Chapter VIII Isolation, Growth and Requirements of Halophilic Bacteria

Gibbons, N. E.

doi:10.1016/s0580-9517(08)70507-5

Cited by 28 publications

(20 citation statements)

References 24 publications

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“…In the author's laboratory, both halobacteria and halococci have been routinely grown in a SO-gal stainless steel fermentor (Stainless & Steel Products Co., St. Paul, Minnesota) under aseptic conditions at 37°C, using 160 liters of medium 1 and 2-S% inoculum. The cell yield was about the same as that reported by Gibbons (1969). The halo bacteria reached maximum growth after 3 days, the halococci after S-6 days.…”

Section: Medium 5 For Growth Of Halobacterium Volcaniisupporting

confidence: 85%

“…Methods for the isolation, cultivation and maintenance of the Halobacteriaceae have been reviewed by Eimhjellen (1965) and by Gibbons (1969). These reviews are somewhat more detailed than the following.…”

Section: Isolation and Cultivationmentioning

confidence: 99%

“…Discussing medium 7, Gibbons (1969) points out that KOH used in adjusting pH adds a significant amount of potassium, which otherwise limits the growth. Adding 2-S g/liter KCI still improves growth.…”

Section: Medium 5 For Growth Of Halobacterium Volcaniimentioning

confidence: 99%

“…The air flow was 0.1 (vol/vol) per min, CHAPTER 78 and foaming was prevented by adding Dow-Coming Antifoam A. Halobacterium cutirubrum was reported to reach maximum growth after 3-S days at 37°C; the cell yield was 1.0-1.S g dry weight/liter. Gibbons (1969) used 100 liters of medium 2 in a ISO-liter stainless steel fermentor and S% inoculum. He reported a cell yield of S-6 g wet weightlliter after 6S h at 3rc.…”

Section: Medium 5 For Growth Of Halobacterium Volcaniimentioning

confidence: 99%

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The Family Halobacteriaceae

Larsen¹

1981

The Prokaryotes

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Section: Medium 5 For Growth Of Halobacterium Volcaniisupporting

confidence: 85%

Section: Isolation and Cultivationmentioning

confidence: 99%

Section: Medium 5 For Growth Of Halobacterium Volcaniimentioning

confidence: 99%

Section: Medium 5 For Growth Of Halobacterium Volcaniimentioning

confidence: 99%

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The Family Halobacteriaceae

Larsen¹

1981

The Prokaryotes

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“…These bacteria were found to require high salt concentrations, and later Gibbons noted that the extremely halophilic bacteria could only survive in solutions containing more than 15% (wt/vol) NaCl (3). He suggested that this requirement for salt and the production of red pigment could be used as taxonomic characteristics to classify certain bacteria as extreme halophiles (3).…”

mentioning

confidence: 99%

Halomonas elongata, a New Genus and Species of Extremely Salt-Tolerant Bacteria

Vreeland

Litchfield

Martin

et al. 1980

International Journal of Systematic Bacteriology

389

199

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Membrane‐bound proteinases of Halobacterium halobium

Fricke

Parchmann

Aurich

1993

J. Basic Microbiol.

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Cell envelopes of Halobacterium halobium contain proteolytic activities, which cleave '*'I-insulin and 14C-casein with pH-optima in the alkaline range. About 50% of the intracellular proteases are strongly attached to the cell envelopes and cannot be removed by washing with 25% sodium chloride solution. This high amount of membrane-bound proteases is independent of the kind of cultivation. The main part of the envelope-bound proteolytic activity is attributed to metalloproteases, with EDTA and o-phenanthroline causing inhibition. Serine and cysteine protease inhibitors had no influence.In density gradients (consisting of Ficoll and 20% sodium chloride) proteolytic activities could be detected in two distinct bands, a minor band at a density of 1.165 and the main proteolytic activity at 1.145 g/cm3. When the cell wall was removed from the other cell envelope constituents by EDTA treatment, a single band with proteolytic activity in the density gradient at 1.145 g/cm3 was detectable.Lubrol, Brij 35 , and octyl glucoside were used for the solubilization of the membrane-bound proteases. The greatest part of them could be solubilized using 0.2% octyl glucoside.14C-labelled bacteriorhodopsin was not cleaved by any of the intracellular or extracellular proteinases.Halophilic archaebacteria have a well fitted metabolism for growth on proteins, peptides and amino acids. Most of the strains tested secrete caseinolytic and gelatinolytic proteases into the medium (GIBBONS 1969, GUTIERREZ and GONZALEZ 1972). Up to now only a few proteases of this kind have been isolated and characterized. Most proteins of these bacteria, including the proteases, are strongly halophilic and irreversibly denaturated at low salt concentrations (HOLMES et al. 1965, LEICHT et al. 1978, WERBER et al. 1986, NORBERG and HOFSTEN 1969. Purification steps must therefore be performed in the presence of high salt concentrations. NORBERG and HOFSTEN (1970) used hydroxylapatite chromatography for the efficient purification of an extracellular protease of Halobacterium salinarium. An extracellular serine protease of H . halobium cleaving casein and L-pyro-glutamyl-Ala-AlaLeu-pNA was also isolated by bacitracin-Sepharose affinity chromatography (IZOTOVA et al. 1983). Recently a halophilic serine protease of a Halobacterium strain was purified with HPLC-methods (SCHMITT et al. 1990) using azocoll as substrate.The known extracellular proteases are caseinolytic or able to cleave synthetic peptide substrates (IZOTOVA et al. 1983, NORBERG andHOFSTEN (1969). Up to the present only extracellular serine proteases have been purified and characterized. In addition, NORBERG and HOFSTEN (1969) described a particle-bound protease, differing in its inhibition pattern from the extracellular serine protease. They found that 90% of the intracellular activity was bound to the cell envelope. Further characterization and solubilization of this enzyme were not possible, because the sensitivity of the azocasein assay used was too low.Bacteriorhodopsin is the main protein of the pu...

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Chapter VIII Isolation, Growth and Requirements of Halophilic Bacteria

Cited by 28 publications

References 24 publications

The Family Halobacteriaceae

The Family Halobacteriaceae

Halomonas elongata, a New Genus and Species of Extremely Salt-Tolerant Bacteria

Membrane‐bound proteinases of Halobacterium halobium

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