Degradation of Dichlorvos by Rhodobacter sphaeroides

Transcript

1 ENVIRONMENTAL SCIENCE Vol. 30,No. 4 Apr., ,4, 1, 1, 2, 3, 4, 1 3 (11, ; 2. ( ), ; 3., ; 4., ) :,52 Q 10,. 1 (EBL0706),, ph,,,ph , CFUΠmL,400 mgπl12 h 98 %.,,. :;; ;16S rdna ; :X172 :A : (2009) 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 , China ; 2. Zhongke2Huoli (Beijing) Biotechnology Corporation, Beijing , China ; 3. Teacher s College of Beijing Union University, Beijing , China ; 4. Civil & Environment Engineering School, University of Science and Technology Beijing, Beijing , 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 CFUΠmL Rhodobacter sphaeroides was added to 400 mgπl DDVP solution under ph and 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. : ; : : ( ) ; (2008BADA7B01) ;(20061D , KM ) : (1984),,,. 3,E2mail ac. cn

2 , [13 ]., [1416 ]., 1, : EBL0706,, : CGMCC No1 0645, ( 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, ml,3,, 32,150 rπmin 24 h,. 5 % 4 L,pH 617,5 LΠmin, 32, 26 h. (CFU). ph ph 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 % S rdna 16S rdna PCR. NCBI Blast,GenBank,, DNAMAN, h CFUΠmL,150 ml, 50 ml, ph , min, 80 % ( ) 400 mgπl rπmin, 2 h ph, ph.,. 21 h CFUΠmL,50 ml 150 ml, 012m, 5, 80 % mgπl, ( ). ph rπmin, 2 h ph, ph.,. 21 h CFUΠmL, 50 ml 150 ml,, 80 % 400 mgπl. ph rπmin, 2 h ph, ph.,. 21 h CFUΠmL,150 ml, 50 ml, ph ph,, 80 % 400 mgπl., 150 rπmin, 2 h ph,ph., () 80 % 1 000,30 1

3 4 :1 1201,4,2 h, CFUΠmL,. 24 h 48 h, GBΠT [18 ] ;2, < 5 %.,100 mgπl , 1.,32,, 12 h, 21 h CFUΠmL, 24 h,. ph, ph 618, 27 h ph 917. ph, ph 915,,., ph.,.,,., ( Rhodobacter ) ( Rhodobacter sphaeroides) S rdna 16S rdna GenBank,: EU GenBank,> 97 % Rhodobacter, Rhodobacter sphaeroides ATCC 17025, 99 % 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,

4 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 H + (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 ,. ph,, ph,, ph %,24 h 9918 %9910 % 9511 %.,. 4 Fig. 4 Degradation of DDVP with different concentration in water solution by Rhodobacter sphaeroides mgπl 5.,, CFUΠmL,18 h,,. 3 ph Fig. 3 Effect of ph on the degradation of DDVP by Rhodobacter sphaeroides 2. 5 ( CFUΠmL) 4.,, 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

5 4 :1 1203,,2 h 77 % 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 h 24 h 48 h Fig. 6 Effect of temperature on the degradation of DDVP by Rhodobacter sphaeroides 2. 8,,,, 1.,, 24 h,(011 mgπkg), CFUΠmL, (0102 mgπkg). 48 h,,., 3 d [20 ],,,24 h 90 %,48 h, 7 d.,,,. Zhang [21 ],, ,, CFUΠmL 300 mgπl 48 h 57 %,600 mgπl48 h 44 %..,. 3 (1) EBL h CFUΠmL. 2050,pH ,,. (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) : [ 3 ],. Rhodobacter sphaeroides 52[J ]., 2005, 11(5) : [ 4 ],,,. 52 [J ]., 2003, 39 (3) : [ 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) : [ 6 ],,. Q 10 [J ]., 2006, 13(3) :

6 [ 7 ],,,. Q 10 [J ]., 2006, 8 (1) : [ 8 ]. Q 10 [J ]., 2006,22 (4) : [ 9 ] Chen D, Han Y, Gu Z. Application of statistical methodology to the optimization of fermentative medium for carotenoids production by Rhodobacter sphaeroides[j ]. Process Biochemistry, 2006, 41 (8) : [10 ] Koku H, Eroglu I, Gunduz U, et al. Aspects of the metabolism of hydrogen production by Rhodobacter sphaeroides [J ]. International J Hydrogen Energy, 2002, 27(11212) : [11 ] Fang H H P, Zhu H, Zhang T. Phototrophic hydrogen production from glucose by pure and co2cultures of Clostridium butyricum and Rhodobacter sphaeroides[j ]. International J Hydrogen Energy, 2006, 31(15) : [12 ]. [ P]. : ZL , [13 ] Zhang J L, Qiao C L. Novel approaches for remediation of pesticide pollutants[j ]. International Journal of Environment Pollution, 2002, 18( 5) : [14 ],. [J ].,2006, 29 (4) : [15 ],,,. DDB21 [J ]., 2007, 26 (2) : [16 ] Oncescu T, Oancea P, Enache M, et al. Halophilic bacteria are able to decontaminate dichlorvos, a pesticide, from saline environments[j ]. Central European Journal of Biology,2007, 2(4) : [17 ] Li Z, Bai Z, Zhang B, et al. Newly isolated Bacillus gibsonii S22 capable of using sugar beet pulp for alkaline pectinase production[j ]. World Journal of Microbiology and Biotechnology, 2005, 21 : [18 ],,,. [J ].,2008,29 (4) : [19 ] Schramm J D, Hua I. Ultrasonic Irradiation of Dichlorvos : Decomposition Mechanism[J ]. Water Research, 2001, 35 (3) : [20 ] Australian Pesticides & Veterinary Medicines Authority. Dichlorvos toxicology assessment [ M ]. Australia : Australian Pesticides & Veterinary Medicines Authority, [21 ] Zhang B, Zhang H, Jin B, et al. Effect of cypermethrin insecticide on the microbial community in cucumber phyllosphere[j ]. Journal of Environmental Sciences, 2008, 20 (11) :