Bacillus stearothermophilus

Biological indicators for steam sterilization utilize bacillus stearothermophilus. In monitoring industrial cycles, a sufficient number of preparations each having a known degree of resistance are added to the load and retrieved after exposure, and cultured. 

An alternative approach to the microbial deracemization of secondary alcohols is to use two different microorganisms with complementary stereoselectivity. Fantin et al. studied the stereoinversion of several secondary alcohols using the culture supernatants of two microorganisms, namely Bacillus stearothermophilus and Yarrowia lipolytica (Figure 5.18) [31]. The authors tested three main systems for deracemization. First, they used the supernatant from cultures of B. stearothermophilus, to which they added Y. lipolytica cells and the racemic alcohols. Secondly, they used the culture supernatant of Y. lipolytica and added B. stearothermophilus cells and the racemic alcohols. Finally, they resuspended the cells of both organisms in phosphate buffer and added the racemic alcohols. The best results were obtained in the first system with 6-penten-2-ol (26) (100% ee and 100% yield). The phosphate buffer system gave... 

Wang et al. report the death kinetics of Bacillus stearothermophilus spores using wet, thermal sterilization. Twenty minutes at 110°C reduces the viable count by a factor of lO". The activation temperature, E/Rg, is 34,200 K. How long will it take to deactivate by a factor of lO ... 

FIG. 1 Freeze-etching image of a bacterial cell of (a) Desulfotomaculum nigrificans (bar, 100 nm). Atomic force micrographs of the S-layer proteins of (b) Bacillus sphaericus CCM 2177 and (c) Bacillus stearothermophilus PV72/p2 recrystallized in monolayers on silicon wafers. Bars, 50 nm. The insets in (b) and (c) show the corresponding computer-image reconstructions. 

FIG. 3 Three-dimensional model of the protein mass distribution of the S-layer of Bacillus stearothermophilus NRS 2004/3a [(a) outer, (b) inner face]. The square S-layer is about 8 nm thick and exhibits a center-to-center spacing of the morphological units of 13.5 nm. The protein meshwork composed of a single protein species shows one square-shaped, two elongated, and four small pores per morphological unit. (Modified from Ref. 7.)... 

Biological indicators (Bis) for use in thermal, chemical or radiation sterilization processes consist of standardized bacterial spore preparations which are usually in the form either of suspensions in water or culture medium or of spores dried on paper, aluminium or plastic carriers. As with chentical indicators, they are usually placed in dummy packs located at strategic sites in the sterilizer. Alternatively, for gaseous sterihzation these may also be placed within a tubular hehx (Line-Pickerill) device. After the sterilization process, the aqueous suspensions or spores on carriers are aseptically transferred to an appropriate nutrient medium which is then incubated and periodically examined for signs of growth. Spores of Bacillus stearothermophilus in sealed ampoules of cultrrre medium are used for steam sterilization morritoring, and these may be incubated directly at 55°C this eliminates the need for an aseptic transfer. 

Low temperature steam (73°C) and formaldehyde (12mg - ) Bacillus stearothermophilus 5min... 

Hoxey E.V., Soper C.J. Davies D.J.G. (1984) The effect of temperature and formaldehyde concentration on the inactivation of Bacillus stearothermophilus spores by LTSF. J Pharm... 

Zaar A, W Eisenreich, A Bacher, G Fuchs (2001) A novel pathway of aerobic benzoate catabolism in the bacteria AzoarcMS evansii and Bacillus stearothermophilus. J Biol Chem 276 24997-25004. 

The preceding protocol can be successfully applied, essentially without modifications, to prepare active cell-free extracts from bacteria other than Escherichia coli (e.g., Bacillus stearothermophilus and clinical isolates of Pseudomonas aeruginosa bearing multiple antibiotic resistance). 

