30 4 2009 4 ENVIRONMENTAL SCIENCE Vol. 30,No. 4 Apr.,2009 1 1,4, 1, 1, 2, 3, 4, 1 3 (11, 100085 ; 2. ( ), 100078 ; 3., 100011 ; 4., 100083) :,52 Q 10,. 1 (EBL0706),, ph,,,ph 619715 2050,5 10 8 CFUΠmL,400 mgπl12 h 98 %.,,. :;; ;16S rdna ; :X172 :A :025023301 (2009) 0421199206 Degradation of Dichlorvos by Rhodobacter sphaeroides ZHAO Kai 1,4, YU Ying 1, J IANG Dan 1, WANG Dong 2, LI Zu2ming 3, HUANG Guo2zhong 4, BAI Zhi2hui 1 (11Research Center for Eco2Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China ; 2. Zhongke2Huoli (Beijing) Biotechnology Corporation, Beijing 100078, China ; 3. Teacher s College of Beijing Union University, Beijing 100011, China ; 4. Civil & Environment Engineering School, University of Science and Technology Beijing, Beijing 100083, China) Abstract : Rhodobacter sphaeroides possesses an extensive range of energy acquiring mechanisms including photosynthesis, lithotrophy, aerobic and anaerobic respiration. It can produce 52aminolevulinic acid, CoQ 10, carotenoids, hydrogen, etc. by fermentation. A Rhodobacter sphaeroides strain was isolated, designated as EBL0706, from soil. The degradation of dichlorvos (DDVP) by the Rhodobacter sphaeroides was investigated. 98 % of DDVP could be degraded in water solution in 12 h when 5 10 8 CFUΠmL Rhodobacter sphaeroides was added to 400 mgπl DDVP solution under ph 6192715 and 20250. This strain could also degrade the DDVP residues on Chinese cabbage leaves effectively. Key words : Rhodobacter sphaeroides ; dichlorvos ; degradation ; 16S rdna ; Chinese cabbage ( Rhodobacter sphaeroides),,.,., ( tetrapyrroles) ( chlorophylls) (heme) B 12 [1 ].,. 52 (ALA) [2,3 ] ;ALA,, [4 ]. Q 10 [5,6 ] ; Q 10,,,,, [7,8 ]. [9 ] [10,11 ]. [12 ].,,2005, [1 ]. (dichlorvos,ddvp) 2,22,,,,,, 011 mgπkg, 0102 mgπkg. :2008206224 ; :2008211230 : (30600082) ; (2008BADA7B01) ;(20061D0502200295, KM200811417006) : (1984),,,. 3,E2mail :zhbai @rcees. ac. cn
1200 30, [13 ]., [1416 ]., 1, : EBL0706,, : CGMCC No1 0645,. 1 1. 1 ( Rhodobacter sphaeroides) EBL0706,LB, 4, 1. (gπl) : 4,1, 019,016,012, 01075, 01012,EDTA 0102, 10 ml, 10 ml, ph 710., (gπl) : 218, 116,0176,0124,0104 ; (gπl) :B 1 1,(VPP) 1, 01016, 1.. 1. 2 100 ml,3,, 32,150 rπmin 24 h,. 5 % 4 L,pH 617,5 LΠmin, 32, 26 h. (CFU). ph ph. 1. 3 PCR 16S rdna 16S rdna 27f (5 2AGAGTT2 TGATCMTGGCTCAG, M = C A) 1492r (5 2TAC2 GGYTACCTTGTTACGACTT, Y = C T), DNA, PCR 16S rdna [17 ]. PCR : 95 5 min, 94 1 min,50 1 min,72 1 min,30,72 10 min. PCR 1 %. 1. 4 16S rdna 16S rdna PCR. NCBI Blast,GenBank,, DNAMAN,. 1. 5 21 h 5 10 8 CFUΠmL,150 ml, 50 ml, ph 618713 715 718 813,121 20 min, 80 % ( ) 400 mgπl. 30 150 rπmin, 2 h ph, ph.,. 21 h 5 10 8 CFUΠmL,50 ml 150 ml, 012m, 5, 80 % 400 600 800 mgπl, (1 0002 000 ). ph 30 150 rπmin, 2 h ph, ph.,. 21 h 1 10 8 2 10 8 5 10 8 CFUΠmL, 50 ml 150 ml,, 80 % 400 mgπl. ph 30 150 rπmin, 2 h ph, ph.,. 21 h 5 10 8 CFUΠmL,150 ml, 50 ml, ph ph,, 80 % 400 mgπl., 150 rπmin, 2 h ph,ph.,. 1. 6 () 80 % 1 000,30 1
