两株高DNA拓扑异构酶Ⅰ抑制活性的药用植物内生真菌筛选

从7种中国南方药用植物中分离了278株内生真菌,分析了其发酵液甲醇提取物对人DNA拓扑异构酶Ⅰ(hTo-poI)松驰活性的抑制能力。对2株具有高hTopoI抑制力的内生真菌(LF4-7L和LF6-1)进行了深入研究。通过发酵物对hTo-poI抑制能力的时间变化曲线,表明2株内生真菌的hTopoI抑制物质在其生长停滞阶段才大量产生和积累。经分子鉴定,LF4-7L可能是阴炭团菌(Hypoxylon stygium),而与LF6-1L最相近的是毛竹基腐病菌(Arthrinium phaeospermum)。     

Isolation and Characterization of Bacterial Strains with Inulinase Activity

Thirty-two bacterial strains growing on inulin as the sole carbon and energy source were isolated from soil samples by enrichment culture on a mineral medium. Twenty of the strains were identified as Flavobacterium multivorum. All the bacteria contained a β-fructosidase that was active on both inulin and sucrose. The enzyme activity was cell bound and was produced at the end of the growth phase. These enzymes have potential uses in the preparation of fructose syrups from inulin and invert sugar from sucrose.     

Isolation and Characterization of Thermophilic Bacterial Strains with Inulinase Activity

Nine bacterial strains growing on inulin as the sole carbon and energy source were isolated from soil samples by enrichment culture on a mineral medium. Four of the strains were thermophilic and belong to the genus Bacillus. The thermophilic strains synthesized a β-fructosidase that was active on both inulin and sucrose. The presence of inulin in the culture medium is necessary for enzyme synthesis. Most of the activity on inulin was recovered in the culture medium, and the enzyme was synthesized during cell growth.     

具有免疫调节活性的果胶类物质

果胶类物质是广泛存在于植物细胞壁中的一类复杂多糖,对于植物生长与病害防御等起着重要的调节作用,随着糖类分离纯化和结构研究技术的进步,尤其随着糖类体内外活性测试方法的发展,果胶类物质的结构与活性研究得到了普遍重视。     

Biological Pretreatment with Two Bacterial Strains for Enzymatic Hydrolysis of Office Paper

The cellulose-hydrolyzing strains, Sphingomonas paucimobilis MK1 and Bacillus circulans MK2, were separated from soil and were grown together in a single culture plate. Growth B. circulans MK2 in liquid culture required symbiosis with S. paucimobilis MK1. Biological pretreatment with the combined strain suspension after the liquid culture improved enzymatic hydrolysis of office paper from municipal wastes. Sugar recovery by S. paucimobilis MK1 (51%) was 1.4 times higher than that of the untreated sample (30%) and in the strain combination with B. circulans MK2, recovery was further improved by 2.5 times (75%). The sugar recovery in maximum condition was enhanced up to 94% for office paper. Furthermore, biological pretreatment effects were confirmed for more than 1 day less time. In X-ray diffraction patterns for the crystallinity of cellulose in office paper changed after biological pretreatment, the crystallinity was increased in comparison to that in untreated paper. The mechanism of biological pretreatment effect was explained by the fact that the strain acted as an endoglucanase, which hydrolyzes amorphous areas randomly.     

两株具有抗癌活性内生细菌的分离及分类

从西双版纳植物样品爵床[Rostellularia procumbens(L.) Nees]中分离到2株具有较强抗癌活性的内生细菌YIM 56077和YIM 56081。通过对其进行表型特征、生长及生理生化特性、细胞化学组份以及系统进化分析,发现这2株菌与B.flexus IFO15715T的亲缘关系最近,但它们在生长、生理生化等特征上表现明显差异,是芽胞杆菌属的2个具有开发潜力的新菌株。     

Selection of Penicillium funiculosum Strains with High Glucose Oxidase Activity

Metabolic inhibitors, riboflavin, and end products of glucose oxidation were shown to be effective for the selection ofPenicillium funiculosum mutant strains with a high glucose oxidase activity. The incidence of positive mutations was highest in clones resistant to sodium azide, riboflavin, and -D-glucono--lactone. Enzyme activity in Penicillium funiculosum mutants was studied in submerged culture. The level of glucose oxidase synthesis in seven cultures was 24–56% higher than that in the parent strain of Penicillium funiculosum NMM95.132.     

