Gan Zhixiong, Zhang Guang, Mo Xiaoyan, Chen Guoqiang, Wu Qiong
School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, China.
Wei Sheng Wu Xue Bao. 2003 Dec;43(6):809-12.
Aeromonas hydrophila WQ isolated from lake water was found to be able to synthesize polyhydroxyalkanoates (PHA) copolymer consisting of 3-hydroxybutyrate (HB) and 3-hydroxyhexanoate (HHx) (PHBHHx). Lauric acid was found to be the most suitable carbon source for cell growth and PHBHHx accumulation. The bacteria accumulated 49% PHBHHx containing 6% HHx in terms of cell dry weight when grown on lauric acid for 72 h. 42% PHBHHx consisting of 14% HHx was obtained with 5 g/L glucose and 10 g/L lauric acid as co-substrate. Higher glucose concentration greatly reduced the cell concentration and PHA content. The PHA biosynthesis genes from A. hydrophila WQ was successfully cloned using a two-step PCR cloning strategy based on PHA biosynthesis genes organization of Aeromonas caviae. A. hydrophila WQ and A. caviae shared high identities in the PHA gene loci, namely, ORF1, phaC and phaJ had 100%, 97% and 97.5% identities respectively. PHA synthases of A. caviae and A. hydrophila were proposed to contain type IV PHA synthases which are different compared with type I PHA synthases on the substrate specificity and location arrangement of PHA metabolic genes.
从湖水中分离出的嗜水气单胞菌WQ被发现能够合成由3-羟基丁酸(HB)和3-羟基己酸(HHx)组成的聚羟基脂肪酸酯(PHA)共聚物(PHBHHx)。发现月桂酸是细胞生长和PHBHHx积累的最合适碳源。当在月桂酸上生长72小时时,细菌以细胞干重计积累了49%含6% HHx的PHBHHx。以5 g/L葡萄糖和10 g/L月桂酸作为共底物时,获得了由14% HHx组成的42% PHBHHx。较高的葡萄糖浓度极大地降低了细胞浓度和PHA含量。基于豚鼠气单胞菌的PHA生物合成基因组织,采用两步PCR克隆策略成功克隆了嗜水气单胞菌WQ的PHA生物合成基因。嗜水气单胞菌WQ和豚鼠气单胞菌在PHA基因位点上具有高度同源性,即ORF1、phaC和phaJ的同源性分别为100%、97%和97.5%。豚鼠气单胞菌和嗜水气单胞菌的PHA合酶被认为含有IV型PHA合酶,这与I型PHA合酶在PHA代谢基因的底物特异性和位置排列上有所不同。