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利用营养缺陷型突变体分离2,6-二氨基庚二酸的L-型或D-型异构体。

Use of auxotrophic mutants to isolate LL- or DD-isomers of 2,6-diaminopimelic acid.

作者信息

Saleh F, White P J

出版信息

J Gen Microbiol. 1976 Oct;96(2):253-61. doi: 10.1099/00221287-96-2-253.

Abstract

Pseudomonas aeruginosa PAC7 (a mutant deficient in diaminopimelate epimerase), excreted diaminopimelate (solely LL-isomer) after growth in a minimal medium plus lysine with succinate as carbon source. More diaminopimelate was excreted when bacteria were transferred at the end of the exponential phase of growth into fresh minimal medium without lysine but supplemented with pyrivate and additional (NH4)2SO4. The excreted LL-isomer was isolated from the culture filtrate by ion-exchange chromatography and purified by crystallization (1.7 g/9 1 culture). A diaminopimelate-requiring mutant of Bacillus megaterium NCIB7581 grew on LL- and/or meso-diaminopimelate but not on the DD-isomer. This mutant was used to isolate the DD-idomer from a mixture of synthetic LL- and DD-diaminopimelate. It was grown in a minimal medium containing glycerol as carbon source and LL- plus DD-diaminopimelate at a growth-limiting concentration (300 mg l-1); when growth stopped, the DD-diaminopimelate that remained in the culture was isolated and crystallized (1.0 g/II l culture).

摘要

铜绿假单胞菌PAC7(一种二氨基庚二酸差向异构酶缺陷型突变体),在以琥珀酸盐作为碳源的基本培养基中添加赖氨酸培养后,会分泌二氨基庚二酸(仅为LL-异构体)。当细菌在生长指数期结束时转移至不含赖氨酸但添加了丙酮酸盐和额外硫酸铵的新鲜基本培养基中时,会分泌更多的二氨基庚二酸。通过离子交换色谱从培养滤液中分离出分泌的LL-异构体,并通过结晶进行纯化(1.7克/9升培养物)。巨大芽孢杆菌NCIB7581的一种需要二氨基庚二酸的突变体能够在LL-和/或内消旋二氨基庚二酸上生长,但不能在DD-异构体上生长。该突变体用于从合成的LL-和DD-二氨基庚二酸混合物中分离出DD-异构体。它在以甘油作为碳源、含有生长限制浓度(300毫克/升)的LL-和DD-二氨基庚二酸的基本培养基中生长;当生长停止时,分离并结晶培养物中剩余的DD-二氨基庚二酸(1.0克/11升培养物)。

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