z410, 2007AA091503]; Ministry of Science and Technology; Fundamental Research Funds for the Central Universities [2011121037]Four new citrinin derivatives, including two citrinin dimers and two citrinin monomer derivatives, were isolated and identified from a marine-derived fungal strain Penicillium sp. ML226 along with six known related compounds. Their structures were elucidated by spectroscopic and chemical methods. The new compounds showed modest cytotoxic activity against HepG-2 cell line and weak antimicrobial activity against Staphylococcus aureus

Study on the Isolation Identification and Antitumor Activity of a Phenol Derivative from Mangrove Fungus BYY-1

[作者简介] 杜希萍( 1978—) ，女，讲师，从事天然产物化学方面的研究． 通讯作者: 苏文金( 1956—) ，男，教授，从事海洋药物与食品方向研究，E-mail: wjsu@ jmu. edu. cn．[中文文摘]采用硅胶柱层析、薄层层析、Sephadex LH-20柱层析从红树内生真菌BYY-1的次级代谢产物中分离到化合物B1.通过乙酰化、NMR、HR ESI-Q-TOF MS等技术鉴定了化合物B1的结构,确定其为首次从红树内生真菌中分离到的化合物.运用MTT法和流式细胞术研究了化合物B1的抗肿瘤活性,结果表明,化合物B1可显著抑制Hela细胞的增殖[英文文摘]Compound B1 was obtained from the secondary metabolites of mangrove fungus BYY －1 using silica gel column chromatography，thin layer chromatography and Sephadex LH － 20 column chromatography．The structure of compound B1 was elucidated by acetylation，NMR and HR ESI －Q －TOF MS analysis，and compound B1 was isolated from mangrove fungi for the first time． The antitumor activity of compound B1 was studied by the methods of MTT and flow cytometric DNA analysis． The results showed compound B1 could significantly inhibit the proliferation of Hela cells with IC50 3. 3 μg /mL，and induce evident Hela cells apoptosis when compound B1 was 4 μg /mL．中央高校基本业务费资助项目(2010121092); 福建省科技计划重点项目(2011N1008);; 集美大学博士启动基金资助项目(ZQ2011004

5-(Hydroxymethyl)furan-2-carboxylic acid

Fundamental Research Funds for the Central Universities, China [2010121092]In the title compound, C(6)H(6)O(4), the furan ring is nearly coplanar with the carboxyl group, the maximum atomic deviation being 0.0248 (9) A. The crystal packing is stabilized by classical O-H...O and weak C-H...O hydrogen bonding

Diversity and bioactivities of marine actinobacteria from marine sediments of Fujian coast

目的探索福建沿海沉积物中可培养海洋放线菌的多样性及其抗菌、抗肿瘤活性,为发现新的药物先导化合物提供新菌源。方法从15份福建沿海沉积物选择性分离放; 线菌;16S; rRNA基因序列分析多样性;采用10种活性筛选模型分析生物活性。结果共分离到216株海洋放线菌,MOPS-脯氨酸培养基的分离效果最好。选取77株; 代表菌株进行16S; rRNA基因序列分析,发现其归属于14个属,其中10株为潜在新种。有164株(77%)海洋放线菌对8种指示菌中的一种或几种具有抑制作用;有39株; (18.1%)对HeLa细胞或HepG2细胞有细胞毒活性(抑制率≥50%)。结论福建沿海沉积物样品中含有丰富多样的海洋放线菌,大量菌株具有抗菌与; 抗肿瘤活性,是天然产物开发的重要资源。Objective To find new lead compounds, the diversity, antimicrobial and; antitumor activities of marine actinobacteria from marine sediments of; Fujian coast were investigated. Methods Actinobacteria strains were; selective isolated from 15 marine sediments of Fujian coast. The 16S; rRNA gene sequences analysis of representative strains were carried out; to evaluate their diversity. All of the isolates were assessed using 10; screening models for their bioactivities. Results A total of 216 marine; actinobacteria were isolated by using four media. The isolation effect; of MOPS-proline medium was the best choice. Phylogenetic analysis based; on 16S rRNA gene sequences showed that 77 representative strains; belonged to 14 genera, and there were 10 strains identified as potential; novel species. One hundred and sixty-four (77%) strains showed; antimicrobial activity against at least one kind of microorganisms.; Thirty-nine (18.1%) strains showed cytotoxic activity with inhibition; rate above 50% against Hela or HepG2 cells in vitro. Conclusion This; study indicated that actinobacteria from marine sediments of Fujian; coast are abundant, many of them had antimicrobial and cytotoxic; activities, which can be further explored.厦门市科技计划项

Replica Plating and Gene Screening Combined to Separate AHBA Synthase Gene Positive Actinomycetes

