热带海洋学报 JOURNAL OF TROPICAL OCEANOGRAPHY 2010 年第 29 卷第 4 期 :71 75 海洋生物学 http://jto.scsio.ac.cn; http://www.jto.ac.cn Bukholderia sp. DA2 王亚丽 1,2, 殷波 1, 洪义国 1, 顾继东 1 (1., 510301; 2., 510110) : (dimethyl phthalate, DMP),, Bukholderia. sp. DA2 DMP (Dimethyl phthalate, DMT) (Dimethyl dimethyl isophthalate, DMI) 2 B. sp. DA2 8d 100mg L 1 DMT, 15d 400mg L 1 DMP 200mg L 1 DMI,,, B. sp.da2 DMT DMP,,,, : ; ; ; ; 中图分类号 : X172 文献标识码 : A 文章编号 : 1009-5470(2010)04-0071-05 Biodegradation of dimethyl phthalate by Burkholderia sp. DA2 isolated from marine sediments in the South China Sea WANG Ya-li 1,2, YIN Bo 1, HONG Yi-guo 1, GU Ji-dong 1 (1. Key laboratory of Tropical Marine Environment Dynamic, South China Sea Institute of Oceanography, CAS, Guangzhou 510301, China; 2. Guangdong Museum, Guangzhou 510110, China) Abstract: Burkholderia sp. DA2 strain isolated from marine sediments in the South China Sea was capable of utilizing dimethyl phthalate (DMP) as the sole source of carbon and energy for biodegradation of DMP, DMT (dimethyl phthalate) and DMI (dimethyl isophthalate). During the transformation of DMP (400mg L 1 ), its corresponding intermediates were identified as mono-methyl phthalate (MMP) and phthalate acid (PA) by HPLC analysis. By the 9th day, the DMP concentration was undetectable in the culture, corresponding to a DMP decrease; MMP accumulated to 38.9 mg/l during eight days, at the same time amount of PA was also produced during the first 12 days of degradation; this PA was completely degraded after 15 days. During the transformation of DMT (100mg L 1 ), 80% DMT was transferred into MMT in one day, but the MMT could not degraded rapidly and accumulated to 80.2mg L 1, and completely mineralized during eight days. During the DMI (200mg L 1 ) degradation, the DMI was transferred into MMI after six days, but the MMI could not further mineralize. Maybe the compounds chemical structures infect the activity of the enzyme degradation. The biodegradation pathway of DMP, DMT and DMI was proposed as DMP\DMT\DMI MMP\MMT\MMI PA\TA\IA... CO 2 +H 2 O. Key words: DMP; DMI; DMT; biodegradation; marine microorganism (phthalic acid esters, PAEs), PAEs [1] PAEs, PAEs, PAEs,,,, PAEs, 收稿日期 2008-04-10; 修订日期 2008-06-06 基金项目 (06301430); (30800032) 作者简介 (1978 ),,,,
72 Vol. 29, No. 4 / Jul., 2010 (DMP), PAEs (dimethyl isophthalate, DMI) (dimethyl terephthalate, DMT), DMP 2 DMP, (PVC), 20% 30%,,,,,,,, [2] DMP [3], DMP [4], [5 6] DMP [7], DMP [8 9] (Bukholderia cepacia DA2), 1 1.1 (DMP) (DMI) (DMT) (MMP) (PA), (MMI) (IA), (MMT) (TA), Aldrich Chemicals, 99%, 1.2 2002 5 14 1340m 20 1.3 (mg L 1 ): (NH 4 ) 2 SO 4 1000, KH 2 PO 4 200, K 2 HPO 4 800, MgSO 4 7H 2 O 500, FeSO 4 10, CaCl 2 50, NaCl 500, ph 7.0 0.1 : 15g L 1, 0.2μm : 250mL 150mL, DMP (400mg L 1 ) 0.5g, (30.00.5, 150r min 1 ) 6d 1, 1mL 140mL (DMP400mg L 1 ), 4 : DMP,, (30 0.5) 3 1.4, LB,, 1mL 4000 g 5min, 3, DMP (400mg L 1 ) DMI(200mg L 1 ) DMT(100mg L 1 ),, ( 20 ) 1.5 0.2μm, (HPLC) HP1100, Hypersil ODS C 8 (4.0mm 125mm), ( 0.01mol L 1 )(40:60 ), 25min, 1.0mL min 1, 30, 20μl,, DMP DMT(240nm), DMI(236nm), 3, (600nm), 2 2.1 DMP, 1 DMP,,,, 16S rdna Burkholderia sp.da2( ) 2.2 (DMP) Burkholderia sp.da2 (DMP), 6d,,, 8, DMP ( 1), OD 600 0.274 HPLC, DMP,, (MMP, r.t.19.636min), (PA,r.t.16.636min)( 2)
: Bukholderia sp. DA2 73, 8 1d MMT, 0.075 1 Burkholderia sp.da2 DMP Fig. 1 Relationship between DMP removal and Burkholderia sp. DA2 biomass 3 Burkholderia sp.da2 DMT Fig. 3 Relationship between DMT removal and Burkholderia sp.da2 biomass 2 Burkholderia sp.da2 DMP HPLC Fig. 2 DMP and its intermediates in degradation with Burkholderia sp. DA2. 4, HPLC, DMT 2, (MMT r.t.11.267min) (TA r.t.4.221min), 6d,, 38.9mg L 1, 6d, 11d, 15d ( 1) 1, 2,, DMP, DMP, (MMP),, CO 2 H 2 0, ph 6.0, MMP PA,,,,, 2.3 (DMT) 3, Burkholderia sp.da2 DMT, MMT, 80.2mg L 1 MMT,, 7d, MMT 4 Burkholderia sp.da2 DMT HPLC Fig. 4 DMT and its intermediates in degradation with Burkholderia sp.da2. Burkholderia sp.da2 DMT, DMT, MMT Burkholderia sp.da2 DMT, MMT, MMT, MMT, TA,, ph, 6.5
74 Vol. 29, No. 4 / Jul., 2010, TA, HPLC ( 4), (TA),, 2.4 (DMI) 5, Burkholderia sp.da2 DMI, 4d DMI(200mg L 1 ) HPLC( 6), (MMI r.t.10.691min), MMI,, ph5.5, MMI, DMI, ph, DMI MMI, DMI,, 5 Burkholderia sp.da2 DMI Fig. 5 Relationship between DMI removal and Burkholderia sp.da2 biomass Burkholderia sp.da2 DMP DMT, DMI,, 2, DMI 2, DMP DMT,,, 6 Burkholderia sp.da2 DMI HPLC Fig. 6 DMI and its intermediates in degradation with Burkholderia sp.da2. 2.5,,, [10 12],, Burkholderia sp.da2,, 3,,, Burkholderia sp.da2 3 1, 1 DMP DMT MMP MMT, PA TA, CO 2 H 2 O DMI, MMI, DMI, MMI, MMI, Burkholderia sp.da2 DMP DMT DMI :
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