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[575.113:579.873.6]:552.578.3(282.256.

341)
alk- Rhodococcus erythropolis

.., .., .., ..

,
,
88% ().
. ,
,
[1].
n- ()
-.
,
O2 .

NADH (nicotinamide adenine dinucleotide) FAD
(flavin adenine dinucleotide) ,
: R-CH3 + O2 + NAH(P)H + H+ R-CH2OH + NAD(P)+ +
H2O.
[2].

Pseudomonas putida [3]. ,
(alk- ,
),

[3].

, P. putida.
, alk-
[2, 3],
Rhodococcus.
,
, ,
[4].
alk- -
.

2008-2009 .
. ..

8.

8 1 100 (
, ),
GTTP 0,2 (Millipore).

(Raymond, 1961) (/): KH2PO4 2,0;
CaCl22H2O 0,01; MgSO47H2O 0,2; FeSO4 0,01; NH4NO3 2,0;
MnSO45H2O 0,02; Na2HPO4 3,0; Na2CO3 0,1; 1,5; pH = 7,2.
168
,

(-
).

.
.
1 5 1,5
Eppendorf 4 .
50% -70 .


- [5].
(30 ): 94 , 2 ;
92 , 45 ; 52 , 45 ; 72 , 60 ;
72 , 60 ; 72 , 2 (1 ).

27f,

1350r,

16S [6, 7].


1,5%-
. ,
895-1118 ..
NCBI
BLASTN ClustalW v.1.4,
GA v.4.0
neighbor-joining.

PINTAIL (http://www.cardiff.ac.uk/biosi/research/biosoft).
alk-
-, , [8]:
AlkB-1

Alk-1F 5-CATAATAAAGGGCATCACCGT-3
Alk-1R 5-GATTTCATCCTCGAAACTCCAAAC-3
AlkB-2

Alk-2F 5-GAGACAAATCGTCTAAAACGTAA-3

Alk-2R 5-TTGTTATTATTCCAACTATGCTC-3
AlkB-3
Alk-3F 5TCGAGCACATCCGCGGCCACCA-3
Alk-3R 5-CCGTAGTGCTCGACGTAGTT-3

NCBI : 16S HQ702468,
HQ702469, HQ702470, HQ702471, HQ702472; alk- HQ702473,
HQ702474, HQ702475, HQ702477.
alk-. 4
alk- (Illumina GAIIx, ).
10 ..

Velvet_1.1.02 3897 1,8 . ..


6,9 .
Expasy Translate tool
(http://web.expasy.org/translate).

R.

erythropolis

PR4

NCBI.

BLASTX.
ClustalW v.1.4.

GA v.4.0 neighborjoining.

, , 5
alkB-,
. , 2

alkB1- alkB3-.

alkB3-.

alkB-
Rhodococcus. 16S

( ), alk- 4 5
(. 1). , ,
alk-
Rhodococcus.

alk-


3, .
4 5, (. 2),
,
, 3
.
25-6 3. ,

Rhodococcus, .
alkB- 4. AlkB-

Pseudomonas

putida

GPo1):

n-.

H138E[L/M]XHK143,

E167HXXGHH173 L309QRH[S/A]DHHA317. ,
(Identification of an amino acid..., 2005),

, , . ,

, , Fe(II) [9, 10].


L309QRH[S/A]DHHA317
alk-
, Rhodococcus
opacus B4. , ,
,

, ,
: (7 913 450 ..) (GC 67,9%),
pROB01 (558 192 ..) pROB02 (244 997 ..)
pKNR (111 160 ..), pKNR01 (4 367 ..) pKNR02 (2
773 ..). R. erythrypolis PR4,
1 [11],
(6 516 310 ..) (GC 62,31%),
(pREL1: 271 577 ..) (pREC1: 104 014 ..
pREC2: 3 637 ..). ,
. , alk-

, . alk-

[12].
,

n-,

alkB- [13]. , R.
erythropolis NRRL B-16531 Q15 4 alkB- [12].
,

,
Rhodococcus.

alkB- R. erythropolis 4
R. erythropolis PR4
NCBI,
6,5 [11]. 4
-1- ,

-1-

NCBI ( , . 19): 1
alk-1-monoox (BAH33111), 2 alk-1-monoox (BAH31254), 3 alk-1-monoox
(BAY32870), 4 alk-1-monoox (BAH36166).
alkB-, , 4 , ,
.
.

NCBI,

, R. erythropolis 4

HELGHK,

EHNHGHH

LQRHSDHHA. EHNFGHH -1 (BAH36166)


() (F).
R. erythropolis SK121 (04385381) R.
erythropolis PR4 (002768905), 94-97%.
, alk- 4
-1-, n, ,
(3,5oC) (90 .).

NCBI
-1- R. erythropolis 4

Rhodococcus.

