Benjamin Sailas, Kamimura Naofumi, Takahashi Kenji, Masai Eiji
Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940 2188, Japan; Enzyme Technology Laboratory, School of Biosciences, University of Calicut, Kerala 673635, India.
Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940 2188, Japan.
Ecotoxicol Environ Saf. 2016 Dec;134P1:172-178. doi: 10.1016/j.ecoenv.2016.08.028. Epub 2016 Sep 10.
This study provides physical and analytical evidences for the efficient utilization of most of the commercially available phthalate diesters by Achromobacter denitrificans SP1, coupled with the demonstration of a plausible degradation pathway. We tested 17 phthalate diesters [viz., ditridecyl phthalate, diisodecyl phthalate (DIDP), di(2-ethylhexyl)phthalate (DEHP), di-n-octyl phthalate (DOP), bis(2-ethylhexyl)isophthalate (BEIP), dihexyl phthalate (DHP), dibutyl phthalate (DBP), dicyclohexyl phthalate (DCHP), diphenyl phthalate (DPP), benzyl butyl phthalate (BBP), diamyl phthalate (DAmP), diisobutyl phthalate, dipropyl phthalate, dially phthalate (DAlP), diethyl phthalate, diethyl terephthalate and dimethyl phthalate (DMP)], and their major degradation products for the degradation efficiency of A. denitrificans SP1 in Wx medium. It efficiently utilized 16 phthalate diesters (except DAlP), and showed general preference toward phthalate diesters with longer side chains (utilized 10mM in 48h) than those with shorter side chains and with cyclic structures (utilized ~5mM in 48h) accompanied by a sharp decline of pH to ~5 from initial 7. In a detailed study, about 37mM (15g/L) DEHP was utilized in 48h. Moreover, A. denitrificans SP1 produced reddish-pink pigment when DIDP, DEHP, DOP, DHP, DBP, DIBP, BBP, DAmP, DCHP, DPP or DMP was supplied in the medium. From the available evidences, it seems that its putative phthalate diester degradation pathway contains the following check points: phthalate diesters, phthalate monoesters, phthalate (4,5-dioxygenase), protocatechuate (3,4-dioxygenase), and TCA cycle. Nonspecificity toward utilization of phthalate diesters, especially with greater specificity to phthalate diesters having longer side chain, and the characteristic sticky reddish-pink (or colorless) cell clump formation in the presence of phthalate diesters makes A. denitrificans SP1 a very attractive candidate to be employed as an efficient biofactory in waste water treatment processes.
本研究为反硝化无色杆菌SP1对大多数市售邻苯二甲酸二酯的高效利用提供了物理和分析证据,并证明了一条合理的降解途径。我们测试了17种邻苯二甲酸二酯[即邻苯二甲酸二正十三酯、邻苯二甲酸二异癸酯(DIDP)、邻苯二甲酸二(2-乙基己基)酯(DEHP)、邻苯二甲酸二正辛酯(DOP)、间苯二甲酸双(2-乙基己基)酯(BEIP)、邻苯二甲酸二己酯(DHP)、邻苯二甲酸二丁酯(DBP)、邻苯二甲酸二环己酯(DCHP)、邻苯二甲酸二苯酯(DPP)、邻苯二甲酸苄基丁酯(BBP)、邻苯二甲酸二戊酯(DAmP)、邻苯二甲酸二异丁酯、邻苯二甲酸二丙酯、邻苯二甲酸二烯丙酯(DAlP)、邻苯二甲酸二乙酯、对苯二甲酸二乙酯和邻苯二甲酸二甲酯(DMP)]及其主要降解产物在Wx培养基中对反硝化无色杆菌SP1的降解效率。它能有效利用16种邻苯二甲酸二酯(除DAlP外),并且通常更倾向于利用侧链较长的邻苯二甲酸二酯(48小时内利用约10mM),而不是侧链较短和具有环状结构的邻苯二甲酸二酯(48小时内利用约5mM),同时pH值从初始的7急剧下降到约5。在一项详细研究中,48小时内约37mM(约15g/L)的DEHP被利用。此外,当培养基中提供DIDP、DEHP、DOP、DHP、DBP、DIBP、BBP、DAmP、DCHP、DPP或DMP时,反硝化无色杆菌SP1会产生红粉色色素。从现有证据来看,其假定的邻苯二甲酸二酯降解途径似乎包含以下关键点:邻苯二甲酸二酯、邻苯二甲酸单酯、邻苯二甲酸(4,5-双加氧酶)、原儿茶酸(3,4-双加氧酶)和三羧酸循环。对邻苯二甲酸二酯利用的非特异性,特别是对侧链较长的邻苯二甲酸二酯具有更高的特异性,以及在邻苯二甲酸二酯存在下形成特征性的粘性红粉色(或无色)细胞团,使得反硝化无色杆菌SP1成为废水处理过程中极具吸引力的高效生物工厂候选菌株。