Narayanan Mathiyazhagan, Kandasamy Sabariswaran, Kumarasamy Suresh, Gnanavel Keerthana, Ranganathan Muthusamy, Kandasamy Gajendiran
PG and Research Centre in Biotechnology, MGR College, Adhiyamaan Educational Research Institute, Hosur, Krishnagiri, Tamilnadu, India.
Institute for Energy Research, Jiangsu University, Zhenjiang, China.
Heliyon. 2020 Oct 30;6(10):e05381. doi: 10.1016/j.heliyon.2020.e05381. eCollection 2020 Oct.
The prime aim of this study was to enumerate predominant bacteria from polluted lake soil samples, which possess polyhydroxybutyrate (PHB) fabricating potential and identify the suitable growth conditions and nutritional factors for PHB fabrication. From several numbers of bacterial cultures, one culture has the competence to yield PHB, and it was endorsed through Sudan Black B stain, Nile red staining, SEM analysis, and growth in PHB selective media. Under the microscopic observation, the fluorescent cells and polymeric granules were observed in the fluorescent microscope and SEM, respectively. This PHB fabricating isolate was recognized as NDRMN001 through 16S rRNA partial sequence analysis. The structural characteristics of PHB produced by NDRMN001 were studied through FT-IR, H NMR, and C NMR analysis. The peak observed at 1759.27 cm on FT-IR analysis is corresponding to the signal band of PHB. In H NMR peaks were noticed at 1.67, 2.37 to 2.71, and 3.38 to 7.68 which corresponding to -CH3, -CH2, and -CH protons of PHB. About 4 notable peaks were noticed in C NMR analysis at 19.62, 68.27, 40.68, and 169.11 ppm which appeared close to the carboxyl group of PHB. About 10% of inoculum, pH 7.5, 2 g L of yeast extract, 20 g L of rice bran, 35 °C, and 2 days of incubation were recognized as optimal growth conditions for NDRMN001 to produce PHB. The identified NDRMN001 has the potential to yield 91.48% of PHB as 33.19 g L of PHB from 36.26 g L of culture biomass. The complete results conclude that the NDRMN001 screened from polluted lake soil has the competence to produce fine quality and quantity of PHB in a short duration of fabrication process under favorable conditions with the utilization of cheap nutritional factors.
本研究的主要目的是从受污染的湖泊土壤样本中筛选出具有聚羟基丁酸酯(PHB)合成潜力的优势细菌,并确定适合PHB合成的生长条件和营养因素。从多个细菌培养物中,有一种培养物具有产生PHB的能力,并通过苏丹黑B染色、尼罗红染色、扫描电子显微镜(SEM)分析以及在PHB选择性培养基中的生长情况得到了证实。在显微镜观察下,分别在荧光显微镜和SEM中观察到了荧光细胞和聚合物颗粒。通过16S rRNA部分序列分析,这种PHB合成菌株被鉴定为NDRMN001。通过傅里叶变换红外光谱(FT-IR)、氢核磁共振(H NMR)和碳核磁共振(C NMR)分析研究了NDRMN001产生的PHB的结构特征。FT-IR分析中在1759.27 cm处观察到的峰对应于PHB的信号带。在H NMR中,在1.67、2.37至2.71以及3.38至7.68处观察到峰,分别对应于PHB的-CH3、-CH2和-CH质子。在C NMR分析中,在19.62、68.27、40.68和169.11 ppm处观察到约4个显著峰,这些峰靠近PHB的羧基。约10%的接种量、pH 7.5、2 g/L的酵母提取物、20 g/L的米糠、35°C以及2天的培养时间被认为是NDRMN001产生PHB的最佳生长条件。鉴定出的NDRMN001能够从36.26 g/L的培养生物量中产生33.19 g/L的PHB,占PHB的91.48%。完整的结果表明,从受污染的湖泊土壤中筛选出的NDRMN001有能力在有利条件下,利用廉价的营养因素,在短时间的合成过程中生产出高质量和高产量的PHB。