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  本文選題：膽鹽水解酶 + 酶活力�。� 參考：《微生物學通報》2017年12期

【摘要】：【目的】隨著抗生素生長促進劑(AGPs)在動物飼料中逐步禁止使用,AGPs替代物的研究成為熱點。由于膽鹽水解酶(BSH)在脂類代謝中的關鍵作用,成為AGPs替代物研究的一個重要方向。在原核表達和純化的基礎上鑒定雞源和豬源乳桿菌BSH在酶學性質方面的差異性�！痉椒ā糠謩e對雞源膽鹽水解酶(BSHc)和豬源膽鹽水解酶(BSHp)基因進行原核表達和蛋白純化,通過測定對6種甘氨結合膽鹽和�；墙Y合膽鹽的水解效率獲得兩種酶的酶學動力學性質,進而測定了溫度、pH和金屬離子對酶活力的影響�！窘Y果】BSHc和BSHp對甘氨結合膽鹽的水解效率高于�；墙Y合膽鹽,BSHc對甘氨結合膽鹽的水解效率較BSHp稍高;BSHc和BSHp的最適酶解溫度分別為45°C和42°C;BSHc和BSHp的最適反應pH分別為6.0和5.4;含有Cu~(2+)、Fe~(3+)、Mn~(2+)和Zn~(2+)的金屬鹽對BSHc和BSHp的酶活力均具有不同程度的抑制作用,特別是Cu~(2+)和Fe~(3+)抑制作用比較強;含有Na~+、K~+、Mg~(2+)和Ca2+的金屬鹽對BSHc和BSHp酶活力的抑制作用相對較弱或無抑制作用,但KIO3對BSHc和BSHp酶活力具有強抑制作用,KI和CaCl_2對BSHp酶活力也具有較強的抑制作用�！窘Y論】原核表達和純化的BSHc和BSHp對甘氨結合膽鹽的水解效率高于�；墙Y合膽鹽,BSHc的最適酶解溫度和pH稍高于BSHp,Cu~(2+)、Fe~(3+)、Mn~(2+)和Zn~(2+)等金屬離子對BSHc和BSHp酶活力具有明顯抑制作用,試驗結果為鑒定BSH抑制物進而研制AGPs替代物奠定了基礎。
[Abstract]:[objective] as the antibiotic growth promoter AGPs has gradually banned the use of AGPs substitutes in animal feed, it has become a hot topic. Because of the key role of bile salt hydrolase BSHs in lipid metabolism, BSHs have become an important research direction in the research of AGPs substitutes. On the basis of prokaryotic expression and purification, the differences of enzymatic properties between chicken and porcine Lactobacillus BSH were identified. [methods] the prokaryotic expression and protein purification of chicken bile salt hydrolase BSHc and porcine bile salt hydrolase BSHppc were carried out, respectively. The enzymatic kinetic properties of the two enzymes were obtained by determining the hydrolysis efficiency of 6 kinds of glycine bound bile salts and taurosulfonated bile salts. Furthermore, the effects of temperature pH and metal ions on enzyme activity were determined. [results] the hydrolysis efficiency of BSHc and BSHp on glycosamino-bound bile salt was higher than that of tauro-bound bile salt BSHc on glycosamino-bound bile salt, which was slightly higher than that of BSHp and BSHp. The optimum pH for enzymatic hydrolysis at 45 擄C and 42 擄C for BSHc and BSHp were 6.0 and 5.4, respectively. Metal salts containing Cu~(2 and Zn~(2) inhibited the enzyme activities of BSHc and BSHp in different degrees. In particular, Cu~(2) and Fe~(3) had strong inhibitory effects on the enzyme activities of BSHc and BSHp, and the metal salts containing Na ~ + K ~ (2) and Ca2 had relatively weak or no inhibitory effects on the activities of BSHc and BSHp. But KIO3 can inhibit the activity of BSHc and BSHp. Ki and CaCl_2 have strong inhibition on BSHp activity. [conclusion] the hydrolytic efficiency of prokaryotic expression and purified BSHc and BSHp is higher than that of taurocholate. [conclusion] the hydrolysis efficiency of prokaryotic expression and purified BSHc and BSHp is higher than that of taurocholate. The optimum enzymatic hydrolysis temperature and pH of BSHc were slightly higher than those of BSHpCuO2) and Zn~(2), and the activities of BSHc and BSHp were inhibited obviously. The results laid a foundation for the identification of BSH inhibitors and the development of AGPs substitutes.
【作者單位】： 北京市農林科學院畜牧獸醫(yī)研究所畜禽疫病防控技術北京市重點實驗室;
【基金】：國家自然科學基金項目(No.31572527) 北京市農林科學院科技創(chuàng)新能力建設專項項目(No.KJCX20150703,KJCX20161503)~~
【分類號】：S816.3

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