Chang Young-Cheol, Reddy Motakatla Venkateswar, Imura Kazuma, Onodera Rui, Kamada Natsumi, Sano Yuki
Course of Chemical and Biological Engineering, Division of Sustainable and Environmental Engineering, Muroran Institute of Technology, Hokkaido 050-8585, Japan.
Course of Biosystem, Department of Applied Sciences, Muroran Institute of Technology, Hokkaido 050-8585, Japan.
Bioengineering (Basel). 2021 Oct 24;8(11):157. doi: 10.3390/bioengineering8110157.
Cheese whey (CW) can be an excellent carbon source for polyhydroxyalkanoates (PHA)-producing bacteria. Most studies have used CW, which contains high amounts of lactose, however, there are no reports using raw CW, which has a relatively low amount of lactose. Therefore, in the present study, PHA production was evaluated in a two-stage process using the CW that contains low amounts of lactose. In first stage, the carbon source existing in CW was converted into acetic acid using the bacteria, C1, which was isolated from food waste. In the second stage, acetic acid produced in the first stage was converted into PHA using the bacteria, sp. CYR-1. Under the condition of without the pretreatment of CW, acetic acid produced from CW was diluted at different folds and used for the production of PHA. Strain CYR-1 incubated with 10-fold diluted CW containing 5.7 g/L of acetic acid showed the higher PHA production (240.6 mg/L), whereas strain CYR-1 incubated with four-fold diluted CW containing 12.3 g/L of acetic acid showed 126 mg/L of PHA. After removing the excess protein present in CW, PHA production was further enhanced by 3.26 times (411 mg/L) at a four-fold dilution containing 11.3 g/L of acetic acid. Based on Fourier transform infrared spectroscopy (FT-IR), and H and C nuclear magnetic resonance (NMR) analyses, it was confirmed that the PHA produced from the two-stage process is poly-β-hydroxybutyrate (PHB). All bands appearing in the FT-IR spectrum and the chemical shifts of NMR nearly matched with those of standard PHB. Based on these studies, we concluded that a two-stage process using C1 and sp. CYR-1 would be applicable for the production of PHB using CW containing a low amount of lactose.
奶酪乳清(CW)可以成为生产聚羟基脂肪酸酯(PHA)的细菌的优质碳源。大多数研究使用的是含有大量乳糖的CW,然而,尚无使用乳糖含量相对较低的未加工CW的相关报道。因此,在本研究中,采用含有少量乳糖的CW,通过两阶段工艺评估PHA的生产情况。在第一阶段,利用从食物垃圾中分离出的C1细菌将CW中存在的碳源转化为乙酸。在第二阶段,利用CYR-1菌将第一阶段产生的乙酸转化为PHA。在不对CW进行预处理的条件下,将CW产生的乙酸进行不同倍数的稀释,用于PHA的生产。用含有5.7 g/L乙酸的10倍稀释CW培养的CYR-1菌株显示出较高的PHA产量(240.6 mg/L),而用含有12.3 g/L乙酸的4倍稀释CW培养的CYR-1菌株显示出126 mg/L的PHA产量。去除CW中存在的过量蛋白质后,在含有11.3 g/L乙酸的4倍稀释条件下,PHA产量进一步提高了3.26倍(411 mg/L)。基于傅里叶变换红外光谱(FT-IR)以及氢和碳核磁共振(NMR)分析,证实两阶段工艺生产的PHA是聚-β-羟基丁酸酯(PHB)。FT-IR光谱中出现的所有谱带以及NMR的化学位移与标准PHB的几乎匹配。基于这些研究,我们得出结论,使用C1和CYR-1菌的两阶段工艺适用于利用含有少量乳糖的CW生产PHB。