2株海绵细菌的分离与鉴定

利用R2A培养基对海绵中的细菌进行分离,获得89株菌落形态有差异的菌株。通过在R2A培养基和营养丰富的LB培养基上的长势比较,发现有13株菌在LB培养基上生长缓慢、长势较弱。对此13株菌进行16S rDNA序列测定,发现菌株HB09009与Bacillus carboniphilus JCM9731T(AB021182)同源性最高,为97.1%;菌株HB09012与Planctomyces maris DSM8797T(NR025327)同源性最高,为97.4%,在发育树上处于一个分支,但在生长条件、培养特征和生理生化等方面都存在较大的差异,初步鉴定HB09009可能是Bacillus属的一个新种,暂定名为Bacillus sp.HB09009;鉴定HB09012可能是Planctomyces属的一个新种,暂定名Plancto-myces sp.HB09012。     

Reduction of Selenium Oxyanions in Wastewater Using Two Bacterial Strains

The biological reduction of selenium oxyanions is capable of reducing both selenate and selenite to insoluble elemental selenium. In this process, however, bacteria inevitably require expensive chemicals such as yeast extract in almost all cases. Therefore, the reduction of selenium oxyanions with inexpensive alcohol would be more practical. A Pseudomonas sp. strain 4C‐C isolated from a sludge in a wastewater treatment facility was able to reduce selenate to selenite using ethanol as an electron donor for its anaerobic respiration, but could not reduce selenite to elemental selenium. Paracoccus denitrificans JCM‐6892, on the other hand, was observed to be able to reduce selenite to elemental selenium in the presence of ethanol, but not selenate to selenite. Therefore, a mixture containing a suspension of Pseudomonas sp. strain 4C‐C and P. denitrificans JCM‐6892 cells allowed selenate to be reduced to insoluble elemental selenium via selenite in the presence of ethanol and was also capable of reducing nitrate to nitrogen gas. Aiming at simplicity of the recovery process of insoluble elemental selenium, a polymeric gel immobilized mixture of the two bacterial strains was examined using ethanol as an electron donor. The immobilized mixture could therefore reduce not only selenate to elemental selenium, but also nitrate to nitrogen gas in a single step. The gel that immobilized the microbial mixture changed its color during the process to bright red and no red elemental selenium was left in the wastewater. This indicates that the reduced elemental selenium was completely absorbed in the gel. This simple bacterial combination would therefore be effective in the presence of ethanol to reduce selenium oxyanions in various wastewaters containing selenium and the other oxyanions.     

Characterization of Two Paenibacillus amylolyticus Strain 27C64 Pectate Lyases with Activity on Highly Methylated Pectin

Two pectate lyases were identified from Paenibacillus amylolyticus 27C64; both enzymes demonstrated activity on methylated pectin in addition to polygalacturonic acid. PelA is in a subclass of the pectate lyase family III. PelB shows some features of pectate lyase family I but is highly divergent.Pectinases have many industrial applications, including uses in food and textile production (9, 12). Additionally, pectinases are important for the degradation of biomass, where pectin can comprise a significant portion of plant structure (5, 6). The degradation of pectin requires methylesterases and depolymerases. Pectin methylesterases are responsible for the hydrolysis of methylester linkages from the polygalacturonic acid (PGA) backbone (24), while pectin depolymerases act upon the polygalacturonate backbone and belong to one of two families, polygalacturonases or lyases. Polygalacturonases hydrolytically cleave the polygalacturonate chain, while lyases cleave by β-elimination, giving a Δ4,5-unsaturated product (10, 19). There are two types of lyases: pectate lyases (PLs), which cleave unesterified polygalacturonate, and pectin lyases, which cleave methylesterified pectin.Paenibacillus amylolyticus strain 27C64, isolated from the larval hindgut of the aquatic crane fly, Tipula abdominalis, possesses a wide range of lignocellulose-degrading enzymes. This study describes two pectate lyases from P. amylolyticus that display unusual activity by combining traits of pectate and pectin lyases (2, 7, 21, 22).     

好氧反硝化细菌N6-1亚硝酸盐还原活性的研究

针对本实验室筛选得到的1株好氧反硝化细菌(Aerobic Denitrifying Bacteria)N6-1,研究了培养条件和培养基组分对菌株亚硝酸盐还原活性的影响.结果表明,其最适培养条件为30℃、摇床转速120 r/min、初始pH 8.5,分别以NaNO2和乙酸钠为唯一氮源和碳源时,最适碳氮比为12;初始NaNO2浓度为2g/L时,培养20 h后NO2- -N全部被还原,平均还原速率可达20.3 mg/L·h;1.5%NaCl和1%蛋白胨对其亚硝酸盐还原活性没有明显影响;10 L发酵罐培养24 h后菌体浓度高达1.2×1011 CFU/mL.     

Aedesin: Structure and Antimicrobial Activity against Multidrug Resistant Bacterial Strains

Multidrug resistance, which is acquired by both Gram-positive and Gram-negative bacteria, causes infections that are associated with significant morbidity and mortality in many clinical settings around the world. Because of the rapidly increasing incidence of pathogens that have become resistant to all or nearly all available antibiotics, there is a need for a new generation of antimicrobials with a broad therapeutic range for specific applications against infections. Aedesin is a cecropin-like anti-microbial peptide that was recently isolated from dengue virus-infected salivary glands of the Aedes aegypti mosquito. In the present study, we have refined the analysis of its structural characteristics and have determined its antimicrobial effects against a large panel of multidrug resistant bacterial strains, directly isolated from infected patients. Based the results from nuclear magnetic resonance spectroscopy analysis, Aedesin has a helix-bend-helix structure typical for a member of the family of α-helix anti-microbial peptides. Aedesin efficiently killed Gram-negative bacterial strains that display the most worrisome resistance mechanisms encountered in the clinic, including resistance to carbapenems, aminoglycosides, cephalosporins, 4^th generation fluoroquinolones, folate inhibitors and monobactams. In contrast, Gram-positive strains were insensitive to the lytic effects of the peptide. The anti-bacterial activity of Aedesin was found to be salt-resistant, indicating that it is active under physiological conditions encountered in body fluids characterized by ionic salt concentrations. In conclusion, because of its strong lytic activity against multidrug resistant Gram-negative bacterial strains displaying all types of clinically relevant resistance mechanisms known today, Aedesin might be an interesting candidate for the development of alternative treatment for infections caused by these types of bacteria.     

