枯草杆菌BS-98分泌的抗真菌蛋白的分离纯化及其部分性质的研究

拮抗菌（Bacillussubtilis）BS-98菌株在BPY液体培养基中产生的蛋白质经硫酸铵分级盐析、SephsdexG-100柱层析、DEAE-纤维素柱层析和SephadexG－100柱再层析后,分离纯化得到的抗菌蛋白在电泳中显现出三条带。经初步纯化的抗菌蛋白对热稳定,对蛋白酶部分敏感。抑菌谱测定表明,抗菌蛋白对芦笋茎枯病菌、小麦赤霉病菌、棉花枯萎病菌、棉花黄萎病菌、灰霉菌、立枯丝核菌等病原真菌及黄瓜角斑病菌都具有强烈的抑制作用。     

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02抗菌蛋白的分离纯化和其性质的研究

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02,是一株能强烈抑制玉米纹枯菌（Rhizoctonia cerealis）、白色念珠菌 (Candidal vaginitis)等病害真菌的拮抗菌株。SS02菌株培养液经硫酸铵分级盐析,Sephadex G-75凝胶柱层析和DEAE-32纤维素柱层析分离纯化出一种抗菌蛋白,命名为SD22。蛋白电泳分析结果表明,此蛋白分子量为56000,等电点为6.4。此蛋白对热和紫外线稳定,对蛋白酶部分敏感。纯化后的SD22对玉米纹枯菌(Rhizoctonia cerealis)、油菜菌核病(Sclerotinia sclerotiorum)、苹果轮纹病菌（Physalospora piricala）、绿色木霉(Trichodema viride)、绿色粘帚霉（Gliocladium viride）、弯孢霉菌(Curvularia leaf spot)、镰刀霉菌(Fusarium sp.)、赤霉菌(Fusarium head blight)、球孢白僵菌（Beauveria bassiana）、大肠杆菌（Escherichia coli）、金黄色葡萄球菌（Staphylo-coccus aureus）、枯草芽孢杆菌（Bacillus subtilis）、白色念珠菌(Candidal vaginitis）、冬瓜枯萎菌（Fusarium oxysporum Schl.emend.Snyder & Hanse）等菌有很强的抑制作用。     

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02抗菌蛋白的分离纯化和其性质的研究

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02,是一株能强烈抑制玉米纹枯菌（Rhizoctonia cerealis）、白色念珠菌 (Candidal vaginitis)等病害真菌的拮抗菌株。SS02菌株培养液经硫酸铵分级盐析,Sephadex G-75凝胶柱层析和DEAE-32纤维素柱层析分离纯化出一种抗菌蛋白,命名为SD22。蛋白电泳分析结果表明,此蛋白分子量为56000,等电点为6.4。此蛋白对热和紫外线稳定,对蛋白酶部分敏感。纯化后的SD22对玉米纹枯菌(Rhizoctonia cerealis)、油菜菌核病(Sclerotinia sclerotiorum)、苹果轮纹病菌（Physalospora piricala）、绿色木霉(Trichodema viride)、绿色粘帚霉（Gliocladium viride）、弯孢霉菌(Curvularia leaf spot)、镰刀霉菌(Fusarium sp.)、赤霉菌(Fusarium head blight)、球孢白僵菌（Beauveria bassiana）、大肠杆菌（Escherichia coli）、金黄色葡萄球菌（Staphylo-coccus aureus）、枯草芽孢杆菌（Bacillus subtilis）、白色念珠菌(Candidal vaginitis）、冬瓜枯萎菌（Fusarium oxysporum Schl.emend.Snyder & Hanse）等菌有很强的抑制作用。     

内芽孢杆菌属细菌(Paenibacillus daejeonensis)SS02抗菌蛋白的分离纯化和其性质的研究

内芽孢杆菌属细菌（Paenibacillus daejeoneusis）SS02,是一株能强烈抑制玉米纹枯菌（Rhizoctonia cerealis）、白色念珠菌（Candidal vaginitis）等病害真菌的拮抗菌株。SS02菌株培养液经硫酸铵分级盐析,SephadexG-75凝胶柱层析和DEAE-32纤维素柱层析分离纯化出一种抗菌蛋白,命名为SD22。蛋白电泳分析结果表明,此蛋白分子量为56000,等电点为6．4。此蛋白对热和紫外线稳定,对蛋白酶部分敏感。纯化后的SD22对玉米纹枯菌（Rhizoctonia cerealis）、油菜菌核病（Sclerotinia sclerotiorum）、苹果轮纹病菌（Physalospora piricala）、绿色木霉（Triclwdema viride）、绿色粘帚霉（Gliocladium viride）、弯孢霉菌（Curvula rialeafspot）、镰刀霉菌（Fusarium sp．）、赤霉菌（Fusarium headblight）、球孢白僵菌（Beauveria bassiana）、大肠杆菌（Escherichia coli）、金黄色葡萄球菌（Staphylo-coccus aureus）、枯草芽孢杆菌（Bacillus subtilis）、白色念珠菌（Candidal vaginitis）、冬瓜枯萎菌（Fusarium oxysporum Schl．emend,Snyder＆Hause）等菌有很强的抑制作用。     

