Alternation of the Dissimilation Pathway for Glycerol and Amplification of Glycerol Dehydrogenase in Mutant Strains of Cellulomonas sp. NT3060

Mutant strain ME544, which is able to grow on glycerol slowly, was derived from glycerol-negative mutant strain G011, which is a derivative strain of Cellulomonas sp. NT3060 and is defective in both the enzyme activities of glycerol kinase and glycerol 3-phosphate dehydrogenase. The mutant strain still lacked both the enzyme activities involved in the dissimilation of glycerol and had the same level of glycerol dehydrogenase activity as the parent strain. Dihydroxyacetone kinase activity in mutant strain ME544 was inducibly formed, reaching 4-fold the level in mutant strain G011 in glycerol medium. Thus, the mutant strain seemed to dissimilate glycerol by means of glycerol dehydrogenase followed by an increase in dihydroxyacetone kinase. Subsequently, a mutant strain, GP1807, which was resistant to the inhibition of growth on glycerol by 1,2-propanediol, was derived from mutant strain ME544. Glycerol dehydrogenase activity of the mutant strain was amplified about 6-fold compared to that of the wild type strain.     

多杀性巴氏杆菌aroA基因缺失突变株的构建及鉴定

【目的】构建多杀性巴氏杆菌aroA基因缺失突变株,并验证其致病性。【方法】采用正向筛选同源重组技术构建多杀性巴氏杆菌aroA基因缺失突变株,利用PCR对突变株进行鉴定,分析其遗传稳定性、生长特性和致病性。【结果】成功构建多杀性巴氏杆菌aroA基因缺失突变株,连续传代20代,遗传稳定;突变株体外生长曲线表明,在前6h生长速度稍慢于亲本菌,随后两者生长速度一致。对小鼠的致病性试验表明:经腹腔注射aroA基因缺失突变株在1.0×106 CFU对小鼠无致死性,而亲本菌株在1.0×102 CFU对小鼠是致死性的。【结论】本研究获得多杀性巴氏杆菌aroA基因缺失突变株,对小鼠的致病性是减弱的。多杀性巴氏杆菌突变株的构建有助于研究其致病机理。     

以耐赖氨酸为标记的金霉素产生菌抗噬菌体变株的选育

本文以金霉素产生菌Ｆ－３０３作为出发菌株,通过ＵＶ＋ＥＭＳ复合处理,双层法不同梯度耐赖氨酸作标记,进行抗噬菌体的金霉素菌株选育,获得Ｆｓ－４８－１４菌株。该菌株不但表达了抗噬菌体的特性,而且产素效价超过Ｆ－３０３。通过无机盐正交条件试验,取得抗性变株最佳的发酵培养基配方。     

Infectivity and sensitivity to antibiotics of Rickettsia prowazekii mutants of the low-pathogenic strain E cultured in chick embryos

Selection of mutants of a low pathogenic strain E of R. prowazekii is a trend in genetic investigation of this Rickettsia species and one of the approaches to stabilizing the strain avirulent properties with a purpose of using in vaccine prophylaxis of typhus. The mutants of R. prowazekii, strain E selected by the authors earlier were characterized with respect to their infective capacity for chick embryos (CE) and antibiotic sensitivity. It was found that the infective capacity for CE of the erythromycin resistant mutant induced by nitroso guanidine (EErrI) was by ID50 2-3 logarithms lower than that of the initial strain E. The infective capacity for CE of the rifampicin resistant mutant induced by nitroso guanidine (ERifrI) and the spontaneous erythromycin resistant mutant was similar to that of strain E. The ERifrI strain differed from the initial strain E by higher sensitivity to tetracycline and erythromycin and the EErrI strain differed from the initial strain E by higher sensitivity to tetracycline and rifampicin. It was shown that the biological properties of the nitroso guanidine-induced mutants resistant to rifampicin and erythromycin differed from those of the initial strain E and the properties of the spontaneous erythromycin resistant mutant were similar to those of the initial strain E.     

