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戊糖乳杆菌、干酪乳杆菌和植物乳杆菌中D-木糖的转运:通过磷酸烯醇丙酮酸:甘露糖磷酸转移酶系统促进扩散机制的证据。

Transport of D-xylose in Lactobacillus pentosus, Lactobacillus casei, and Lactobacillus plantarum: evidence for a mechanism of facilitated diffusion via the phosphoenolpyruvate:mannose phosphotransferase system.

作者信息

Chaillou S, Pouwels P H, Postma P W

机构信息

EC Slater Institute, BioCentrum, University of Amsterdam, 1018 TV Amsterdam, The Netherlands.

出版信息

J Bacteriol. 1999 Aug;181(16):4768-73. doi: 10.1128/JB.181.16.4768-4773.1999.

Abstract

We have identified and characterized the D-xylose transport system of Lactobacillus pentosus. Uptake of D-xylose was not driven by the proton motive force generated by malolactic fermentation and required D-xylose metabolism. The kinetics of D-xylose transport were indicative of a low-affinity facilitated-diffusion system with an apparent K(m) of 8.5 mM and a V(max) of 23 nmol min(-1) mg of dry weight(-1). In two mutants of L. pentosus defective in the phosphoenolpyruvate:mannose phosphotransferase system, growth on D-xylose was absent due to the lack of D-xylose transport. However, transport of the pentose was not totally abolished in a third mutant, which could be complemented after expression of the L. curvatus manB gene encoding the cytoplasmic EIIB(Man) component of the EII(Man) complex. The EII(Man) complex is also involved in D-xylose transport in L. casei ATCC 393 and L. plantarum 80. These two species could transport and metabolize D-xylose after transformation with plasmids which expressed the D-xylose-catabolizing genes of L. pentosus, xylAB. L. casei and L. plantarum mutants resistant to 2-deoxy-D-glucose were defective in EII(Man) activity and were unable to transport D-xylose when transformed with plasmids containing the xylAB genes. Finally, transport of D-xylose was found to be the rate-limiting step in the growth of L. pentosus and of L. plantarum and L. casei ATCC 393 containing plasmids coding for the D-xylose-catabolic enzymes, since the doubling time of these bacteria on D-xylose was proportional to the level of EII(Man) activity.

摘要

我们已经鉴定并表征了戊糖乳杆菌的D-木糖转运系统。D-木糖的摄取不是由苹果酸-乳酸发酵产生的质子动力驱动的,而是需要D-木糖代谢。D-木糖转运的动力学表明这是一个低亲和力的易化扩散系统,其表观K(m)为8.5 mM,V(max)为23 nmol min(-1) mg干重(-1)。在磷酸烯醇丙酮酸:甘露糖磷酸转移酶系统有缺陷的两个戊糖乳杆菌突变体中,由于缺乏D-木糖转运,无法在D-木糖上生长。然而,在第三个突变体中,戊糖的转运并未完全消除,在表达弯曲乳杆菌编码EII(Man)复合物细胞质EIIB(Man)组分的manB基因后,该突变体能够得到互补。EII(Man)复合物也参与干酪乳杆菌ATCC 393和植物乳杆菌80中的D-木糖转运。在用表达戊糖乳杆菌D-木糖分解代谢基因xylAB的质粒转化后,这两个物种能够转运和代谢D-木糖。对2-脱氧-D-葡萄糖具有抗性的干酪乳杆菌和植物乳杆菌突变体的EII(Man)活性有缺陷,在用含有xylAB基因的质粒转化后无法转运D-木糖。最后,发现D-木糖的转运是戊糖乳杆菌以及含有编码D-木糖分解代谢酶质粒的植物乳杆菌和干酪乳杆菌ATCC 393生长的限速步骤,因为这些细菌在D-木糖上的倍增时间与EII(Man)活性水平成正比。

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