Sawant Shailesh S, Salunke Bipinchandra K, Kim Beom Soo
Department of Chemical Engineering, Chungbuk National University, Cheongju, Chungbuk, 361-763, Republic of Korea.
Curr Microbiol. 2014 Dec;69(6):832-8. doi: 10.1007/s00284-014-0664-y. Epub 2014 Aug 2.
A contaminating bacterium growing along with the stock culture of Saccharophagus degradans ATCC 43961 (Sde 2-40) on marine agar plate was isolated and investigated for its ability to produce polyhydoxyalkonates (PHA). Preliminary screening by Sudan black B and Nile blue A staining indicated positive characteristic of the isolate to produce PHA. The isolate was able to grow and produce PHA in minimal sea salt medium broth. PHA quantification studies with gas chromatographic analyses of the dry cells derived from culture broths revealed accumulation of PHA in bacterial cells. PHA production started after 20 h and increased with cell growth and attained maximum values of 61 % of dry cell weight at 70 h of cultivation. After 70 h, a slight decrease in the level of PHA content was observed. The nature/type of PHA was found to be poly(3-hydroxybutyraye) by Fourier transform-infrared spectroscopy. Microbiological and 16S rRNA gene sequencing analyses suggested that the PHA producing bacterial isolate belongs to Bacillus genera and shows 100 % nucleotide sequence similarity with Bacillus cereus species in GenBank. This study is a first report for ability of Bacillus species to grow in marine sea salt media and produce PHA. The media used for the polymer production was novel in the context of the genus Bacillus and the production of PHA was three-fold higher than Sde 2-40 using same growth medium. This study shows that the contaminant bacteria once properly investigated can be used for advantageous characteristic of metabolites production in place of original cultures.
从与嗜糖降解菌(Saccharophagus degradans)ATCC 43961(Sde 2 - 40)的原种培养物一起在海洋琼脂平板上生长的一种污染细菌中分离出该细菌,并对其产生聚羟基链烷酸酯(PHA)的能力进行了研究。通过苏丹黑B和尼罗蓝A染色进行的初步筛选表明该分离物具有产生PHA的阳性特征。该分离物能够在最低海盐培养基肉汤中生长并产生PHA。对来自培养液的干细胞进行气相色谱分析的PHA定量研究表明细菌细胞中积累了PHA。PHA的产生在20小时后开始,并随着细胞生长而增加,在培养70小时时达到干细胞重量的61%的最大值。70小时后,观察到PHA含量水平略有下降。通过傅里叶变换红外光谱法发现PHA的性质/类型为聚(3 - 羟基丁酸酯)。微生物学和16S rRNA基因测序分析表明,产生PHA的细菌分离物属于芽孢杆菌属,并且在GenBank中与蜡样芽孢杆菌物种显示出100%的核苷酸序列相似性。本研究首次报道了芽孢杆菌属物种在海洋海盐培养基中生长并产生PHA的能力。用于聚合物生产的培养基在芽孢杆菌属的背景下是新颖的,并且使用相同生长培养基时PHA的产量比Sde 2 - 40高三倍。这项研究表明,一旦对污染细菌进行适当研究,就可以利用其产生代谢产物的有利特性来替代原始培养物。