Proline-hyperproducing strains of Serratia marcescens: enhancement of proline analog-mediated growth inhibition by increasing osmotic stress

Proline-producing strains of Serratia marcescens were more osmotolerant than wild-type strains. Growth inhibition by proline analogs was significantly enhanced by increasing the osmotic stress of the medium. Mutants resistant to azetidine-2-carboxylate were derived from a proline-producing strain, SP126, under a high osmotic condition. One of the mutants, strain SP187, produced 56 mg of L-proline per ml of medium containing sucrose and urea. This amount was ca. 3 times larger than that produced by strain SP126. The intracellular glutamate content which decreased in strain SP126 was restored in strain SP187. The glutamate dehydrogenase level of strain SP187 was 5 times higher than that of strain SP126.     

Influence of amino acids on the growth of Bacteroides melaninogenicus.

Addition of individual amino acids to a Trypticase-yeast extract-hemin medium affected growth rates and final yields of an asaccharolytic strain and a saccharolytic strain of Bacteroides melaninogenicus. L-Aspartate or L-asparagine produced maximal growth enhancement for both strains. L-[14C]aspartate was fermented by resting cells of the asaccharolytic strain. L-Cysteine or L-serine also enhanced growth for the saccharolytic strain. However, growth of the saccharolytic strain was inhibited by L-lysine, L-glutamate, L-glutamine, L-isoleucine, L-leucine, and L-proline; growth of the asaccharolytic strain was inhibited by DL-valine and L-serine. Both strains were inhibited by L-histidine, DL-methionine, L-tryptophan, L-arginine, and glycine.     

Demonstration of capsule in a strain of Staphylococcus hyicus by an electron microscope

A compact type variant designated as strain ST67V was isolated by a high-temperature subculture method from strain ST67P of Staphylococcus hyicus , which was a diffuse type in serum-soft agar. The parent strain was relatively virulent in mice and resisted ingestion by mouse peritoneal cells. The variant strain, however, was avirulent in mice and no antiphagocytic activity was observed in the peritoneal cavity. In ultra-thin sections of the organisms treated with anti-ST67P rabbit anti-serum conjugated with ferritin, the outermost layer of the cell wall of the parent strain was covered with a well-defined capsule while no capsule was shown in the variant strain.     

黑曲霉单宁酶高活性菌株的诱变选育*

以黑曲霉（Ａｓｐｅｒｇｉｌｌｕｓ ｎｈｉｇｅｒ）Ｎｏ．１３为出发菌株,经紫外线诱变处理,获得一株制备原生质体的起始菌,该菌株单宁酶活性比Ｎｏ．１３提高５５％,并对其制备原生质体的条件进行了研究,在优化方案基础上,紫外诱变原生质体,诱变株经筛选,最后得到一株具有稳定遗传性的单宁酶高活性菌株,在摇瓶培养基中进行生物转化实验,连续传代１０次,结果显示发酵液中没食子酸浓度始 维持在２２．８－２３．９ｍｇ／     

Proline-hyperproducing strains of Serratia marcescens: enhancement of proline analog-mediated growth inhibition by increasing osmotic stress.

Proline-producing strains of Serratia marcescens were more osmotolerant than wild-type strains. Growth inhibition by proline analogs was significantly enhanced by increasing the osmotic stress of the medium. Mutants resistant to azetidine-2-carboxylate were derived from a proline-producing strain, SP126, under a high osmotic condition. One of the mutants, strain SP187, produced 56 mg of L-proline per ml of medium containing sucrose and urea. This amount was ca. 3 times larger than that produced by strain SP126. The intracellular glutamate content which decreased in strain SP126 was restored in strain SP187. The glutamate dehydrogenase level of strain SP187 was 5 times higher than that of strain SP126.     