通信作者：[email protected]基因筛选是筛选具有抗生素产生潜力微生物的新方法.针对筛选中单菌落分离纯化不仅工作量大,而且耗费时间,采用一种将平板影印技术与PCR扩增技术相结合从土壤中快速获得具有安莎类抗生素产生潜力的放线菌的方法.根据安莎类抗生素合成途径中的3-氨基-5-羟基苯甲酸合成酶(AHBAs)基因的保守性,通过放线菌的培养、影印、AH-BAs基因的PCR扩增,已从33份土样中获得8株AHBAs阳性菌株.结果表明:该方法是一种非常有效的能够快速获得AHBAs基因阳性菌株的方法,并且该方法可扩展用于从土壤中分离其他有价值的抗生素产生菌.[英文文摘]Gene mining is a new strategy to discover microbial natural product,However,it is laborious to inculate and purify tested strains before gene amplification,so gene mining can not be applied to large amounts of samples.Herein,a high throughput method for the gene mining of soil actinomyetes, combining the replica plating techique and PCR 　amplification was recommended.In the present work,3-amino-5-hydroxybenxoic acid synthase(AHBAs) were used as amplicated gene.Through soil actinomyctes culture,colony replica planting,PCR amplification of AHBAs genes,isolation of positive strains and characterization of fermentation extracts, eight AHBAs positve strains were selected from 33plants rhizosphere soil samples.The results showed that the replica plating-gene screening combined technique was useful for the rapid selection of ansamycin-producing actinomycetes,and it could be applied to search for producers of other antibiotics in natural soils.中央高校基本业务费资助项目(2010121092,2011121037);科技部国际科技合作项目(2007DFA30970

Antitumor Activity of Mangrove Endophytic Fungi

作者简介: 郑忠辉(1956-),男, 教授[中文文摘]从红树植物秋茄、桐花树和白骨壤树皮样品中分离出125株内生真菌。采用MTT法对其体外抗肿瘤活性进行检测,发现11株红树植物内生真菌对KB(人口腔上皮癌)或HL-60(人白血病)细胞具有显著的抑制活性,占总分离菌株的8.8％。秋茄、桐花树和白骨壤的抗肿瘤活性菌株分别占供测菌株的11.6％、5.7％和4.8％。抗肿瘤活性菌株主要分布在拟青霉属及曲霉属。在所分离的菌株中,归属于拟青霉属的活性菌株占供测菌株的比例最高、活性最强。结果显示红树植物内生真菌是潜在的抗肿瘤药物资源。[英文文摘]One hundred and twenty-five strains of endophytic fungi were isolated from the inner barks of three kinds of mangrove plants,Kandelin canda,Avicennia marina and Aegiceras corniculatum.Antitumor activity was studied by the MTT assay.8.8% of endophytic fungi fermentation broths displayed cytotoxic activity on KB or HL-60 cells at and below a dilution of 1:100,11.6% cytotoxic-positive strains isolated from Kandelin canda,5.7% from Avicennia marina,and 4.8% from Aegiceras corniculatum.The active strains obtained mostly belong to Paecilomyces and Aspergillllus.Among all endophytic fungi isolated ,the genus Paecilomyces has the highest cytotoxic-positive rate and antitvmor activity.These results indicate mangrove endophytic fungi could be a promising source for antitumor bioactive agents.国家“863”计划(2001AA620401);福建省自然科学基金(C0210001

The Effects of Different Media on Antitumor Active Substances Production of Mangrove Fungus HTF3

对红树林真菌HTF3菌株在PD、GPY、YES、GPM及SYE 5种不同培养基中的生长情况、抗肿瘤活性及代谢产物的形成进行了研究,旨在了解培养基对该菌株生长及抗肿瘤活性物质合成的影响.结果显示:在不同的培养基中,HTF3菌株的生长速率不同,抗肿瘤活性及抗瘤谱也有明显差异;采用HPLC对不同培养基发酵液乙酸乙酯抽提物进行分析,发现不同抽提物的色谱峰数、色谱峰的保留时间及紫外吸收值均有较大的不同.实验结果表明:不同培养基对HTF3菌株抗肿瘤活性物质的合成有显著影响.Mangroves are special woody plant communities in the intertidal zone of tropical and subtropical coasts.Mangrove fungi growing in mangroves are attractive because a variety of chemically interesting and biologically significally substances have been purified from their secondary metabolites.Therefore,this paper studied the growth,antitumor activities and metabolites production of mangrove fungus HTF3 on potato dextrose medium,glucose peptone yeast extract medium,yeast extract sucrose medium,glucose peptone malt extract medium and starch yeast extract medium,to find out the effects of different media on the growth and antitumor active substances production of HTF3.The results showed that the growth rates of HTF3 on different solid media were different;the growth rates of HTF3 on glucose peptone yeast extract medium and starch yeast extract medium was 17.1mm/d and 11.1 mm/d;antitumor activities and antitumor specffum were markedly different in different liquid media,the IC_(50) against Raji cell of five extracts from different fermentation broth,namely A(PD),B(GPY),C(YES),D(GPM) and E(SYE) is 31.9,66.5,76,49.6 and 27.6 μg/mL respectively.The ethyl acetate extracts of the fermentation broth from different media were analysed using HPLC.The numbers of chromatographic peak,retention times and UV spectra were significant different.The study indicated diffetent media greatly affected the antitumor active substances production of HTF3.国家863计划(2001AA620401);; 福建省科技计划项目(2004N003)资