Optimization of Physical Parameters Involved in Cell Lysis of Bacillus Sp. to Recover Bioplastic Polyhydroxyalkanoates

S. G. Bhat, D. B. Kodavoor, S. Raja, T. Varadavenkatesan, D. M. Somashekara
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Abstract

Abstract The increased global population has concurrently increased waste disposal, whose majority is conventional plastic. In this study, polyhydroxyalkanoates (PHA), an alternative biopolymer to conventional plastics, were extracted from bacteria Bacillus sp ., using response surface methodology (RSM), a statistical approach. To design, optimize and study the relationship between the parameters (glass beads weight, incubation time, water volume, incubation temperature, and shaker speed) Box-Behnken Design of response surface methodology was applied in Design Expert 10.0 software package. The solvent method is known in PHA extraction; however, this approach is environmentally hazardous on a large scale. The current study used a physical extraction method using glass beads for bacterial cell lysis. As a characterization, FTIR, 1HNMR, and DSC confirmed the recovered polymer as Polyhydroxy butyrate (PHB). 31.53% (w/v) of PHB was recovered for 1 g/L biomass. PHB is known to be widely applied in various fields, specifically in medical applications. Genetically modified isolate, low-cost substrate, and recovery without solvent assure a cost-effective and increased PHA production. Glass beads can be reused in extraction, reducing overall production cost. Therefore, this work used a reduced amount of chemicals during extraction to recover the PHB. Thus, sustainability assures a better scope for the future promotion of PHA production in academia and industries. Highlights Sustainable PHA extraction alternatives, reducing environmental impact, were studied. Glass beads are used to lyse the cell to release the PHA from the bacteria Bacillus sp. Box-Behnken design optimized extraction parameters for maximum recovery of PHA. The cellular lysis approach yields higher recovery than solvent-method recovery and is eco-friendly. Graphical abstract
芽孢杆菌裂解细胞回收生物塑料聚羟基烷酸酯物理参数的优化
随着全球人口的增长,垃圾处理也随之增加,其中大部分是传统塑料。本研究采用响应面法(RSM)从芽孢杆菌(Bacillus sp .)中提取了一种可替代传统塑料的生物聚合物聚羟基烷酸酯(PHA)。为了设计、优化和研究玻璃微珠重量、孵育时间、水量、孵育温度和激振器转速等参数之间的关系,在design Expert 10.0软件包中应用响应面法的Box-Behnken design。溶剂法是已知的PHA提取法;然而,这种方法在很大程度上对环境有害。目前的研究采用物理提取法,利用玻璃微珠进行细菌细胞裂解。通过FTIR、1HNMR和DSC表征,证实该聚合物为聚羟基丁酸酯(Polyhydroxy butyrate, PHB)。1 g/L生物量中PHB的回收率为31.53% (w/v)。PHB被广泛应用于各个领域,特别是在医学应用中。转基因分离物、低成本底物和无溶剂回收确保了成本效益和PHA产量的增加。玻璃珠可以在萃取中重复使用,降低了整体生产成本。因此,这项工作在提取过程中使用了少量的化学物质来回收PHB。因此,可持续性保证了未来在学术界和工业界促进PHA生产的更好范围。重点研究了可持续的PHA提取方法,减少了对环境的影响。利用玻璃微珠裂解细胞,释放细菌芽孢杆菌中的PHA。Box-Behnken设计优化提取工艺,最大限度地回收PHA。细胞裂解法比溶剂法回收率高,而且环保。图形抽象
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