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一株高效表面活性剂产生菌的筛选及其活性产物的结构与性质

肖兵1,2,杨德玉1,2,史荣久2**,张颖2,张阳1,郝金生3,国桥3,劲毅3   

  1. (1沈阳师范大学化学与生命科学学院, 沈阳 110034; 2中国科学院沈阳应用生态研究所, 中国科学院污染生态与环境工程 重点实验室, 沈阳 110016; 3大庆油田有限责任公司第二采油厂中心化验室, 黑龙江省大庆 163414)
  • 出版日期:2013-03-10 发布日期:2013-03-10

Screening of an efficient biosurfactant-producing bacterial strain and the structure and property of its surface-active metabolic product.

XIAO Bing1,2, YANG De-yu1,2, SHI Rong-jiu2**, ZHANG Ying2, ZHANG Yang1, HAO Jin-sheng3, LI Guo-qiao3, ZHAO Jin-yi3   

  1. (1College of Chemistry and Life Sciences, Shenyang Normal University, Shenyang 110034, China; 2Key Laboratory of Terrestrial Ecological Processes, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China; 3Center Laboratory of No. 2 Oil Production Plant, Daqing Oilfield Company Ltd., Daqing 163414, Heilongjiang, China)
  • Online:2013-03-10 Published:2013-03-10

摘要: 采用血平板法结合表面张力法从大庆油田高台子低渗透油藏采出水中筛选出一株高效表面活性剂产生菌,编号为DY-9。16S rDNA序列同源性比对结果显示,DY-9与地衣芽孢杆菌(Bacillus licheniformis)的序列相似性达99%。在45 ℃、180 r·min-1振荡培养条件下,DY-9在20 g·L-1糖蜜为碳源的培养基中生长良好,24 h内将发酵液的表面张力从初始的69.3 mN·m-1降低至26.5 mN·m-1。48 h发酵液中表面活性剂粗品的产量达0.48 g·L-1,其临界胶束浓度为28 mg·L-1。菌株产生的表面活性剂在高温(120 ℃,1 h)、高盐(12% NaCl)及较宽的pH(2~12)条件下保持稳定,溶液的表面张力低于30 mN·m-1。经傅里叶变换红外光谱(IF-TR)和高效液相色谱-质谱联用(HPLC-MS)分析,菌株DY-9的表面活性产物为环状脂肽类表面活性剂。

关键词: 烤烟, 潜香型物质, 稳定性, 基因型与环境互作, AMMI模型

Abstract: By adopting blood-agar procedure combined with surface tension screening strategy, an efficient surfactant-producing bacterial strain DY-9 was successfully isolated from the produced water from low-permeability Gaotaizi oil reservoir, Daqing Oilfield of Northeast China. The comparison with the homoeologous 16S rDNA sequences deposited in the GenBank showed that the DY9 had a 99% sequence similarity with Bacillus licheniformis. When cultured on a medium with 20 g·L-1 of molasses as the sole carbon source at 180 r·min-1 and 45 °C, the DY-9 could grow well, and decrease the surface tension of culture broth from initial 69.3 mN·m-1 to 26.5 mN·m-1 within 24 h. After cultured for 48 h, the maximum raw biosurfactant yield of the strain reached 0.48 g·L-1, and the critical micelle concentration was 28 mg·L-1. The biosurfactant produced by the DY9 could keep stable under the conditions of high temperature (120 °C) for 1 h, high salinity (12% NaCl), and wide pH (2-12), and the surface tension of the zymotic fluid within 48h was lower than 30 mN·m-1. The analysis with infrared spectroscopy (IF-TR) and high-pressure liquid chromatographymass spectrometry (HPLC-MS) revealed that the surface-active product of the DY-9 was a cyclic structure lipopeptide biosurfactant.

Key words: tobacco, latently fragrant substance, stability, genotype and environment interaction, AMMI model.