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蓝藻聚羟基丁酸酯(PHB):筛选、优化与表征

Cyanobacterial Polyhydroxybutyrate (PHB): Screening, Optimization and Characterization.

作者信息

Ansari Sabbir, Fatma Tasneem

机构信息

Cyanobacterial Biotechnology Laboratory, Department of Biosciences, Jamia Millia Islamia (Central University), New Delhi, India.

出版信息

PLoS One. 2016 Jun 30;11(6):e0158168. doi: 10.1371/journal.pone.0158168. eCollection 2016.

Abstract

In modern life petroleum-based plastic has become indispensable due to its frequent use as an easily available and a low cost packaging and moulding material. However, its rapidly growing use is causing aquatic and terrestrial pollution. Under these circumstances, research and development for biodegradable plastic (bioplastics) is inevitable. Polyhydroxybutyrate (PHB), a type of microbial polyester that accumulates as a carbon/energy storage material in various microorganisms can be a good alternative. In this study, 23 cyanobacterial strains (15 heterocystous and 8 non-heterocystous) were screened for PHB production. The highest PHB (6.44% w/w of dry cells) was detected in Nostoc muscorum NCCU- 442 and the lowest in Spirulina platensis NCCU-S5 (0.51% w/w of dry cells), whereas no PHB was found in Cylindrospermum sp., Oscillatoria sp. and Plectonema sp. Presence of PHB granules in Nostoc muscorum NCCU- 442 was confirmed microscopically with Sudan black B and Nile red A staining. Pretreatment of biomass with methanol: acetone: water: dimethylformamide [40: 40: 18: 2 (MAD-I)] with 2 h magnetic bar stirring followed by 30 h continuous chloroform soxhlet extraction acted as optimal extraction conditions. Optimized physicochemical conditions viz. 7.5 pH, 30°C temperature, 10:14 h light:dark periods with 0.4% glucose (as additional carbon source), 1.0 gl-1 sodium chloride and phosphorus deficiency yielded 26.37% PHB on 7th day instead of 21st day. Using FTIR, 1H NMR and GC-MS, extracted polymer was identified as PHB. Thermal properties (melting temperature, decomposition temperatures etc.) of the extracted polymer were determined by TGA and DSC. Further, the polymer showed good tensile strength and young's modulus with a low extension to break ratio comparable to petrochemical plastic. Biodegradability potential tested as weight loss percentage showed efficient degradation (24.58%) of PHB within 60 days by mixed microbial culture in comparison to petrochemical plastic.

摘要

在现代生活中,石油基塑料因其常被用作易于获取且成本低廉的包装和成型材料而变得不可或缺。然而,其使用量的迅速增长正导致水生和陆地污染。在这种情况下,研发可生物降解塑料(生物塑料)是不可避免的。聚羟基丁酸酯(PHB)是一种微生物聚酯,作为碳/能量储存物质在各种微生物中积累,它可能是一个很好的替代品。在本研究中,对23株蓝藻菌株(15株有异形胞的和8株无异形胞的)进行了PHB生产筛选。在念珠藻NCCU - 442中检测到最高的PHB含量(占干细胞重量的6.44%),在钝顶螺旋藻NCCU - S5中最低(占干细胞重量的0.51%),而在柱孢藻属、颤藻属和席藻属中未发现PHB。通过苏丹黑B和尼罗红A染色在显微镜下证实了念珠藻NCCU - 442中存在PHB颗粒。以甲醇:丙酮:水:二甲基甲酰胺[40:40:18:2(MAD - I)]对生物质进行预处理,磁力搅拌2小时,然后连续氯仿索氏提取30小时,这是最佳提取条件。优化的理化条件,即pH值7.5、温度30°C、光照:黑暗周期为10:14小时、添加0.4%葡萄糖(作为额外碳源)、1.0 g/L氯化钠以及缺磷,在第7天而非第21天产生了26.37%的PHB。使用傅里叶变换红外光谱(FTIR)、核磁共振氢谱(1H NMR)和气相色谱 - 质谱联用(GC - MS),将提取的聚合物鉴定为PHB。通过热重分析(TGA)和差示扫描量热法(DSC)测定了提取聚合物的热性能(熔点、分解温度等)。此外,该聚合物显示出良好的拉伸强度和杨氏模量,断裂伸长率低,与石化塑料相当。以失重百分比测试的生物降解潜力表明,与石化塑料相比,混合微生物培养在60天内可有效降解PHB(24.58%)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6fb/4928839/5dd15525d3bc/pone.0158168.g001.jpg

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