食品科学 ›› 2020, Vol. 41 ›› Issue (20): 112-118.doi: 10.7506/spkx1002-6630-20190826-278

• 生物工程 • 上一篇    下一篇

戊糖乳杆菌LS1细菌素的分离纯化及性质鉴定

李志如,韩建春,刘容旭,刘丹怡,梁君锋   

  1. (1.东北农业大学食品学院,黑龙江 哈尔滨 150030;2.黑龙江省绿色食品科学研究院,黑龙江 哈尔滨 150028)
  • 出版日期:2020-10-25 发布日期:2020-10-23
  • 基金资助:
    中央引导地方科技项目(YDZX20172300004490)

Purification and Characterization of Bacteriocin Produced by Lactobacillus pentosus LS1

LI Zhiru, HAN Jianchun, LIU Rongxu, LIU Danyi, LIANG Junfeng   

  1. (1. College of Food Science, Northeast Agricultural University, Harbin 150030, China;2. Heilongjiang Green Food Research Institute, Harbin 150028, China)
  • Online:2020-10-25 Published:2020-10-23

摘要: 由酸菜水分离出的菌株LS1经16S rRNA鉴定为戊糖乳杆菌(Lactobacillus pentosus)。其细菌素首先通过硫酸铵沉淀取上清液经Sephadex G-10柱色谱脱盐并纯化,然后SP Sepharose Fast Flow中度纯化,最后Superdex 30 Increase精细纯化。纯化后比活力为1 238.1 AU/mg,是原始细菌素活性的110.5 倍。根据液相色谱-串联质谱仪测定细菌素分子质量为1 123.8 Da,氨基酸序列为ACDFCFCGMK。戊糖乳杆菌素Pentocin LS1显示出对部分革兰氏阴性和阳性食品腐败菌的抑菌活性。此外,100 ℃加热15 min仍保留80.2%的抑菌活性,在pH 2.0~8.0范围内具有较强活性,对蛋白水解酶敏感,对淀粉酶、脂肪酶及过氧化氢酶不敏感,部分化学试剂也对其活性无明显影响。结果表明,戊糖乳杆菌素LS1在食品防腐方面具有潜在应用价值。

关键词: 戊糖乳杆菌;纯化;细菌素;特性

Abstract: Bacteriocin-producing strain LS1 was isolated from Chinese sauerkraut juice and it was identified as Lactobacillus pentosus by 16S rRNA gene sequencing. The bacteriocin was separated from the fermentation supernatant of this strain by ammonium sulfate precipitation, desalted and purified by sequential column chromatography on Sephadex G-10, SP Sepharose Fast Flow, and Superdex 30 Increase. The specific activity of the purified bacteriocin was 1 238.1 AU/mg, which was 110.5 times higher than before the purification. The molecular mass of the bacteriocin was 1 123.8 Da and its N-terminal amino acid sequence was ACDFCFCGMK as determined by liquid chromatography-ion trap tandem mass spectrometry (LC-MS/MS). Pentocin LS1 showed a bacteriostatic activity against some Gram-negative and positive food spoilage bacteria. In addition, pentocin LS1 still retained 80.2% of its original antibacterial activity after being heated at 100 ℃ for 15 min, had strong activity in the range of pH 2.0-8.0, and was sensitive to proteolytic enzymes but insensitive to amylase, lipase catalase, and some chemical reagents. The results showed that pentocin LS1 can be potentially applied in food preservation.

Key words: Lactobacillus pentosus; purification; bacteriocin; characteristics

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