摘要
目的:对戊糖乳杆菌31-1的群体感应信号肽进行提取、纯化和鉴定。方法:经硫酸铵盐析、超滤、疏水层析、凝胶层析和RP-HPLC步骤纯化戊糖乳杆菌31-1的群体感应信号肽。用MALDI-TOF质谱测定纯化后的小肽AIP的分子质量,并用EDMAN降解法对其N端氨基酸测序。用组氨酸蛋白激酶抑制剂——氯氰碘柳胺验证其群体感应信号肽分子功能。结果:纯化的信号肽AIP的比活力达160 000 IU/m L,纯度提高了800倍。该小肽的分子质量为2 985.16 u。其N端前15个氨基酸序列为NH2-KSSAYSLQMGATAIK。氯氰碘柳胺所造成的戊糖乳杆菌31-1细菌素合成功能的缺失,不能通过添加小肽AIP而得以恢复,证实了小肽AIP的群体感应信号肽分子功能。结论:群体感应信号肽AIP的获得为戊糖乳杆菌31-1细菌素的发酵调控奠定了基础。
Objective: To purify and identify quorum sensing signal peptide of Lactobacillus pentosus 31-1. Methods: The active principle was purified to homogeneity by ammonium sulfate precipitation, ultrafilteration, hydrophobic interac- tion chromatography, gel filtration chromatography and reverse phase high-performance liquid chromatography (RP-HPLC). MALDI-TOF mass spectrometer was used for determination molecular weight of the purified peptide AIP, and its N-ter- minal sequences was analyzed by Edman degradation. Closantel, a histidine protein kidnase inhibitor was used to verify the role of the purified peptide. Results: The purification resulted in an 800-fold increase in specific activity with a yield of 160 000 IU/mL of the induction activity. The mass of the purified peptide AIP was 2 985.16 u, its partial N-ter- minal sequence was: NH2-KSSAYSLQMGATAIK. Bacteriocin production inhibited by closantel could not be restored by peptide AlP, which further confirmed its quorum sensing signal molecule role. Conclusion: The obtaining of the quorum sensing peptide AIP laid the fundation for the regulation of bacteriocin fermentation in L. pentosus 31-1.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2015年第3期201-206,共6页
Journal of Chinese Institute Of Food Science and Technology
基金
国家自然科学基金项目(31071591
31471707)
河北省高等学校科学技术研究项目(ZD20131091)
关键词
细菌素
戊糖乳杆菌
纯化
特性
bacteriocin
Lactobacillus pentosus
purification
characterization