Wei Yu-Hong, Chen Wei-Chuan, Huang Chin-Kuei, Wu Ho-Shing, Sun Yi-Ming, Lo Chi-Wei, Janarthanan Om-Murugan
Graduate School of Biotechnology and Bioengineering, Yuan Ze University, Chung-Li, Taoyuan 320, Taiwan; E-Mails:
Int J Mol Sci. 2011 Jan 5;12(1):252-65. doi: 10.3390/ijms12010252.
Polyhydroxyalkanoate (PHA) is a biodegradable material with many potential biomedical applications, including medical implants and drug delivery. This study developed a system for screening production strains in order to optimize PHA production in Cupriavidus taiwanensis 184, 185, 186, 187, 204, 208, 209 and Pseudomona oleovorans ATCC 29347. In this study, Sudan black B staining, Infrared (IR) and Gas Chromatography (GC) analysis indicated that the best strain for PHA synthesis is C. taiwanensis 184, which obtains polyhydroxybutyrate (PHB). Cultivation of C. taiwanensis 184 under a pH of 7.0, at 30 °C, and at an agitation rate of 200 rpm, obtained a PHB content of 10% and PHB production of 0.14 g/L. The carbon and nitrogen types selected for analysis of PHB production by C. taiwanensis 184 were gluconic acid and NH(4)Cl, respectively. Optimal carbon/nitrogen ratio for PHB production was also determined. This study demonstrated a PHB content of 58.81% and a PHB production of 2.44 g/L when the carbon/nitrogen ratio of 8/1 was selected for C. taiwanensis 184. A two-stage fermentation strategy significantly enhanced PHB content and PHB production. Under a two-stage fermentation strategy with nutrient-limited conditions, C. taiwanensis 184 obtained a PHB content of 72% and a PHB concentration of 7 g/L. Finally, experimental results confirmed that optimizing the growth medium and fermentation conditions for cultivating the indigenous C. taiwanensis 184 strain substantially elevated PHB content from 10% to 72% and PHB production from 0.14 g/L to 7 g/L, respectively.
聚羟基脂肪酸酯(PHA)是一种具有许多潜在生物医学应用的可生物降解材料,包括医疗植入物和药物递送。本研究开发了一种用于筛选生产菌株的系统,以优化台湾贪铜菌184、185、186、187、204、208、209以及食油假单胞菌ATCC 29347中的PHA生产。在本研究中,苏丹黑B染色、红外(IR)和气相色谱(GC)分析表明,用于PHA合成的最佳菌株是台湾贪铜菌184,其可获得聚羟基丁酸酯(PHB)。在pH为7.0、30℃和搅拌速度为200 rpm的条件下培养台湾贪铜菌184,获得了10%的PHB含量和0.14 g/L的PHB产量。用于分析台湾贪铜菌184生产PHB的碳源和氮源类型分别为葡萄糖酸和NH₄Cl。还确定了PHB生产的最佳碳/氮比。当为台湾贪铜菌184选择8/1的碳/氮比时,本研究表明PHB含量为58.81%,PHB产量为2.44 g/L。两阶段发酵策略显著提高了PHB含量和PHB产量。在营养限制条件下的两阶段发酵策略下,台湾贪铜菌184的PHB含量为72%,PHB浓度为7 g/L。最后,实验结果证实,优化用于培养本地台湾贪铜菌184菌株的生长培养基和发酵条件,分别将PHB含量从10%大幅提高到72%,将PHB产量从0.14 g/L提高到7 g/L。