青霉单宁酶高活性菌株的诱变选育

利用塔拉单宁诱导丝状真菌产生单宁酶的原理,通过富集培养,从天然源分离得到30株具有较高单宁酶活性的青霉菌;经二级发酵程序,对这30株菌进行了生物转化复筛实验,选择出能水解塔拉单宁,且生物催化活性较高的青霉野生株Penicilliumsp.No.23,对No.23进行经紫外诱变处理,诱变株经筛选,最后得到1株具有稳定遗传性的单宁酶高活性菌株,其单宁酶活性比出发菌株提高了35%。     

两株高丁醇耐受细菌的分离鉴定及其丁醇耐受特性的研究

高浓度丁醇耐受菌株是丁醇异源重组生产的关键因素.本研究对不同环境中耐受丁醇的微生物进行筛选,从自然环境中分离得到两株能够耐受高浓度丁醇的菌株,分别命名为btpz-4-1和btpz-6-3,它们耐受丁醇的浓度达到了25 g/L.通过分子标记物16S rDNA的鉴定以及分子系统进化树的分析,btpz-4-1被鉴定为Lactobacillus mucosae,btpz-6-3被鉴定为Pediococcus pentosaceus.同时,对它们的生理特性进行研究,结果显示btpz-4-1和btpz-6-3的最适生长温度分别为45℃和42℃,最适生长pH分别为6.0和6.5.     

Construction of Yeast Strains with High Cell Surface Lipase Activity by Using Novel Display Systems Based on the Flo1p Flocculation Functional Domain

We constructed a novel cell-surface display system, using as a new type of cell-wall anchor 3,297 or 4,341 bp of the 3′ region of the FLO1 gene (FS or FL gene, respectively), which encodes the flocculation functional domain of Flo1p. In this system, the N terminus of the target protein was fused to the FS or FL protein and the fusion proteins were expressed under the control of the inducible promoter UPR-ICL (5′ upstream region of the isocitrate lyase of Candida tropicalis). Using this new system, recombinant lipase with a pro sequence from Rhizopus oryzae (rProROL), which has its active site near the C terminus, was displayed on the cell surface. Cell-surface display of the FSProROL and FLProROL fusion proteins was confirmed by immunofluorescence microscopy and immunoblotting. Lipase activity reached 145 IU/liter (61.3 IU/g [dry cell weight]) on the surface of the yeast cells, which successfully catalyzed the methanolysis reaction. Using these whole-cell biocatalysts, methylesters synthesized from triglyceride and methanol reached 78.3% after 72 h of reaction. To our knowledge, this is the first example of cell-surface display of lipase with high activity. Interestingly, the yeast cells displaying the FLProROL protein showed strong flocculation, even though the glycosylphosphatidylinositol anchor attachment signal and cell-membrane-anchoring region of Flo1p had been deleted from this gene. The cell-surface display system based on FL thus endows the yeast strain with both novel enzyme display and strong flocculation ability.     

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／     

细菌好氧反硝化研究进展

阐述了好氧反硝化细菌的种类及有关性质 ,并从电子理论、氧的浓度、反硝化酶系等方面对细菌好氧反硝化的作用机制进行了探讨 ,对细菌好氧反硝化的研究概况、进展及其研究意义和目前存在的问题作了较为详细的介绍。     

Polygalacturonase, Pectate Lyase and Pectin Methylesterase Activity in Pathogenic Strains of Phytophthora capsici Incubated under Different Conditions

Pectolytic enzymes are found mainly in fungi and bacteria. The most widely occurring enzymes are polygalacturonase (PGs), pectin methylesterase (PMEs) and pectate lyase (PLs) produced during the infection process and during culturing. The secretion of these enzymes results in the disorganization of the plant cell walls, which is responsible for the pathogenicity of the pathogens. These enzymes degrade the pectin of plants causing maceration of plant tissues and the enzyme activity increases under favourable environmental conditions. We have found that Phytophthora capsici , a pathogenic oomycete, produces levels of these three enzymes equal to those produced by soft-rotting Erwinia chrysanthemi . The activity of PGs, PLs and PMEs was investigated at the optimum temperature, pH and ionic strength in highly pathogenic P. capsici strains cultivated in two kinds of liquid medium containing either crude pepper extracts plus pectin or pectin as the carbon source. Virulence tests and enzymes activity showed that there was a high correlation between the enzyme activity and the pathogenicity of P. capsici . The effects of different carbon sources on the enzyme activity showed that pepper extract plus pectin was the best source for the carbon source.