第3 4 卷 第 4 期 上海 师范大学学报(自然科学版) Vol.34,No.4
2 0 0 5 年 1 2 月 Journal of Shanghai N orm al U n iversity( N atural Sciences) 2005,Dec.
E ffe c t o f G D O I in te r r u p tio n
in degradation of arom atic com pounds
ZHAO Yu
1,2
, G U O Lu-sheng
1
,XU Ya-tong
2
,ZHOU Xia
1
, L U Y i-ton g
3
(1.College of L ife and E n viron m ent S cien ce, Shanghai N orm al U n iversity, Shanghai, 200234, C hina;
2.C ollege of E nvironm ental Science and R esources, E ast C hina N orm al U niversity, Shanghai, 200062, C hina;
3.D epartm ent of E nvironm ent and R esources, Shanghai Jiaotong U niversity,Shanghai。201101)
A bstract:
Pseudom onas alcaligenes N C IB 9867 (strain P25X ), a soilbacterium , is capable of degrading arom atic
h ydrocarb on viathegentisate path w ay. It h as been estab lish ed that the gentisate pathw ay harbors isozym es for the
g en tisa te m etab olism - on e set is c on stitu tive ly ex p re sse d w h ere a s the other set is strictly in d u cib le. G en tisa te 1 , 2 -
dioxygenase (G D O ) is the key enzym e of the gentisate pathw ay. Itcatalyzes the fission of the gentisate arom atic ring
to yie ld m aleylpyruvate, which then yield s T C A cy c le in term e d iates. T o d ate , th e ge n e s for th e co n stitutively ex-
p ressed gentisate 1 , 2 -dioxygenase an d its dow nstream en zym es have been clo n e d . W e h av e o b tain ed a d eriva tive
strain of P 2 5 X , d esignated as SN Z28, w hich had its constitutive copy of the gentisate 1 , 2-dioxygen ase genes inter-
ru p ted by a streptom ycin /spectinom ycin resistance gen e cassette. In this study, the proteom e profiles of SN Z28,
grown in the m inimalmedium withorwithout the arom atic in ducer, gentisate, w ere com pared . C rude extractsof
SN Z28 cells w ere subjected to tw o-d im ensional polyacrylam aide gel electrop horesis ( P AGE) and stained with Coom-
assie b lu e a n d s ilv e r s taining. 9 de novo protein spots and 7 up - regulated protein spots w ere observedinthe extracts
o b tain ed u n d e r g en tisa te in d u ctio n co n d ition. The results obtained provide us clues for further investiga tion o f induc-
ib le G D O e n zym e .
Keywords:
proteom ic; gentisate; Pseudom onas alcaligenes
C L C num ber:
X172
D ocum ent code:
A
Article ID:
1000-5137(2005)04-0077-06
R e ce iv ed d a te : 2 0 0 4 -1 1 -2 5
B iography: ZH A O Y u( 1973 - ),m ale, Ph.D candidate, lecturer, C ollege of Life an d E n v iro m en tal Science, Shanghai
N orm al U n iversity.
1 In tro d u c tio n
The term “p ro te o m ic s ” w a s firs t m e n tio n e d in 1 9 9 5 a n d w a s d e fin e d a s th e la rg e - s c a le c h a ra c te riza tio n
o f th e e n tire p ro te in c o m p le m e n t o f a c e ll lin e , tiss u e , o r o rg a n ism
[1 ~2 ]
. T o d a y , tw o d e fin itio n s o f p ro te o m ic s
a re p ro p o s e d . T h e first is th e c la s s ic a l d e fin itio n , re stric tin g th e la rg e - sc ale an aly sis of g en e p ro d u c ts. T h e
s e c o n d a n d m o re in c lu siv e d e fin itio n c o m b in e s p ro te in s tu d ie s w ith a n a ly ses that have a genetic readoutsuch as
m R N A an alysis, gen om ics, and th e yeast tw o - hybrid analysis
[4]
. H ow ever, the goal of proteom ics rem ains
the sam e, i.e., to obtain a m ore global and integrated view of biology by stud y in g a ll th e p ro te in o f a c e ll
[5]
.
Pseudom onas alcaligenes N C IB 9867 ( strain P25X ) is a soil bacterium that is capable of degrading xy-
le n o ls a n d c re s o ls v ia th e g e n tisa te p a th w a y . It h a s b e e n e sta b lis h e d th a t P 25 X has isozym es for the m on o -