以发酵上清液表面活性的排油活性、乳化能力、表面张力为初筛参数,srfA基因为复筛指标,定向筛选产表面活性剂surfactin的菌株。通过薄层层析(thin layer chromatography,TLC)定性分析和超高效液相色谱-电喷雾四级杆飞行时间质谱(ultra performance liquid chromatography/electrospray ionization quadrupole time-of-flight high-resolution/mass spectrometry,UPLC-ESI/Q-TOF/MS)定量分析,筛选获得1株可利用啤酒糟高产surfactin的菌株41-3,鉴定为枯草芽孢杆菌(Bacillus subtilis),采用液质分析所产surfactin同系物组分。B. subtilis 41-3在MSM培养基中发酵,surfactin产量为541.32 mg/L,同系物以脂肪酸链为C14和C13的surfactin为主,两者含量约占75%。以啤酒糟酶解液作为碳源,发酵生产surfactin,产量提高1.38倍,达到746.35 mg/L。且surfactin的合成向长链脂肪酸surfactin迁移,表明其具有更强的生物表面活性。该研究结果为surfactin菌株筛选提供新的思路,为发酵生产surfactin提供了新的原料来源,为啤酒糟综合利用提供新的途径,具有应用前景。
The surfactin producing strains were screened directionally using oil repellency, emulsifying ability and surface tension of the fermentation supernatant surfactant as preliminary screening parameters, and srfA gene for rescreening. A surfactin high-yield strain 41-3 utilizing brewer's spent grain (BSG) was screened by thin layer chromatography (TLC) qualitative analysis and ultra-performance liquid chromatography/electrospray ionization quadrupole time-of-flight high-resolution/mass spectrometry (UPLC-ESI/Q-TOF MS) quantitative analysis. The strain was identified as Bacillus subtilis, and different congeners of surfactin were analyzed. The surfactin yield of B. subtilis 41-3 was 541.32 mg/L in MSM medium, and C14 and C13 fatty-acid-chain surfactins were the two most abundant homologues, which accounted for 75% of total surfactin. When BSG was used as fermentation substrate, the yield of surfactin increased 1.38 times to 746.35 mg/L, and the surfactin synthesis shifted to long fatty-acid-chain surfactins, which indicated better biosurface activity. This study provides new ideas for surfactin-producing strain screening and new raw materials for surfactin production, as well as revealing a new way for comprehensive utilization of BSG.
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