發(fā)布時(shí)間：2018-02-05 22:45

  本文關(guān)鍵詞： 側(cè)孢短芽孢桿菌 天然質(zhì)粒 異源表達(dá) 抗菌肽 抑菌效價(jià)　出處：《食品科學(xué)》2017年08期 　論文類型：期刊論文

【摘要】：側(cè)孢短芽孢桿菌抗菌肽具有高效、廣譜、穩(wěn)定的抗菌活性,該抗菌活性與其內(nèi)源質(zhì)粒密切相關(guān)。為研究側(cè)孢短芽孢桿菌含抗菌肽基因質(zhì)粒異源表達(dá)的可行性,本研究中將側(cè)孢短芽孢桿菌S62~-9的天然質(zhì)粒電轉(zhuǎn)枯草芽孢桿菌Wb800受體細(xì)胞,篩選陽(yáng)性轉(zhuǎn)化子并對(duì)其表達(dá)產(chǎn)物性質(zhì)進(jìn)行研究。得到了產(chǎn)生抗菌活性產(chǎn)物菌株Z4,發(fā)酵6 h后開(kāi)始產(chǎn)生抗菌物質(zhì),12~20 h產(chǎn)量達(dá)到最大,達(dá)620.96 AU/m L。Tricine~-十二烷基硫酸鈉~-聚丙烯酰胺凝膠電泳看到S62~-9質(zhì)粒在枯草芽孢桿菌中得到了成功的表達(dá),反相高效液相色譜分析Z4表達(dá)產(chǎn)物含有與側(cè)孢短芽孢桿菌抗菌肽相同物質(zhì)。該物質(zhì)抑菌譜廣,穩(wěn)定性好:對(duì)于G~+、G~-和真菌均有抑菌活性,而且對(duì)于G~+菌株的抑菌效果優(yōu)于G~-菌株和真菌,121℃處理20 min后的效價(jià)保留率約為90%,100℃處理60 min后仍有81%的活性保留;p H 2~12的范圍內(nèi)效價(jià)保留率基本不發(fā)生變化;對(duì)蛋白酶較為敏感,胰蛋白酶和蛋白酶K處理后抗菌肽的效價(jià)保留率為60%左右,其他3種蛋白酶處理后也有一定程度的降低。本研究獲得了抗菌肽的異源表達(dá)菌株Z4,為進(jìn)一步工業(yè)生產(chǎn)抗菌肽提供支持,而且為該抗菌肽遺傳基因定位于質(zhì)粒上提供了證明。
[Abstract]:The antimicrobial peptides of Bacillus parthenosporium have high efficiency, broad spectrum and stable antibacterial activity, which is closely related to its endogenous plasmids. This study is intended to study the feasibility of heterologous expression of antimicrobial peptide gene plasmids in Bacillus parthenosporium. In this study, the natural plasmids of Bacillus cereus S62C9 were electrotransferred into Wb800 receptor cells of Bacillus subtilis. The positive transformants were screened and the properties of the expressed products were studied. Strain Z4 was obtained and the production of the antibacterial substance began to reach the maximum at 20 h after fermentation for 6 h. As far as 620.96 AU/m L. Tricineto- 12 sodium alkyl sulfate and polyacrylamide gel electrophoresis, we can see that S62- 9 plasmid was successfully expressed in Bacillus subtilis (Bacillus subtilis). RP-HPLC analysis showed that the expressed product of Z4 contained the same substance as the antimicrobial peptide of Bacillus cereus. It had wide antibacterial spectrum and good stability. Moreover, the bacteriostasis effect of G- strain was better than that of G- strain and fungi treated at 121 鈩，

本文編號(hào)：1492958