Bacillus subtilis expression

Wu, X.C., Lee, W., Tran, L. and Wong, S.L. (1991) Engineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases. Journal of Bacteriology, 173 (16), 4952 1958. 

Wong, S. L., Ye, R., andNathoo, S. (1994). Engineering and production of streptokinase in a Bacillus subtilis expression-secretion system. Appl. Environ. Microbiol., 60(2), 517-523. 

Lauraeus M, Haltia T, Saraste M, Wikstrom M (1991) Bacillus subtilis expresses two kinds of haem A-containing terminal oxidases. Eur J Biochem 197 695-705... 

The Gram-positive bacteria Bacillus subtilis, Bacillus megaterium and Bacillus brevis are also used for heterologous expression. Terpe [13] presented a table with some pharmaceutically and industrially relevant proteins expressed with this host. It is the most used prokaryotic host after... 

Wu, S.C. and Wong, S.L. (1999) Development of improved pUB 110-based vectors for expression and secretion studies in Bacillus subtilis. Journal of Biotechnology, 72 (1-2), 185-195. 

Nijland, R., Veening, J.W. and Kuipers, O.P. (2007) A derepression system based on the Bacillus subtilis sporulation pathway offers dynamic control of heterologous gene expression. Applied and Environmental Microbiology, 73 (7), 2390-2393. 

An (5)-specific alcohol dehydrogenase gene from Rhodococcus erythropolis and GDH from Bacillus subtilis were ligated into one plasmid, which was expressed in Escherichia coli strain DSM14 459 to provide an (S)-selective whole-cell catalyst. 

Several methods for performing the test have been described, the guideline describes the diffusion and suspension test. Bacteria should be exposed to the test substance both in the presence and absence of an appropriate metabolic activation system. The response is expressed in the preferential inhibition of growth or the preferential killing of the DNA repair deficient strain. Escherichia coli polA (W3110/p3478) or Bacillus subtilis rec (H17/M45) pairs are recommended. The test should initially be performed over a broad range of concentrations. 

FIGURE 6.14 (a, c) Raman spectra of dried bacterial spores Bacillus subtilis), and (b, d) the corresponding polarized CARS spectra of the same bacterial spores in aqueous solution. (From Petrov, G. I., Yakovlev, V. V., Sokolov, A. V., and Scully, M. O., Optics Express, 13(23) 9537-9542, 2005. Used with permission.)... 

Yamamoto, H. Murata, M. Sekiguchi, J. The CitST two-component system regulates the expression of the Mg-citrate transporter in Bacillus subtilis. Mol. Microbiol., 37, 898-912 (2000)... 

N. The essential two-component regulatory system encoded by yycF and yycG modulates expression of the ftsAZ operon in Bacillus subtilis. Microbiology, 146, 1573-1583 (2000)... 

The host bacteria used for production of recombinant proteins are usually E. coli, or Bacillus subtilis they may express proteins at 1 % to over 50% of the cellular protein, depending on such variables as the source, promoter structure, and vector type. Generally the proteins are expressed intracellularly, but leader sequences for excretion may be included. In the latter case, the protein is generally excreted into the periplasmic space, which limits the amount that can be produced. Excretion from grampositive species such as B. subtilis sends the product into the culture medium, with little feedback limitation on total expression level. 

Apply a standard error-prone PCR (epPCR see Chapter 2) to the wild-type lipase gene from Bacillus subtilis and express conventionally in E. coli [37] initiate by inoculation of the cultures in deep-well microtiter plates (96-well format). Use LB/M9 medium with 100 pL carbenicillin (lOOmgmL-1) per 100 mL of medium and incubate for 5-6 h at 37 °C while shaking. 

Application and Principle This procedure is used to determine the a-amylase activity, expressed as bacterial amylase units (BAU), of enzyme preparations derived from Bacillus subtilis var., Bacillus licheniformis var., and Bacillus stearoth-ermophilus. It is not applicable to products that contain 13-amylase. The assay is based on the time required to obtain a standard degree of hydrolysis of a starch solution at 30° 0.1°. The degree of hydrolysis is determined by comparing the iodine color of the hydrolysate with that of a standard. 

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