Separation of a phenol carboxylating organism from a two-member, strict anaerobic co-culture

In a culture converting phenol to benzoic acid under anaerobic conditions and previously described as being constituted of only a Clostridium-like strain 6, another bacterium (strain 7) was observed. Each organism was enriched by centrifugation on a Percoll gradient. Strain 6 was purified by dilution and plating. Strain 7 did not grow on solid media, but a strain 7 culture, cleared of strain 6, was obtained by subculturing in the presence of ampicillin and by dilution. In fresh medium, phenol was transformed by the reconstituted co-culture but not by each strain alone. In a supernatant from a co-culture or from a strain 6 culture, strain 7 alone transformed phenol but not strain 6. Maintenance of an active strain 7 in fresh medium instead of co-culture supernatant became possible when phenol was replaced by 4-hydroxybenzoate (4-OHB), which is decarboxylated to phenol before being transformed to benzoate. Even with 4-OHB, the use of co-culture (or strain 6 culture) supernatant resulted in faster transformation activity and growth rate. A phylogenetic analysis placed strain 7 in a cluster of uncultivated or nonisolated bacteria (92-96% homology). Strain 7 is also related to Desulfotomaculum, Desulfitobacterium, Desulfosporosinus, Moorella, and Sporotomaculum genera (87-92% homology).     

高效降解甲醛菌株的分离鉴定及其特性

首先对新分离的、能高效降解甲醛的两菌株A1和A2在形态学特征、生理生化特性及16S rDNA序列分析等方面进行了系统研究; 随后通过测定在液体培养过程中甲醛浓度的变化, 确定新分离菌株A1、A2降解溶液中甲醛的性能; 最后利用菌株A1、A2分别进行生物填料塔的挂膜实验, 确定其对甲醛气体的净化性能。结果表明: 菌株A1属于假单胞菌属(Pseudomonas), 菌株A2为鞘氨醇单胞菌属(Sphingomonas); 当甲醛初始浓度<1 200 mg/L时,菌株A1、A2都能完全降解溶液中的甲醛, 当甲醛浓度增高至1 600 mg/L时, 菌株A1在48 h后的甲醛降解率为50%, 菌株A2在104 h后的甲醛降解率为74.3%; 菌株A1、A2对甲醛气体的净化效率均能达到99%以上, 菌株A1的甲醛生化去除量能达到26.4 mg/(L?h), 菌株A2的甲醛生化去除量可达20.6 mg/(L?h)。     

Demonstration of capsule in a strain of Staphylococcus hyicus by an electron microscope

A compact type variant designated as strain ST67V was isolated by a high-temperature subculture method from strain ST67P of Staphylococcus hyicus, which was a diffuse type in serum-soft agar. The parent strain was relatively virulent in mice and resisted ingestion by mouse peritoneal cells. The variant strain, however, was avirulent in mice and no antiphagocytic activity was observed in the peritoneal cavity. In ultra-thin sections of the organisms treated with anti-ST67P rabbit anti-serum conjugated with ferritin, the outermost layer of the cell wall of the parent strain was covered with a well-defined capsule while no capsule was shown in the variant strain.     

产共轭亚油酸菌株的筛选及其等离子体诱变

本实验从东北酸菜汁中筛选获得了一株产共轭亚油酸能力较好的菌株SC-05,特别是气相色谱分析结果表明,合成的CLA产物构型为单一构型,仅为c9, t11-18：2-CLA,经鉴定该菌为乳杆菌属Lactobacillus sp.。菌株SC-05经低温等离子体处理后,筛选出突变株A-08,其共轭亚油酸产量达到119.24μg/mL,比出发菌株增加了83.0%,且突变株A-08具有较好的遗传稳定性。     

赤霉酸高产菌株的选育

本试验评估了赤霉酸生产菌株“４３０３”的生产性能,并以此菌株为出发菌株,进行了紫外线诱变育种,选出了一株赤霉酸高产菌株ＵＦ４５６。通过试验发现“４３０３”平均产量为１０３６±１２９ｍｇ／Ｌ,变异系数为０．１２,紫外线诱变对菌株产赤霉酸分布呈均匀状态。将“４３０３”与ＵＦ４５６进行赤霉酸产势比较,结果表明：ＵＦ４５６菌株平均产量较“４３０３”高２８．８％,产势高２０％。     