This essential property of IF2 can be tested in at least three different ways, all of which require the availability of f[3H]Met-tRNA and IF2, which are prepared according to the protocol detailed in Milon et al. (2007). However, all the tests described in this section can make use of the sturdier and smaller C domain of Bacillus stearothermophilus IF2, since this domain contains all molecular determinants for the IF2-fMet-tRNA interaction (Guenneugues et al, 2000 Spurio et al, 2000). The method for the preparation and purification of B. stearothermophilus IF2C is essentially that described by Spurio et al. (1993). The concentration of the protein... 

Packman, L.C., and Perham, R.N. (1982) Quaternary structure of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus studied by a new reversible cross-linking procedure with bis(imidoesters). Biochemistry 21, 5171-5175. 

Kennedy LD (1974) Teichoic-acid synthesis in Bacillus-stearothermophilus. Biochem J... 

Figure 2.45 shows a typical set of D-values (time in which the number of bacteria is reduced by one decade) for Bacillus Stearothermophilus for different surface temperatures of the parts to be sterilized and the necessary concentrations of H202. 

Bacillus stearothermophilus, 11 35, 48 thermal resistance characteristics, 8 634t... 

Some recent approaches to orientated immobilization exploited methods of genetic engineering to introduce suitable binding site like a cysteine residue [101]. Mansfeld and Ulbrich-Hofmann introduced cysteine on a thermolysinelike neutral protease from Bacillus stearothermophilus [114]. This approach is especially interesting since it allows the introduction of cystein residues in different positions of the protein molecule and therefore enables one to study the effects of molecule orientation on its biological activity [115]. More information about the oriented immobilization can be found in recent reviews of Turkova [101,110]. 

The cell walls of most Gram-positive bacteria contain (1 ->3) poly(glycerol phosphate) teichoic acids which can bear D-alanyl or glycosyl substituents.56 However, the cell walls of Bacillus stearothermophilus B65 contain a glycerol teichoic acid which is free of substituents.57 This is contrary to evidence that had been obtained from biosynthetic studies.58... 

Inorganic pyrophosphatase Bacillus stearothermophilus 7.9 90 Morita and Mathemeier... 

The only other E. coli ribosomal protein whose crystallization has so far been reported is L29 (Appelt et al., 1981). On the other hand, attempts to crystallize ribosomal proteins from the thermophilic Bacillus stearothermophilus have been more successful. Protein BL17, which according to its amino acid sequence (Kimura et al., 1980) corresponds to protein L9 from the E. coli ribosome (Kimura et al., 1982), was the first intact ribosomal protein to give crystals useful for X-ray structural analysis (Appelt et al., 1979). Several other B. stearothermophUus ribosomal proteins, namely BL6 and BL30 (Appelt eteU., 1981,1983) from the large and BS5 (Appelt et al., 1983) from the small subunit have been crystallized, and the determination of their three-dimensional structure at a resolution of better than 3 A is now in progress. Furthermore, crystals of aB. stearothermophilus ribosomal protein complex, which corresponds to the complex (L7/L12)4 LIO from E. coli ribosome, have been obtained (Liljas and Newcomer, 1981). 

Fig. 7. (a) Crystals of E. colt 70 S ribosomes, (b) and (c) Electron micrographs of sections through three-dimensional crystals shown in (a) in two orthogonal directions (Wittmann et al., 1982). (d) and (e) Crystals and computed filtered image of a section through a crystal of Bacillus stearothermophilus 50 S ribosomal subunits (Yonath et al., 1982a,b Leonard et al., 1983). (d) and (e) are related to two different crystal forms. Reproduced with permission from Wittmann (1983). 

Figure 4 A library of Mj TyrRS mutants was generated by randomizing five residues in the tyrosine binding site. The crystai structure of Bacillus stearothermophilus TyrRS is shown with residues labeled in biack. Corresponding residues in the Methanococcus Jannaschii TyrRS are labeled in blue. Reproduced from L. Wang A. Brock B. Herberich P. G. Schultz, Science 2001, 292, 498-500, with permission from AAAS.

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