芒果炭疽病菌β-微管蛋白基因的克隆及其与多菌灵抗药性发生的关系

参照豆科合萌属（Aeschynomene）作物炭疽病菌的tub1和tub2基因序列设计了2对引物,分别从芒果（Mangifera）炭疽病菌对多菌灵(MBC)田间抗药性（MBCR）和敏感（MBCS）的菌株中扩增β_微管蛋白基因。结果只有以tub2为参照设计的引物扩增到了特异片段。进一步对全基因进行了克隆和测序。该基因序列全长1344bp,编码447aa,其核苷酸和氨基酸序列与豆科合萌属炭疽病菌的tub2基因高度同源。对芒果炭疽病菌抗、感菌株β_微管蛋白氨基酸序列进行比较分析,发现第181、237和363位氨基酸发生了突变,而其它位置（如第198位或200位）均不变。     

真菌还原Cr(Ⅵ)的研究

从不同来源的样品中分离筛选出几株抗Cr（Ⅵ）的真菌,他们能在含300—500mg／LK2Cr2O7的蔗糖合成培养基中生长,其中BS－1菌株抗K2Cr2O7达900mg／L．BS－1等4株真菌在含200mg／LK2Cr2O7的培养基中生长4-6d后,培养液中的Cr（Ⅵ）已全部消失。这些真菌经鉴定为青霉菌（Penicilliumsp．）BS－1和BS－3,黑曲霉（Aspergillusniger）BR-4和黄曲霉（Aspergillusflavus）BX－1。经紫外可见光扫描及化学分析证实,高毒的Cr（Ⅵ）可被真菌还原成为低毒的Cr（Ⅲ）。BS－1菌株细胞还原Cr（Ⅵ）的最适温度为30℃,最适pH7．0。葡萄糖（0．25％）对细胞还原Cr（Ⅵ）有促进作用,但高浓度的Cr（Ⅵ）则抑制细胞对Cr（Ⅵ）的还原。     

抗菌蛋白LCIIB的纯化及性质

枯草杆菌A014培养液经硫酸铵沉淀、CM—52纤维素柱层析、FPLG的Mono S和Superose12柱层析,分离纯化出一种抗菌蛋白,命名为LCIIB。电泳分析结果表明该蛋白的分子量为22500Da,等电点为9,95。LCIIB含19种不同氨基酸,缺少半胱氨酸。自动Edman降解法从N端测出24个氨基酸残基,经计算机分析,表明与2万多种已知蛋白没有有效同源性。纯化后的LCIIB对水稻白叶枯病菌有很强的抑制作用,当浓度在5／μmol／L时,能抑制50％此菌G株系的生长。     

链霉菌S01菌株几丁质酶对植物病原真菌的拮抗作用

纯化后的链霉菌S01菌株几丁质酶用环柱法在PDA平板上对杨树腐烂病菌（Valsasordida）、葡萄孢菌（Botrytissp．AS3．266）、黄瓜黑腥病菌（Cladosporiumcucumerinrm）、辣椒疫病菌（Phytophlhoracopsici）、棉花黄萎病菌（Verliilliumalbo-atrum)、立枯丝核菌（Rhizoctonasolani）等植物病原真菌及产黄青霉（Penlcillumc     

肉色杯伞抗炎蛋白的提纯及某些理化性质

通过硫酸铵沉淀和DEAE一Sephadcx A-25柱层析等步骤,从肉色杯伞(Clitocybe geotropa子实体中,获得抗炎蛋白质晶体,纯化后的抗炎蛋白质为针状结晶,平均收率为40 mg／0.5kg．经聚丙烯酰胺盘状电泳鉴定为单一区带,Sephadex G-100柱层析UV 280 nm吸收峰为均一峰形。分子量3万左右。据测定,其N-末端氨基酸为缬氨酸。该蛋白由17种氨基酸按不同比例构成,其中苏氨酸居首位,其次为丝氨酸。此肽不含糖。     

