Benzene-free synthesis of adipic acid
Benzene-free synthesis of adipic acid
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DOI：
10.1021/bp010179x
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发表时间：
2002-03-01
影响因子：
2.9
通讯作者：
Frost, JW
中科院分区：
文献类型：
--
作者：
Niu, W;Draths, KM;Frost, JW
构建并评价了在流加发酵罐条件下以D-葡萄糖为底物合成顺，顺-粘康酸的大肠杆菌菌株。还检查了在所得发酵液中的顺式，顺式-粘康酸的化学氢化。从葡萄糖生物催化合成己二酸消除了工业己二酸生产的两个环境问题：使用致癌的苯和苯衍生的化学品作为原料和生成一氧化二氮作为硝酸催化氧化的副产物。虽然已经开发了替代催化合成，消除了硝酸的使用，但大多数仍然依赖于石油衍生的苯作为最终原料。在本研究中，E. coliWN 1/pWN2.248在补料分批发酵罐条件下培养48 h，以葡萄糖为原料合成顺，顺粘康酸36.8g/L，产率22%（mol/mol）。优化微生物顺，顺粘康酸合成需要表达三种在大肠杆菌中不常见的酶。杆菌将编码DHS脱氢酶的肺炎克雷伯氏菌aroZ基因的两个拷贝插入E. coli染色体，而K.编码PCA脱羧酶的pneumoniae aroY基因和编码儿茶酚1，2-双加氧酶的醋酸钙不动杆菌catA基因从染色体外质粒表达。在WN 1/pWN2.248的补料分批培养完成后，从肉汤中取出细胞，用活性炭处理，随后过滤以除去可溶性蛋白。在3400 kPa的H-2压力下，在环境温度下，用10%Pt/碳（5%mol/mol）氢化所得溶液2.5小时，得到97%（mol/mol）的顺式，顺式-粘康酸转化为己二酸。
Strains of Escherichia coli were constructed and evaluated that synthesized cis,cis-muconic acid from D-glucose under fed-batch fermentor conditions. Chemical hydrogenation of the cis,cis-muconic acid in the resulting fermentation broth has also been examined. Biocatalytic synthesis of adipic acid from glucose eliminates two environmental concerns characteristic of industrial adipic acid manufacture: use of carcinogenic benzene and benzene-derived chemicals as feedstocks and generation of nitrous oxide as a byproduct of a nitric acid catalyzed oxidation. While alternative catalytic syntheses that eliminate the use of nitric acid have been developed, most continue to rely on petroleum-derived benzene as the ultimate feedstock. In this study, E. coli WN1/pWN2.248 was developed that synthesized 36.8 g/L of cis,cis-muconic acid in 22% (mol/mol) yield from glucose after 48 h of culturing under fed-batch fermentor conditions. Optimization of microbial cis,cis-muconic acid synthesis required expression of three enzymes not typically found in E. coli. Two copies of the Klebsiella pneumoniae aroZ gene encoding DHS dehydratase were inserted into the E. coli chromosome, while the K. pneumoniae aroY gene encoding PCA decarboxylase and the Acinetobacter calcoaceticus catA gene encoding catechol 1,2-dioxygenase were expressed from an extrachromosomal plasmid. After fed-batch culturing of WN1/pWN2.248 was complete, the cells were removed from the broth, which was treated with activated charcoal and subsequently filtered to remove soluble protein. Hydrogenation of the resulting solution with 10% Pt on carbon (5% mol/mol) at 3400 kPa of H-2 pressure for 2.5 h at ambient temperature afforded a 97% (mol/mol) conversion of cis,cis-muconic acid into adipic acid.