Saito Kyo, Reddy M Venkateswar, Sarkar Omprakash, Kumar A Naresh, Choi DuBok, Chang Young-Cheol
Course of Chemical and Biological Engineering, Division of Sustainable and Environmental Engineering, Muroran Institute of Technology, 27-1 Mizumoto, Muroran 050-8585, Japan.
Department of Civil and Environmental Engineering, Colorado State University, Fort Collins, CO 80523, USA.
Bioengineering (Basel). 2023 May 20;10(5):618. doi: 10.3390/bioengineering10050618.
With the growing interest in bioplastics, there is an urgent need to develop rapid analysis methods linked to production technology development. This study focused on the production of a commercially non-available homopolymer, poly(3-hydroxyvalerate) (P(3HV)), and a commercially available copolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB--3HV)), through fermentation using two different bacterial strains. The bacteria and sp. CYR1 were used to produce P(3HV) and P(3HB--3HV), respectively. The bacterium sp. CYR1 produced 415 mg/L of P(3HB--3HV) when incubated with acetic acid and valeric acid as the carbon sources, whereas the bacterium produced 0.198 g of P(3HV)/g dry biomass when incubated with sodium valerate as the carbon source. Additionally, we developed a fast, simple, and inexpensive method to quantify P(3HV) and P(3HB--3HV) using high-performance liquid chromatography (HPLC). As the alkaline decomposition of P(3HB--3HV) releases 2-butenoic acid (2BE) and 2-pentenoic acid (2PE), we were able to determine the concentration using HPLC. Moreover, calibration curves were prepared using standard 2BE and 2PE, along with sample 2BE and 2PE produced by the alkaline decomposition of poly(3-hydroxybutyrate) and P(3HV), respectively. Finally, the HPLC results obtained by our new method were compared using gas chromatography (GC) analysis.
随着对生物塑料的兴趣日益浓厚，迫切需要开发与生产技术发展相关的快速分析方法。本研究聚焦于通过使用两种不同的细菌菌株进行发酵来生产一种商业上不可用的均聚物聚（3-羟基戊酸酯）（P(3HV)）和一种商业上可用的共聚物聚（3-羟基丁酸酯- co - 3-羟基戊酸酯）（P(3HB-3HV)）。分别使用细菌 和sp. CYR1来生产P(3HV)和P(3HB-3HV)。当以乙酸和戊酸作为碳源进行培养时，细菌sp. CYR1产生了415 mg/L的P(3HB-3HV)，而当以戊酸钠作为碳源进行培养时，细菌 产生了0.198 g P(3HV)/g干生物量。此外，我们开发了一种快速、简单且廉价的方法，使用高效液相色谱（HPLC）对P(3HV)和P(3HB-3HV)进行定量。由于P(3HB-3HV)的碱性分解会释放2-丁烯酸（2BE）和2-戊烯酸（2PE），我们能够使用HPLC测定其浓度。此外，使用标准的2BE和2PE以及分别由聚（3-羟基丁酸酯）和P(3HV)的碱性分解产生的样品2BE和2PE制备了校准曲线。最后，将我们新方法获得的HPLC结果与气相色谱（GC）分析结果进行了比较。
