Department of Microbiology, Maulana Azad College, 8 Rafi Ahmed Kidwai Road, Kolkata, 700013, India,
World J Microbiol Biotechnol. 2014 May;30(5):1575-82. doi: 10.1007/s11274-013-1580-1. Epub 2013 Dec 14.
Cells of Azotobacter chroococcum MAL-201 (MTCC 3853) are capable of accumulating the intracellular poly(3-hydroxybutyric acid) [P(3HB)], accounting for 65-71 % of its cell dry weight and also capable of synthesizing the enzyme alkaline phosphatase (APase), when grown in glucose and tricalcium phosphate containing nitrogen-free modified Stockdale medium. The concentration of insoluble phosphate in broth medium was optimized as 0.25 % (w/v) for growth and biosynthesis of APase. However, the suboptimal concentration of phosphate (0.1 %, w/v) appeared as the best suited for accumulation of P(3HB) by the strain. The significant differences were observed in biosynthesis of polymer and APase enzyme under variable phosphate concentrations. Glucose, 3.0 % (w/v) was recorded as the optimum concentration for all of the three parameters. The continuation of APase biosynthesis was observed during the period of significant decline in the cellular content of the polymer in the late phase of growth. In order to study the role of P(3HB), the rate of autodigestion of biopolymer and phosphate solubilization rate (k, mineralization constant) were determined in carbon-free medium under batch cultivation process and the parameters were found to be positively correlated. The maximum phosphate solubilization rate (k = 0.0154) by the strain MAL-201 timed at the 10th hour of incubation when the rate of polymer degradation concomitantly attained its peak corresponding to 87 mg/l/h and then declined gradually. Only a negligible amount of residual polymer remained undigested. These data strongly support the functional role of P(3HB) in response to multinutritional stress condition.
固氮菌 MAL-201(MTCC 3853)细胞能够积累细胞内聚(3-羟基丁酸)[P(3HB)],占其细胞干重的 65-71%,当在含有无氮改良斯托克代尔培养基的葡萄糖和磷酸三钙中生长时,还能够合成碱性磷酸酶(APase)。在发酵培养基中,不溶性磷酸盐的浓度优化为 0.25%(w/v),以促进 APase 的生长和生物合成。然而,磷酸盐的亚最佳浓度(0.1%,w/v)似乎最适合该菌株积累 P(3HB)。在不同磷酸盐浓度下,聚合物和 APase 酶的生物合成存在显著差异。葡萄糖,3.0%(w/v)被记录为所有三个参数的最佳浓度。在生长后期聚合物细胞含量显著下降期间,观察到 APase 生物合成的持续进行。为了研究 P(3HB)的作用,在分批培养过程中,在无碳培养基中测定了生物聚合物的自降解率和磷酸盐溶解率(k,矿化常数),并发现这些参数呈正相关。当聚合物降解速率同时达到其峰值 87mg/l/h 时,菌株 MAL-201 的最大磷酸盐溶解率(k=0.0154)在培养的第 10 小时达到,然后逐渐下降。只有微不足道的残留聚合物未被消化。这些数据有力地支持了 P(3HB)在应对多营养胁迫条件下的功能作用。