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采用单因素法(OFAT)从阿氏假单胞菌(PS1)分离株中生产和表征糖脂生物表面活性剂,并合理利用氨浸预处理木质纤维素残渣。

Production and characterization of glycolipid biosurfactant from Achromobacter sp. (PS1) isolate using one-factor-at-a-time (OFAT) approach with feasible utilization of ammonia-soaked lignocellulosic pretreated residues.

机构信息

Amity Institute of Biotechnology, Amity University, Sector- 125, Noida, UP, 201313, India.

School of Biological Science, University of Portsmouth, Portsmouth, PO1 2DY, UK.

出版信息

Bioprocess Biosyst Eng. 2019 Aug;42(8):1301-1315. doi: 10.1007/s00449-019-02128-3. Epub 2019 Apr 26.

Abstract

With the ever growing increase in the demands of biosurfactants, the present study was focused in developing a set of parameters influencing biosurfactant production using one-factor-at-a-time (OFAT) approach in chemically defined medium from an indigenous isolate of Achromobacter sp. (PS1). Subsequently, the feasibility of biosurfactant production was examined using influential OFAT parameters in same medium, replacing only carbon source with lignocellulosic hydrolyzed sugars. These sugars were obtained from ammonia (15% v/v) soaking pretreatment of lignocellulosic residues (7.5% solid loading at 70 °C for 72 h) with subsequent saccharification using lignocellulolytic enzymes. OFAT influential parameters observed were dextrose (3-4% w/v); C/N ratio 8.3 using sodium nitrate and beef extract; 2 × 10 grams equivalents Fe; 1500 mM PO in minimal salt medium (MSM) at pH 7.0, 120 rpm, 30 °C resulting in 4.13 ± 0.12 g/L rhamnolipid in 192 h with 30.42 mN/m surface tension and 136 mg/L critical micelle concentration (CMC). Biosurfactant was characterized using tandem-MS and NMR as rhamnolipid with six-congeners, Rha-C-C and Rha-Rha-C-C being the most abundant. Rhamnolipid showed broad range stability at temperatures (30-121 °C), pH (6-12), and salinity (0.5-5% w/v) of NaCl. In Rice-straw (RS) hydrolysate, maximum glucan (73.10%) and xylan (91.13%) were obtained and the RS-hydrolysate medium with a total of 4.55% (w/v) sugars under optimum OFAT parameters (other than dextrose) showed at par production of 3.55 ± 0.06 g/L of rhamnolipid in 192 h with Y (biosurfactant yield per gram of sugar consumed) of 0.08 g/g and Y (biosurfactant yield per gram of cell biomass) of 0.68 g/g.

摘要

随着生物表面活性剂需求的不断增长,本研究采用单因素分析法(OFAT),在一种从 Achromobacter sp.(PS1)分离得到的纯培养物的化学成分确定的培养基中,研究了影响生物表面活性剂生产的一系列参数。随后,在相同的培养基中,仅用木质纤维素水解糖替代碳源,考察了生物表面活性剂生产的可行性。这些糖是从木质纤维素残渣的氨(15%v/v)浸泡预处理(70°C 下 7.5%固含量 72 小时)中获得的,然后用木质纤维素酶进行糖化。观察到的 OFAT 影响参数为葡萄糖(3-4%w/v);C/N 比为 8.3,使用硝酸钠和牛肉提取物;2×10 克当量的 Fe;在最小盐培养基(MSM)中添加 1500 mM PO,pH7.0、120 rpm、30°C,在 192 小时内产生 4.13±0.12g/L 鼠李糖脂,表面张力为 30.42 mN/m,临界胶束浓度(CMC)为 136mg/L。生物表面活性剂采用串联-MS 和 NMR 进行了表征,结果表明其为六同系物鼠李糖脂,其中 Rha-C-C 和 Rha-Rha-C-C 最为丰富。鼠李糖脂在温度(30-121°C)、pH(6-12)和盐度(0.5-5%w/v)的 NaCl 范围内具有广泛的稳定性。在水稻秸秆(RS)水解物中,获得了最高的葡聚糖(73.10%)和木聚糖(91.13%),在最佳 OFAT 参数(除葡萄糖外)下,RS 水解物培养基中的总糖含量为 4.55%(w/v),在 192 小时内产生了 3.55±0.06g/L 的鼠李糖脂,糖消耗的生物表面活性剂产率(Y)为 0.08g/g,细胞生物质的生物表面活性剂产率(Y)为 0.68g/g。

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