Crawford R L, Bromley J W, Perkins-Olson P E
Appl Environ Microbiol. 1979 Mar;37(3):614-8. doi: 10.1128/aem.37.3.614-618.1979.
An aerobic endospore-forming bacterium, tentatively identified as a strain (JJ-lb) of Bacillus macerans, was isolated by enrichment on 4-hydroxybenzoate (4HBA), using as an inoculum soil taken from a 50 degrees C Iadho hot spring. Enzymatic analyses of cells grown on succinate and 4HBA indicated that strain JJ-1b degrades 4HBA by way of the novel protocatechuate (PCA) 2,3-dioxygenase pathway. Purification of the PCA 2,3-dioxygenase by affinity chromatography allowed the first observation of the immediate ring fission product of PCA, namely, 5-carboxy-2-hydroxymuconic semialdehyde (CHMS; labmda max at pH 7.0 = 348 nm). An affinity column fraction was obtained that decarboxylated CHMS to 2-hydroxymuconic semialdehyde (HMS; lambdamax at pH 7.0 = 375 nm). Thus, conversion of PCA to HMS is accomplished in two steps, 2,3-fission of the PCA ring followed by enzymatic decarboxylation of the ring fission product, forming HMS.
从50摄氏度的伊亚多温泉采集土壤作为接种物，通过在4-羟基苯甲酸（4HBA）上富集培养，分离出一种需氧产芽孢细菌，初步鉴定为浸麻芽孢杆菌菌株（JJ-1b）。对在琥珀酸和4HBA上生长的细胞进行酶分析表明，JJ-1b菌株通过新的原儿茶酸（PCA）2,3-双加氧酶途径降解4HBA。通过亲和层析纯化PCA 2,3-双加氧酶，首次观察到PCA的直接环裂解产物，即5-羧基-2-羟基粘康酸半醛（CHMS；pH 7.0时的最大吸收波长=348nm）。获得了一个亲和柱馏分，它将CHMS脱羧为2-羟基粘康酸半醛（HMS；pH 7.0时的最大吸收波长=375nm）。因此，PCA向HMS的转化分两步完成，先是PCA环的2,3-裂解，然后是环裂解产物的酶促脱羧，形成HMS。