阿维菌素B1a组分高产菌株的定向选育

以阿维链霉菌(Streptomyces avermitilis)1-17为出发菌株,分别使用紫外线及亚硝基胍并结合L-异亮氨酸诱导手段进行诱变处理,得到AVMB1a组分摇瓶发酵水平较出发菌株提高12.86%的突变株3-6.传代实验表明该菌株的高产性能稳定.结果表明,采用UV、NTG诱变结合L-Ile诱导的手段可以获得B1a组分显著提高的菌株.     

模糊数学在平菇品种选育决策中的应用

应用模糊数学对 7个平菇品种进行模糊判决 ,结果表明 :在权重偏重于生物学效率的凸模糊判决中平菇 17- 2为首选。而在权重相等的模糊判决中平菇 99号为首选。对平菇 99号进行综合评判的结果为某类菇农对该品种很喜欢。     

复合诱变选育林肯霉素高产菌株

以林肯链霉菌9502(Streptomyces lincolnensis9502)为出发菌株,进行NTG诱变处理,并用高效的琼脂块培养法对菌株进行筛选,得到产林肯霉素相对效价提高35.4%的变异株9502-7。对9502-7菌株孢子采用紫外线处理,得到变异高产菌株9502-7-12,其相对效价较出发菌株提高50%以上。     

Comparative studies of strains Ictero No. I and RGA as the type strain of Leptospira interrogans: agglutinin absorption test, protein and antigen profiles, and enzyme activities

Strain Ictero No. I, the first isolate of Leptospira, isolated by Inada and Ido in 1914, was found to be sufficiently qualified to be the type strain of Leptospira interrogans rather than strain RGA. In an agglutinin absorption test, anti-Ictero No. I serum was not absorbed completely with strain RGA, and 25% of the homologous titer remained unabsorbed, while anti-RGA serum was completely absorbed with strain Ictero No. I. Thus, strain Ictero No. I was not serologically identical with strain RGA, and the two strains were considered to be different serovars. A protein band with a molecular weight of approximately 33,000 daltons was detected in strain Ictero No. I but not in strain RGA by SDS-PAGE. By Western blotting, this protein band was detectable with anti-Ictero No. I serum but not with anti-RGA serum. The presence of the 33K protein in strain Ictero No. I, but not in strain RGA, was confirmed by radioimmunoprecipitation using [125I]-labeled antigens, indicating that the protein antigen was surface-exposed. Only 8 of the 89 enzymes activities were different between strains Ictero No. I and RGA (line Sapporo). From the above results, we propose that strain Ictero No. I should be designated as the type strain of L. interrogans instead of strain RGA.     

蒜油对MRSA菌株的抗菌作用及作用机制研究

目的研究蒜油对MRSA菌株的抗菌作用及其作用机制,为治疗MRSA菌株感染提供实验依据。方法纸片法药敏试验测抑菌环直径,微量稀释法测其最低抑菌浓度(MIC),酶标仪测定不同时间MRSA菌株生长A640值,革兰染色镜检观察蒜油对MRSA菌株细胞壁的影响,SDS-PAGE凝胶电泳分析蒜油对MRSA菌株蛋白的影响。结果蒜油对MRSA菌株具有明显抗菌作用,其MIC和MBC分别为10mg/mL和20 mg/mL;1/2MIC和1MIC的蒜油作用MRSA菌株后,菌株生长曲线平缓,无对数期;且其对对数期细菌有明显杀菌作用;蒜油对MRSA菌株细胞壁无明显影响,凝胶电泳分析有特征条带出现。结论蒜油对MRSA菌株具有明显抗菌作用,其杀菌机制与蛋白合成有关。