提高Lovastatin产生菌生物合成能力的研究

用微波等离子体(N^ 20w，4win)诱变M．ruber，筛选到一株高产菌株，再经过发酵工艺优化，最终Lovastaitin的发酵产率提高21．8％。     

Plasmid transformation in Bacillus subtilis NB22, an antifungal-antibiotic iturin producer

A transformation system with plasmids was developed for Bacillus subtilis NB22, an antibiotic iturin producing strain. Treatment of B. subtilis NB22 with 4 M KCl was effective for the induction of competence, followed by uptake of plasmid DNA in the presence of polyethylene glycol. The efficiency of transformation of this bacterium with pC194 and pUB110 was 4.1 X 10(3) and 1.5 X 10(3) transformants per micrograms DNA, respectively and the transformation frequency was 3.3 X 10(-3) and 7.2 X 10(-4), transformants per viable cell, respectively. This method was much faster and three orders of magnitude more efficient in transformation efficiency than protoplast transformation methods.     

Engineering modular ester fermentative pathways in Escherichia coli

Sensation profiles are observed all around us and are made up of many different molecules, such as esters. These profiles can be mimicked in everyday items for their uses in foods, beverages, cosmetics, perfumes, solvents, and biofuels. Here, we developed a systematic ‘natural’ way to derive these products via fermentative biosynthesis. Each ester fermentative pathway was designed as an exchangeable ester production module for generating two precursors− alcohols and acyl-CoAs that were condensed by an alcohol acyltransferase to produce a combinatorial library of unique esters. As a proof-of-principle, we coupled these ester modules with an engineered, modular, Escherichia coli chassis in a plug-and-play fashion to create microbial cell factories for enhanced anaerobic production of a butyrate ester library. We demonstrated tight coupling between the modular chassis and ester modules for enhanced product biosynthesis, an engineered phenotype useful for directed metabolic pathway evolution. Compared to the wildtype, the engineered cell factories yielded up to 48 fold increase in butyrate ester production from glucose.     

Pathogenic potential and horizontal gene transfer in ovine gastrointestinal Escherichia coli

Aims: To demonstrate that a thorough characterization and virulotyping of Escherichia coli strains isolated from sheep over time leads to new insights into ovine E. coli potentially becoming human pathogens through horizontal gene transfer. Methods and Results: One hundred and fifty E. coli isolates from two sheep, sampled over 3 weeks, were characterized by serotyping, virulotyping, genotyping using multiple locus variable number tandem repeats analysis (MLVA) and susceptibility to phage infection in vitro. The 35 MLVA profiles and the serotype and virulotypes of the strains were closely associated. Many MLVA profiles differed in one locus independent of serotypes. Escherichia coli isolates of the same serotype or virulotype had identical or very similar MLVA profiles. No transductants that incorporated the bacteriophages were found in vivo, but six E. coli isolates were susceptible to the phage infection in vitro. Changes in MLVA profiles were seen after acquisition of Stx phages in vitro only. Conclusions: The sheep carried Stx phage susceptible E. coli that possessed virulence markers associated with human pathogenicity. Changes in bacterial genomes by phage transfer may complicate outbreak source investigations. Serotype has to be taken into account when evaluating strain relationships by MLVA. Significance and Impact of the Study: Sheep carry E. coli that encode for virulence markers and belong to serogroups known to be human pathogens. In addition, a selection of isolates was found to be susceptible to horizontal transfer of Shiga toxin genes by means of bacteriophages in vitro, and the transfer resulted in a discernible change of the MLVA patterns of E. coli.     

产木聚糖酶白地霉培养特性及部分纯化的酶学特性

本文对白地霉Ref1的培养特性、产酶条件和酶学特性进行了初步研究。结果表明:该菌为低温型菌株,其最佳生长条件为pH6、20℃和酵母膏作为氮源;最佳产酶条件为pH3-7、15℃及以酵母膏氮源;条件优化后产酶可达118.7U/mL,可溶蛋白含量可达到60μg/mL,酶溶液的比活可达到1250U/mg蛋白质;该木聚糖酶的最适反应温度和pH分别为50℃和5,金属离子Mg2+、Na+和8mmol/L的Fe2+、Cu2+、Zn2+等对木聚糖酶的活性有抑制作用,而Ca2+、4mmol/L的Fe2+、Cu2+、Zn2+和8mmol/L的Mn2+等对该酶反应则有促进作用;该木聚糖酶在保温2h后在15-40℃范围内能保持80%以上的酶活性,在50℃时能保持68%的酶活性;用lineweaver-Burk作图法(双倒数作图法)求得该酶的最大反应速度Vmax和Km值分别为163.38mmol/mg/min和0.75mg/mL。     

中国稻谷可培养真菌群落组成及多样性

真菌生长导致粮食品质下降及真菌毒素污染,是造成粮食损失的主要原因。我们采用形态学与分子系统发育分析相结合的方法,对中国13个省78份稻谷样品的真菌群落组成及多样性进行研究。结果表明,本次共分离622株真菌,经初步鉴定分属于17属73个物种。南北方因气候条件差异较大,稻谷真菌数量和物种组成都存在很大差异。南方稻谷表面真菌数量（10 ⁴cfu/g）明显高于北方（10 ³cfu/g）。北方收获期的优势种主要属于枝孢属Cladosporium,南方收获期的优势种为黄曲霉Aspergillus flavus、黑曲霉Aspergillus niger和阿姆斯特丹曲霉Aspergillus amstelodami等。进入储藏期后,由于中国仓储管理要求较高,储藏环境相对稳定（aw≤0.75）,耐旱真菌阿姆斯特丹曲霉和多育曲霉Aspergillus proliferans（占比40%以上）成为优势种,这类真菌可作为储藏期粮堆异常的早期预警指示菌。南方收获期感染的黄曲霉和黑曲霉进入储藏期后仍保持优势比例,导致储藏期间毒素污染风险必然增加,因此南方稻谷收储环节均应加强监管。本研究通过分析南北方收获和储藏环节稻谷真菌群落组成差异及产毒真菌的分布规律,以期为中国粮库在稻谷收储环节的真菌毒素防控提供依据。