4 :1 1201,4,2 h,1 10 8 2 10 8 5 10 8 CFUΠmL,. 24 h 48 h,. 1. 7 GBΠT 500912012003 [18 ] ;2, < 5 %.,100 mgπl. 2 2. 1, 1.,32,, 12 h, 21 h 811 10 9 CFUΠmL, 24 h,. ph, ph 618, 27 h ph 917. ph, ph 915,,., ph.,.,,., ( Rhodobacter ) ( Rhodobacter sphaeroides). 2. 3 16S rdna 16S rdna GenBank,: EU263643. GenBank,> 97 % Rhodobacter, Rhodobacter sphaeroides ATCC 17025, 99 %. 2. 1 Fig. 1 Time course of Rhodobacter sphaeroides growth 2. 2,,,,018112m,,,.,,., 2 16S rdna Fig. 2 Phylogenetic tree based on 16S rdna sequence analysis of Rhodobacter sphaeroides 2. 4 ph, ph,
1202 30 600 800 mgπl, 12 h, ph 515,,. Schramm [19 ], : C 4 H 7 Cl 2 O 4 P + 9Π2O 2 4CO 2 + H 2 O + 2Cl - + PO 3-4 + 5H + (1) (1),1 mol 2 mol 1 mol,5 mol H +.,, 0102 molπlnaoh,ph, NaOH.,NaOH ( ) 5,,,. ph, 3. 2 h 1 ph,,ph. 6 h 12 h, ph 715 619,. ph,, ph,, ph. 7715 %,24 h 9918 %9910 % 9511 %.,. 4 Fig. 4 Degradation of DDVP with different concentration in water solution by Rhodobacter sphaeroides 2. 6 400 mgπl 5.,,. 1 10 8 CFUΠmL,18 h,,. 3 ph Fig. 3 Effect of ph on the degradation of DDVP by Rhodobacter sphaeroides 2. 5 (5 10 8 CFUΠmL) 4.,,400600 800 mgπl 12 h 9814 %8911 % 5 Fig. 5 Degradation of DDVP in water solution by different Rhodobacter sphaeroides addition 2. 7,400 mgπl 6.,50
4 :1 1203,,2 h 77 %. 2050 12 h 98 %,15 55,12 h 97 %95 %.. 1 Table 1 Degradation of DDVP in Chinese cabbage by ΠCFU ml - 1 Rhodobacter sphaeroides Πmg kg - 1 1 h 24 h 48 h 0 3163 0121 0127 0102 01051 01005 1 10 8-01051 01005 01019 01003 2 10 8-01021 01002 01011 01001 5 10 8-01015 01001 01011 01002 6 Fig. 6 Effect of temperature on the degradation of DDVP by Rhodobacter sphaeroides 2. 8,,,, 1.,, 24 h,(011 mgπkg), 5 10 8 CFUΠmL, (0102 mgπkg). 48 h,,., 3 d [20 ],,,24 h 90 %,48 h, 7 d.,,,. Zhang [21 ],,. 2. 9.,,5 10 8 CFUΠmL 300 mgπl 48 h 57 %,600 mgπl48 h 44 %..,. 3 (1) EBL0706 32 21 h 811 10 9 CFUΠmL. 2050,pH 617718,,. (2),,. : [ 1 ] http :ΠΠmmg. uth. tmc. eduπsphaeroidesπintroductionπintroduction. html [ 2 ] Sasaki K, Watanabe M, Nishio N. Inhibition of 52aminolevulinic acid (ALA) dehydratase by undissociated levulinic acid during ALA extracellular formation by Rhodobacter sphaeroides [ J ]. Biotechnol Letters, 1997, 19 (5) : 4212424. [ 3 ],. Rhodobacter sphaeroides 52[J ]., 2005, 11(5) : 6112613. [ 4 ],,,. 52 [J ]., 2003, 39 (3) : 1852192. [ 5 ] Yen H W, Chiu C H. The influences of aerobic2dark and anaerobic2 light cultivation on CoQ 10 production by Rhodobacter sphaeroides in the submerged fermenter[j ]. Enzyme Microbial Technol, 2007, 41 (5) : 6002604. [ 6 ],,. Q 10 [J ]., 2006, 13(3) : 2272232.
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