Key Laboratory of Marine Chemistry Theory and Technology, Ministry of Education, Ocean University of China, Qingdao 266100, China.
College of Chemistry &Chemical Engineering, Ocean University of China, Qingdao 266100, China.
Sci Rep. 2017 Jan 4;7:39068. doi: 10.1038/srep39068.
Bacteria can use n-hexadecane as a carbon source, but it remains incompletely understood whether n-hexadecane is transformed into metabolic intermediates prior to cellular uptake or not. We newly isolated a strain identified as Pseudomonas synxantha LSH-7' and conducted chemotaxis experiment of this bacterial strain towards n-hexadecane, hexadecanol and hexadecanoic acid with qualitative assays respectively. Furthermore, we described the identification of extracellular alkane hydroxylase and alcohol dehydrogenase activity; acidification of the culture medium; identification of hexadecanoic acid in the culture medium by the GC-MS analysis; and variation concentration of intracellular n-hexadecane and hexadecanoic acid. A detailed analysis of the experimental data revealed the chemotaxis of this bacterial strain towards n-hexadecane instead of its metabolic intermediates. Our results further suggested that only a fraction of total n-hexadecane followed this path, and alkane hydrolase and hexadecanol dehydrogenase were constitutively expressed when grown in the medium of n-hexadecane. Most strikingly, we quantitatively investigated the concentration of n-hexadecane adsorbed by bacterial chemotaxis. Our findings provided an original insight n-hexadecane might be converted to hexadecanoic acid extracellularly before it was taken up across the cell membrane.
细菌可以将正十六烷作为碳源,但目前仍不完全清楚细菌在摄取细胞之前是否将正十六烷转化为代谢中间产物。我们新分离出一株被鉴定为假单胞菌(Pseudomonas synxantha)LSH-7',并分别用定性分析方法对该菌株进行了正十六烷、十六醇和十六酸的趋化性实验。此外,我们还描述了胞外烷烃羟化酶和醇脱氢酶活性的鉴定、培养基的酸化、GC-MS 分析鉴定培养基中的十六酸以及细胞内正十六烷和十六酸浓度的变化。对实验数据的详细分析表明,该细菌菌株对正十六烷具有趋化性,而不是对其代谢中间产物。我们的结果进一步表明,只有一部分总正十六烷遵循这条途径,并且当在正十六烷培养基中生长时,烷烃水解酶和十六醇脱氢酶是组成型表达的。最引人注目的是,我们定量研究了细菌趋化作用吸附的正十六烷浓度。我们的发现提供了一个新的见解,即正十六烷可能在穿过细胞膜被摄取之前被转化为十六酸并分泌到细胞外。