Dai Yu, Yuan Zhiguo, Jack Kevin, Keller Jurg
Advanced Wastewater Management Centre (AWMC), The University of Queensland, St. Lucia, Brisbane 4072, Australia.
J Biotechnol. 2007 May 1;129(3):489-97. doi: 10.1016/j.jbiotec.2007.01.036. Epub 2007 Feb 11.
One of the main limitations in bacterial polyhydroxyalkanoate (PHA) production with mixed cultures is the fact that primarily polyhydroxybutyrate (PHB) homopolymers are generated from acetate as the main carbon source, which is brittle and quite fragile. The incorporation of different 3-hydroxyalkanoate (HA) components into the polymers requires the addition of additional carbon sources, leading to extra costs and complexity. In this study, the production of poly(3-hydroxybutyrate (3HB)-co-3-hydroxyvalerate (3HV)-co-3-hydroxy-2-methylvalerate (3HMV)), with 7-35C-mol% of 3HV fractions from acetate as the only carbon source was achieved with the use of glycogen accumulating organisms (GAOs). An enriched GAO culture was obtained in a lab-scale reactor operated under alternating anaerobic and aerobic conditions with acetate fed at the beginning of the anaerobic period. The production of PHAs utilizing the enriched GAO culture was investigated under both aerobic and anaerobic conditions. A polymer content of 14-41% of dry cell weight was obtained. The PHA product accumulated by GAOs under anaerobic conditions contained a relatively constant proportion of non-3HB monomers (30+/-5C-mol%), irrespective of the amount of acetate assimilated. In contrast, under aerobic conditions, GAOs only produced 3HB monomers from acetate causing a gradually decreasing 3HV fraction during this aerobic feeding period. The PHAs were characterized by gel permeation chromatography (GPC) and differential scanning calorimetry (DSC). The data demonstrated that the copolymers possessed similar characteristics to those of commercially available poly(3HB-co-3HV) (PHBV) products. The PHAs produced under solely anaerobic conditions possessed lower melting points and crystallinity, higher molecular weights, and narrower molecular-weight distributions, compared to the aerobically produced polymers. This paper hence demonstrates the significant potential of GAOs to produce high quality polymers from a simple and cheap carbon source, contributing considerably to the growing research body on bacterial PHA production by mixed cultures.
利用混合培养物生产细菌聚羟基脂肪酸酯(PHA)的主要限制之一是,以乙酸盐作为主要碳源时,主要生成的是聚羟基丁酸酯(PHB)均聚物,而这种聚合物质地脆且相当易碎。将不同的3-羟基脂肪酸酯(HA)组分掺入聚合物中需要添加额外的碳源,这会导致额外成本和复杂性增加。在本研究中,使用糖原积累微生物(GAO)以乙酸盐作为唯一碳源实现了聚(3-羟基丁酸酯(3HB)-共-3-羟基戊酸酯(3HV)-共-3-羟基-2-甲基戊酸酯(3HMV))的生产,其中3HV组分的摩尔分数为7-35C-%。在实验室规模的反应器中,通过在厌氧和好氧交替条件下运行,并在厌氧期开始时加入乙酸盐,获得了富集的GAO培养物。在好氧和厌氧条件下研究了利用富集的GAO培养物生产PHA的情况。获得了占干细胞重量14-41%的聚合物含量。GAO在厌氧条件下积累的PHA产物中,非3HB单体的比例相对恒定(30±5C-%),与同化的乙酸盐量无关。相比之下,在好氧条件下,GAO仅从乙酸盐中产生3HB单体,导致在这个好氧进料期间3HV组分逐渐减少。通过凝胶渗透色谱法(GPC)和差示扫描量热法(DSC)对PHA进行了表征。数据表明,这些共聚物具有与市售聚(3HB-共-3HV)(PHBV)产品相似的特性。与好氧生产的聚合物相比,仅在厌氧条件下生产的PHA具有较低的熔点和结晶度、较高的分子量以及较窄的分子量分布。因此,本文证明了GAO从简单廉价的碳源生产高质量聚合物的巨大潜力,这对混合培养细菌生产PHA的不断增长的研究做出了重要贡献。