Vomberg A, Klinner U
Lehr-und Forschungsgebiet Angewandte Mikrobiologie, Institut für Biologie IV (Mikrobiologie), Worringer Weg, Aachen, Germany.
J Appl Microbiol. 2000 Aug;89(2):339-48. doi: 10.1046/j.1365-2672.2000.01121.x.
Fifty-four bacterial strains belonging to 37 species were tested for their ability to assimilate short chain and/or medium chain liquid n-alkanes. A gene probe derived from the alkB gene of Pseudomonas oleovorans ATCC 29347 was utilized in hybridization experiments. Results of Southern hybridization of PCR-amplificates were compared with those of colony hybridization and dot blot hybridization. Strongest signals were received only from Gram-negative bacteria growing solely with short n-alkanes (C10). Hybridization results with soil isolates growing with n-alkanes of different chain lengths suggested as well that alkB genes seem to be widespread only in solely short-chain n-alkane-degrading pseudomonads. PCR products of Rhodococcus sp., Nocardioides sp., Gordona sp. and Sphingomonas sp. growing additionally or solely with medium-chain n-alkane as hexadecane had only few sequence identity with alkB though hybridizing with the gene probe. The derived amino acid sequence of the alkB-amplificate of Pseudomonas aureofaciens showed high homology (95%) with AlkB from Ps. oleovorans. alkB gene disruptants were not able to grow with decane.
对属于37个物种的54株细菌进行了同化短链和/或中链液态正构烷烃能力的测试。使用源自食油假单胞菌ATCC 29347的alkB基因的基因探针进行杂交实验。将PCR扩增产物的Southern杂交结果与菌落杂交和斑点印迹杂交结果进行比较。仅从仅以短链正构烷烃(C10)生长的革兰氏阴性细菌中获得最强信号。对以不同链长正构烷烃生长的土壤分离物的杂交结果也表明,alkB基因似乎仅在仅降解短链正构烷烃的假单胞菌中广泛存在。另外或仅以中链正构烷烃十六烷生长的红球菌属、诺卡氏菌属、戈登氏菌属和鞘氨醇单胞菌属的PCR产物,虽然与基因探针杂交,但与alkB的序列同一性很少。金黄色假单胞菌alkB扩增产物的推导氨基酸序列与食油假单胞菌的AlkB具有高度同源性(95%)。alkB基因破坏体不能以癸烷生长。