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首页> 外文期刊>Journal of bacteriology >Metabolism of Allylglycine and cis-Crotylglycine by Pseudomonas putida (arvilla) mt-2 Harboring a TOL Plasmid
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Metabolism of Allylglycine and cis-Crotylglycine by Pseudomonas putida (arvilla) mt-2 Harboring a TOL Plasmid

机译：携带TOL质粒的恶臭假单胞菌（arvilla mt-2）对烯丙基甘氨酸和顺式巴豆甘氨酸的代谢

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Spontaneous mutants which acquired the ability to utilize d-allylglycine (d-2-amino-4-pentenoic acid) and dl-cis-crotylglycine (dl-2-amino-cis-4-hexenoic acid) but not l-allylglycine or dl-trans-crotylglycine could be readily isolated from Pseudomonas putida mt-2 (PaM1). Derivative strains of PaM1 putatively cured of the TOL (pWWO) plasmid were incapable of forming mutants able to utilize the amino acids for growth; however, this ability could be regained by conjugative transfer of the TOL (pWWO) plasmid from a wild-type strain of mt-2 or of the TOL (pDK1) plasmid from a related strain of P. putida (HS1), into cured recipients. dl-Allylglycine-grown cells of one spontaneous mutant (PaM1000) extensively oxidized dl-allylglycine and dl-cis-crotylglycine, whereas only a limited oxidation was observed toward l-allylglycine and dl-trans-crotylglycine. Cell extracts prepared from PaM1000 cells contained high levels of 2-keto-4-hydroxyvalerate aldolase and 2-keto-4-pentenoic acid hydratase, the latter enzyme showing higher activity toward 2-keto-cis-4-hexenoic acid than toward the trans isomer. Levels of other enzymes of the TOL degradative pathway, including toluate oxidase, catechol-2,3-oxygenase, 2-hydroxymuconic semialdehyde hydrolase, and 2-hydroxymuconic semialdehyde dehydrogenase, were also found to be elevated after growth on allylglycine. Whole cells of a putative cured strain, PaM3, accumulated 2-keto-4-pentenoic acid from d-allylglycine, which was shown to be rapidly degraded by cell extracts of PaM1000 grown on dl-allylglycine. These same cell extracts were also capable of catalyzing the dehydrogenation of d- but not l-allylglycine and were further found to metabolize the amino acid completely to pyruvate and acetaldehyde. Differential centrifugation of crude cell extracts localized d-allylglycine dehydrogenase activity to membrane fractions. The results are consistent with a catabolic pathway for d-allylglycine and dl-cis-crotylglycine involving the corresponding keto-enoic acids as intermediates, the further metabolism of which is effected by the action of TOL plasmid-encoded enzymes.
机译：具有利用d-烯丙基甘氨酸（d-2-氨基-4-戊烯酸）和dl- 顺式-巴豆基甘氨酸（dl-2-氨基-顺式 -4-己烯酸），而不是l-烯丙基甘氨酸或dl- trans -巴豆基甘氨酸。推定为TOL（pWWO）质粒固化的PaM1衍生菌株不能形成能够利用氨基酸生长的突变体。但是，可以通过野生型mt-2菌株的TOL（pWWO）质粒或来自 P的相关菌株的TOL（pDK1）质粒的共轭转移来恢复这种能力。恶臭（HS1），转移到治愈的收件人中。一个自发突变体（PaM1000）的dl-烯丙基甘氨酸生长的细胞广泛氧化dl-烯丙基甘氨酸和dl- 顺式-巴豆基甘氨酸，而仅观察到有限的向l-烯丙基甘氨酸和dl- 反式的氧化-巴豆甘氨酸。从PaM1000细胞制备的细胞提取物中含有高水平的2-酮-4-羟戊酸醛缩酶和2-酮-4-戊烯酸水合酶，后者对2-酮- cis -4具有更高的活性-己酸比对反式异构体。还发现，在烯丙基甘氨酸上生长后，TOL降解途径的其他酶的水平也升高，包括甲苯磺酸氧化酶，儿茶酚-2,3-加氧酶，2-羟基粘康半醛水解酶和2-羟基粘康半醛脱氢酶。推定的已治愈菌株PaM3的全细胞从d-烯丙基甘氨酸中积累了2-酮基-4-戊烯酸，并被在dl-烯丙基甘氨酸上生长的PaM1000的细胞提取物迅速降解。这些相同的细胞提取物也能够催化d-烯丙基甘氨酸的脱氢反应，但不能催化氨基酸完全代谢为丙酮酸和乙醛。粗细胞提取物的差异离心将d-烯丙基甘氨酸脱氢酶活性定位于膜级分。结果与涉及相应的酮-烯酸作为中间体的d-烯丙基甘氨酸和dl- 顺式-巴豆基甘氨酸的分解代谢途径一致，其进一步代谢受TOL编码质粒的作用影响酶。

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