Breeding of a non-urea producing sake yeast with killer character using a kar1-1 mutant as a killer donor

Arginase-deficient (car1/car1) sake yeasts can brew sake without urea, a main precursor of ethyl carbamate, which is a suspected carcinogen in various fermented beverages. For the use of car1/car1 yeasts in sake production, contamination by wild-type (CAR1/CAR1) yeasts is a major problem. To protect sake mash against such contamination, killer character was introduced into the car1/car1 sake yeast HL163 by rare mating and protoplast fusion, using a kar1–1 haploid harboring killer dsRNA plasmids as a killer donor. All killer yeasts obtained showed no arginase activity and the same DNA content per cell as strain HL163, and produced sake with ordinary quality and very low levels of urea. We also demonstrated that one of these killer yeasts could effectively eliminate contaminant cells of a CAR1/CAR1 yeast from sake mash. Journal of Industrial Microbiology & Biotechnology (2000) 24, 203–209. Received 09 August 1999/ Accepted in revised form 07 December 1999     

日本亮耳菌——一种新的杀线虫真菌

从贵州省宽阔水自然保护区采到的日本亮耳菌Lampteromycesjaponicus(Kawam.)Sing.这一中国新记录种,其菌丝体及其丙酮粗提物对线虫有毒杀作用,是一类有应用前景的食线虫真菌资源。     

氨基酸产生菌E_(65)中质粒的分离与鉴定

在棒杆菌与短杆菌中,用微量质粒分离方法筛选到一株含多种质粒的乙酰短杆菌(Brevibacterium actylicum)E_(65)菌株,该菌株具有产氨基酸特性。进一步大量制备其质粒DNA,并进行氧化绝密度梯度超离心纯化,以及电泳分析,电镜观察,并对质粒进行分离与鉴定,从而确定了每种质粒的分子量大小。     

Molecular cloning of the delta-endotoxin gene of Bacillus thuringiensis var. israelensis

A transformant of Bacillus megaterium, VB131, was isolated which carries a 6.3-kb XbaI segment of the crystal toxin gene of Bacillus thuringiensis var. israelensis (BTI) cloned in a vector plasmid pBC16 to yield pVB131. The chimeric plasmid DNA from VB131 was introduced into a transformable Bacillus subtilis strain by competence transformation. Both the B. megaterium VB131 strain and the B. subtilis strain harboring the chimeric plasmid produced irregular, parasporal, phase-refractile, crystalline inclusions (Cry+) during sporulation. The sporulated cells as well as the isolated crystal inclusions of the pVB131-containing B. megaterium and B. subtilis strains were highly toxic to the larvae of Aedes aegypti. Also, the solubilized crystal protein preparation from VB131[pVB131] showed clear immuno cross-reaction with antiserum to the BTI crystal toxin. 32P-labeled pVB131 plasmid DNA showed specific hybridization with a 112-kb plasmid DNA of Cry+ strains of BTI, and no hybridization with other plasmid or chromosomal DNA of either Cry+ or Cry- variants. These results are in agreement with our previous findings (González and Carlton, 1984) that the 112-kb plasmid of BTI is associated with the production of the crystal toxin.     

Ultrasensitive detection of Shiga toxin 2 and its variants in Shiga toxin‐producing Escherichia coli strains by a time‐resolved fluorescence immunoassay

A rapid and sensitive two‐step time‐resolved fluorescence immunoassay (TRFIA) was developed for the detection of Shiga toxin 2 (Stx2) and its variants in Shiga toxin‐producing Escherichia coli (STEC) strains. In sandwich mode, a monoclonal antibody against Stx2 was coated on a microtiter plate as a capture antibody. A tracer antibody against Stx2 labeled with europium(III) (Eu³⁺) chelate was then used as a detector, followed by fluorescence measurements using time‐resolved fluorescence. The sensitivity of Stx2 detection was 0.038 ng/ml (dynamic range, 0.1–1000 ng/ml). The intra‐ and inter‐assay coefficients of variation of the assay were 3.2% and 3.6%, respectively. The performance of the established assay was evaluated using culture supernatants of STEC strains, and the results were compared to those of a common HRP (horseradish peroxidase) labeling immunosorbent assay. A polymerase chain reaction (PCR) for the detection of genes encoding Stx1 and Stx2 was used as the reference for comparison. Correlation between the Stx2‐specific TRFIA and PCR was calculated by the use of kappa statics, exhibiting a perfect level of agreement. The availability of the sensitive and reliable Stx2‐specific TRFIA method for quantifying Stx2 and its variants in STEC strains will complement bacteria isolation‐based platform and aid in the accurate and prompt diagnosis of STEC infections.