地衣芽孢杆菌β－甘露聚糖酶的纯化及酶学性质

地衣芽孢杆菌（Ｂａｃｉｌｌｕｓｌｉｃｈｅｎｉｆｏｒｍｉｓ）ＮＫ－２７菌株发酵产生的β－甘露聚糖酶（β－ｍａｎｎａｎａｓｅ）经硫酸铵盐析沉淀,两次ＤＥＡＥ纤维素和ＳｅｐｈａｄｅｘＧ－１００离子交换柱层析以及制备ＰＡＧＥ筹步骤,获得了凝胶电泳均一的样品。用ＳＤＳ－凝胶电泳测得纯化后的β－甘露聚糖酶分子量为２６ｋＤ,用凝胶聚焦电泳测得等电点ＰＩ为５．０。酶反应的最适ｐＨ为９．０,最后温度为６０℃,稳定ｐＨ为６．０—９．０,稳定温度为４０℃。金     

Cicerarin,a novel antifungal peptide from the green chickpea

A peptide designated cicerarin, with an N-terminal amino acid sequence VKSTGRADDDLAVKTKYLPP dissimilar from known proteins and peptides and a molecular mass of 8kDa, was isolated from seeds of the green chickpea Cicer arietinum cv green chickpea. Cicerarin was isolated with a procedure that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, and gel filtration by fast protein liquid chromatography on Superdex 75. Cicerarin was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel in 10mM Tris-HCl buffer (pH 7.3). Cicerarin exerted antifungal activity against Botrytis cinerea, Mycosphaerella arachidicola, and Physalospora piricola. The antifungal activity was preserved after exposure to 100 degrees C for 15min.     

三种苔藓植物提取物对植物病原菌的抑菌性研究

以立枯丝核菌(Rhizoctoniasolani)、棉花枯萎病菌(Fusariumoxysporum)、苹果轮纹病菌(Physalosporapiricolanosa)、草莓灰霉病菌(Botrytiscinerea)四种植物病原菌为供试病原菌,对大镰刀藓Drepanocladusexannulatus、锐尖匍灯藓Plagiomniumacutum和疣小金发藓Pogonatumurnigerum三种藓类醇提液进行抑菌活性筛选,结果表明,大镰刀藓提取液对立枯丝核菌有较好的抑制作用,对立枯丝核菌的EC50为0.878mg/mL;而锐尖匍灯藓的提取液对立枯丝核菌的生长却有促进作用。     

Purification of allivin,a novel antifungal protein from bulbs of the round-cloved garlic

A novel antifungal protein, designated allivin, was isolated from bulbs of the round-cloved garlic Allium sativum var. round clove with a procedure involving ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-Sepharose and FPLC-gel filtration on Superdex 75. Allivin possessed an N-terminal sequence demonstrating very little similarity to sequences of Allium sativum chitinases and ribosome inactivating proteins. Allivin exhibited a molecular weight of 13 kDa in gel filtration and SDS-polyacrylamide gel electrophoresis. It displayed antifungal activity against Botrytis cinerea, Mycosphaerella arachidicola and Physalospora piricola. It inhibited translation in a cell-free rabbit reticulocyte system with an IC50 of 1.6 microM.     

Alveolarin, a novel antifungal polypeptide from the wild mushroom Polyporus alveolaris

An antifungal polypeptide, with a molecular mass of 28 kDa as judged by gel filtration and appearing as a single band with a molecular mass of 14 kDa in sodium dodecyl suflate-polyacrylamide gel electrophoresis, was isolated from fresh fruiting bodies of the mushroom Polyporus alveolaris. The antifungal polypeptide, designated as alveolarin, demonstrated an inhibitory action on mycelial growth in Botrytis cinerea, Fusarium oxysporum, Mycosphaerella arachidicola and Physalospora piricola. Alveolarin was isolated with a procedure that entailed ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, and gel filtration on Superdex 75 by fast protein liquid chromatography.     

Isolation of a novel thermolabile heterodimeric ribonuclease with antifungal and antiproliferative activities from roots of the sanchi ginseng Panax notoginseng.

An isolation procedure, consisting of ion exchange chromatography on CM-Sepharose, affinity chromatography on Affi-gel blue gel, and fast protein liquid chromatography on Mono S, was utilized to purify a base-nonspecific, heterodimeric ribonuclease (RNase) with diverse activities from roots of the sanchi ginseng Panax notoginseng. The RNase is unique in that it consists of two different nonglycoprotein subunits with a molecular weight of 27 and 29 kDa, respectively. The latter subunit is characterized by an N-terminal sequence showing remarkable similarity to that of the bitter gourd RNase. The Panax notoginseng RNase demonstrates potent RNase and translation-inhibitory activities. In addition, it exhibits antiproliferative activity toward leukemia L1210 cells and antifungal activity against Physalospora piricola and Coprinus comatus. Its RNase activity is not heat-resistant, unlike most RNases which are thermostable.