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Applied Biochemistry and Microbiology, Vol Oleg Mosin Julia
Applied Biochemistry and Microbiology, Vol. 35, ffo. /.
1999, pp. 29-17. Translated from Prikladnayti Biokhimiya i Mikrobialogiya, Vol. 35, No. 1,@ 1999, pp. 34-42. Original Russian Copyright
© /999 hy Mosin, Skluclnev, Shvatz.
Incorporation of [2,3,4,5,6- 2
H 5 ]Phenylalanine,
[3,5- 2
H 2 ]Tyrosine, and [2,4,5,6,7 2 H 5
]Tryptophan
into the Bacteriorhodopsin Molecule of
Halobacterium halobium
O. V. Mosin*, D. A. Skladnev**, and V. I. Shvets** Lotnonosov Moscow State Academy of Fine Chemical Technology, Moscow, 117571 Russia
**
State Center of Genetics and Selection of Industrial Microorganisms (GNU GENETICA), Moscow, 113515 Russia
Received September 25, 1997
Abstract —
Incorporation of [2,3A5,6- 2 H 5
]phenylalanine, [3,5- 2 H
2 ]tyrosine, and [2,4,5,6,7- 2
H 5 ]tryptophan
into the bacteriorhodopsin molecule followed by semipreparative isolation of bacteriorhodopsin resulted in a yield of 8-10 mg per g bacterial biomass.
This method is based on the growth of the strain of halophilic bacteria Halobacterium halobium on a synthetic medium containing
2 H-labeled aromatic ammo acids and fractionation
of solubilized (in 0.5% sodium dodecyl sulfate) protein by methanol, including purification of carotenoids. lip-
ids, and high-molecular-weight and low-molecular-weight compounds, as well as gel-permeation chromatog- raphy on Sephadex G-200. Incorporation of
2 H-labeled amino acids was analyzed by electron impact mass
spectrometry after hydrolysis of the protein in 4 N Ba(OH) 2 and separation in the form of methyl esters of
/V-DNS derivatives



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