Matsushita K, Arents J C, Bader R, Yamada M, Adachi O, Postma P W
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan.
E. C. Slater Instituut, BioCentrum, University of Amsterdam, Amsterdam, The Netherlands.
Microbiology (Reading). 1997 Oct;143 ( Pt 10):3149-3156. doi: 10.1099/00221287-143-10-3149.
Many bacteria can synthesize the cofactor pyrroloquinoline quinone (PQQ), a cofactor of several dehydrogenases, including glucose dehydrogenase (GCD). Among the enteric bacteria, Klebsiella pneumoniae has been shown to contain the genes required for PQQ biosynthesis. Escherichia coli and Salmonella typhimurium were thought to be unable to synthesize PQQ but it has been reported that strain EF260, a derivative of E. coli FB8, can synthesize PQQ after mutation and can oxidize glucose to gluconate via the GCD/PQQ pathway (F. Biville, E. Turlin & F. Gasser, 1991, J Gen Microbiol 137, 1775-1782). We have re-investigated this claim and conclude that it is most likely erroneous. (i) Strain EF260, isolated originally by Biville and coworkers, was unable to synthesize a holo-enzyme GCD unless PQQ was supplied to the growth medium. No GCD activity could be detected in membrane fractions. (ii) The amount of PQQ detected in the growth medium of EF260 was very low and not very different from that found in a medium with its parent strain or in a medium containing no cells. (iii) EF260 cells were unable to produce gluconate from glucose via the PQQ/GCD pathway. (iv) Introduction of a gcd::Cm deletion in EF260, eliminating GCD, did not affect glucose metabolism. This suggested a pathway for glucose metabolism other than the PQQ/GCD pathway. (v) Glucose uptake and metabolism in EF260 involved a low-affinity transport system of unknown identity, followed most likely by phosphorylation via glucokinase. It is concluded that E. coli cannot synthesize PQQ and that it lacks genes required for PQQ biosynthesis.
许多细菌能够合成辅因子吡咯喹啉醌(PQQ),它是包括葡萄糖脱氢酶(GCD)在内的几种脱氢酶的辅因子。在肠道细菌中,肺炎克雷伯菌已被证明含有PQQ生物合成所需的基因。大肠杆菌和鼠伤寒沙门氏菌被认为无法合成PQQ,但据报道,大肠杆菌FB8的衍生物EF260菌株在发生突变后能够合成PQQ,并能通过GCD/PQQ途径将葡萄糖氧化为葡萄糖酸(F. Biville、E. Turlin和F. Gasser,1991年,《普通微生物学杂志》137卷,1775 - 1782页)。我们重新研究了这一说法,得出的结论是它很可能是错误的。(i)最初由Biville及其同事分离得到的EF260菌株,除非向生长培养基中提供PQQ,否则无法合成全酶GCD。在膜组分中未检测到GCD活性。(ii)在EF260的生长培养基中检测到的PQQ量非常低,与在其亲本菌株的培养基或不含细胞的培养基中发现的量没有太大差异。(iii)EF260细胞无法通过PQQ/GCD途径从葡萄糖产生葡萄糖酸。(iv)在EF260中引入gcd::Cm缺失以消除GCD,并不影响葡萄糖代谢。这表明存在一条不同于PQQ/GCD途径的葡萄糖代谢途径。(v)EF260中的葡萄糖摄取和代谢涉及一种身份不明的低亲和力转运系统,随后很可能通过葡萄糖激酶进行磷酸化。结论是大肠杆菌不能合成PQQ,并且它缺乏PQQ生物合成所需的基因。