, ,
, ,
,
. .. , alk-

. ,
Geobacillus Rhodococcus [12].
4

R.

erythropolis

SK121

PR4

. SK121 , ,
[4]. PR4
1 ,
[4], n-
C8-C20 [14].
n- C12-C29.

8 C22C34 n-. n-
C16-C18,
[4].
,
, ,

,
[15].

, Rhodococcus
. ,
, ,
[16].

.
Rhodococcus ,
,
. ,
,
, ,
[16].
-

Rhodococcus
.

..

. :
09-04-12231-_, 10-05-00681-, -1901.2010.5.

1. .., .., .., ..,
.., ..

. 2008. . , 3, . 333340.
2. Van Beilen J.B., Li Z., Duetz W.A., Smits T.H.M., Witholt B. Diversity of

Alkane Hydroxylase Systems in the Environment. Oil & Gas Science and
Technology. 2003, V. 58, No. 4, 427-440.

3. Van Beilen J.B., Panke S., Lucchini S., Franchini A.G., Rthlisberger M.,

Witholt B. Analysis of Pseudomonas putida Alkane Degradation Gene


Clusters and Flanking Insertion Sequences: Evolution and Regulation of
the Alk-Genes. Microbiology. 2001, V. 147, 1621-1630.
4. Alvarez H. M. Biology of Rhodococcus / H. M. Alvarez //

Microbiology Monographs : London, New York, Heidelberg,


Dordrecht: Springer, 2010. P. 1-361
5. Sambrook J. Molecular cloning: a laboratory manual / J. Sambrook, E. F.

Fritsch, T. Maniatis. Cold Spring Harbor : Cold Spring Harbor


Laboratory Press. 1989. 1659 p.
6. Construction and fine mapping for recombinant plasmids containing the

rrnB ribosomal RNA operon of E. coli / J. Brosius [et al.] // Plasmid.


1981. V. 6. P. 112-118.
7.

, 16S /
. ., . . , . . [ .] //
. 1999. 6. . 619-624.
8. Development and application of PCR primers for monitoring alkane-

degrading bacteria in seawater microcosm during crude oil degradation


process / K. Sei [et al.] // J. Chem. Eng. Jap. 2003. V. 36, No. 10. P.
1185-1193.
9. Physiological adaptations involved in alkane assimilation at a low

temperature by Rhodococcus sp. strain Q15 / L. G. Whyte [et al.] // App.


Environ. Microbiol. 1999. V. 65, No. 7. P. 2961-2968.
10. Identification of an amino acid position that determines the substrate

range of integral membrane alkane hydroxylases / J. B. Van Beilen [et al.]


// J. Bacteriol. 2005. No. 187. P. 8591.

11. Sequence

analysis of three plasmids harboured in Rhodococcus

erythropolis strain PR4 / M. Sekine [et al.] // Environ. Microbiol. 2006.


V. 8, No. 2. P. 334-346.
12. L. G. Whyte, T. H. M. Smits, D. Labb, B. Witholt, C. W. Greer, and J.

B. van Beilen. Gene Cloning and Characterization of Multiple Alkane


Hydroxylase Systems in Rhodococcus Strains Q15 and NRRL B-16531.
Applied and environmental microbiology, Dec. 2002, p. 59335942.
13. Diversity of alkane hydroxylase systems in the environment / J. B. Van

Beilen [et al.] // OGST. 2003. V. 58, No. 4. P. 427-440.


14. Construction of bacterial consortia that degrade arabian light crude oil / S.

Komukai-Nakamura [et al.] // J. Ferm. Eng. 1996. V. 82, No. 6. P.


570-574.
15. Larkin M. J. Biodegradation and Rhodococcus masters of catabolic

versatility / M. J. Larkin, L. A. Kulakov, C. C. R. Allen // Biotechnology.


2005. V. 16. P. 282290.
16. Jones A. Genus II. Rhodococcus (Zopf 1891) / A. Jones, M. Goodfellow.

Bergeys Manual of Systematic Bacteriology. Berlin : Springer, 2010.


V. 4. P. 1-65.

. 1. ,
. .
, alk-.
: 10 100 .
. 2. 5
8 3
,

alkB-. : 20
100 .

. 1.

. 2.

The complete alk-sequences of Rhodococcus erythropolis of deep-water


bitumen mounds on the bottom of Lake Baikal


, .

664033, - 3.
(3952) 42-54-15, 42-54-05 ()
likhoshvay@mail.ru

664033, - 3.
(3952) 42-54-15, 42-54-05 ()
lomakina@lin.irk.ru

664033, - 3.
(3952) 42-32-80, 42-54-05 ()
yuragal@gmail.com
()
664033, - 3.
(3952) 42-65-04, 42-54-05 ()
grachev@lin.irk.ru