Israni Neetu, Thapa Surabhi, Shivakumar Srividya
Department of Microbiology, School of Sciences, Jain University, Bangalore 560011, India.
J Genet Eng Biotechnol. 2018 Dec;16(2):265-271. doi: 10.1016/j.jgeb.2018.07.004. Epub 2018 Jul 21.
The applicability of sp. cell lytic enzymes for devising a simple and competent biological polyhydroxyalkanoate (PHA) recovery approach from cells was investigated. strain Ti3 produced 50% (w/w) PHA using glucose as carbon source. The intracellular PHA was recovered employing a non-PHA accumulating actinomycetes (Tia1) identified as , having potent lytic activity against living and heat inactivated . Interestingly, maximum biomass (2.53 ± 0.6 g/L by 24 h) of the lytic actinomycete was obtained in PHA production medium itself thus circumventing the prior actinomycete acclimatization just by co-inoculation with as an inducer. Maximum lytic activity was observed at pH 6.0, 40 °C, 220 mg of biomass and 33.3 mL of concentrated culture filtrate in a 100 mL reaction mixture. Preliminary biochemical investigations confirmed the proteolytic and caseinolytic nature of the lytic enzyme. PHA yield of 0.55 g/g by co-inoculation extraction approach was comparable with the conventional sodium hypochlorite based extraction method. Interestingly, also demonstrated a broad spectrum lytic potential against varied Gram-negative and Gram-positive PHA producers highlighting the extensive applicability of this biolytic PHA recovery approach. The lytic enzyme retained almost 100% relative activity on storage at -20 °C upto two months. H Nuclear magnetic resonance analysis of the extracted polymer confirmed it as a homopolymer composed of 3-hydroxybutyrate monomeric units. This is the first report on sp. based biological and eco-friendly, intracellular PHA recovery from spp.
研究了特定细胞裂解酶在设计一种从细胞中简单且有效的生物聚羟基脂肪酸酯(PHA)回收方法中的适用性。菌株Ti3以葡萄糖作为碳源生产了50%(w/w)的PHA。利用一种被鉴定为对活细胞和热灭活细胞具有强大裂解活性的非PHA积累放线菌(Tia1)回收细胞内的PHA。有趣的是,在PHA生产培养基本身中就获得了裂解放线菌的最大生物量(24小时时为2.53±0.6 g/L),因此仅通过与作为诱导剂的共同接种就避免了先前的放线菌驯化。在100 mL反应混合物中,在pH 6.0、40°C、220 mg生物量和33.3 mL浓缩培养滤液的条件下观察到最大裂解活性。初步生化研究证实了裂解酶的蛋白水解和酪蛋白水解性质。通过共同接种提取方法获得的PHA产量为0.55 g/g,与传统的基于次氯酸钠的提取方法相当。有趣的是,还对多种革兰氏阴性和革兰氏阳性PHA生产者表现出广谱裂解潜力,突出了这种生物裂解PHA回收方法的广泛适用性。裂解酶在-20°C下储存长达两个月时保留了几乎100%的相对活性。对提取聚合物的核磁共振分析证实它是一种由3-羟基丁酸单体单元组成的均聚物。这是关于基于特定菌株从特定物种中进行生物且环保的细胞内PHA回收的首次报道。