该文对上海临港地区分离的1株产β-内酰胺酶的菌株的酶学性质进行研究,并初步提高了酶液保存稳定性。通过结合抗生素平板法初筛和碘量法(定性)复筛,获得产β-内酰胺酶的菌株。通过形态学、生理生化试验和16S rRNA序列分析确定该菌株为蜡状芽孢杆菌(Bacillus cereus),命名为Bacillus cereus B03。酶学性质研究表明,该酶的最适反应温度为40 ℃,最适反应pH值为7.0,该酶在4~20 ℃、pH 5.0~7.5内稳定性较好。Mg2+、Fe2+和Mn2+对酶有不同程度的激活作用,其中Mg2+对酶的激活作用最强。Cu2+、Al3+和Fe3+对酶有不同程度的抑制作用,其中Al3+对酶的抑制作用最强。K+、Ca2+、Zn2+对酶活力的影响作用不是很大。在无菌粗酶液中添加50 g/L NaCl或100 g/L甘油后,由原酶液的保存5 d剩余相对酶活力93%左右延长到了30 d,稳定性效果提升了6倍。菌株B03所产β-内酰胺酶的酶学性质初步研究表明,该酶具有进一步研究和应用的价值。
This study is to exploit the biochemical properties of β-lactamase producing strains isolated from Shanghai Lingang area. The β-lactamase-producing strain was isolated by combination of antibiotic and the iodometry (qualitative) screening. Using morphology characterization, physiological and biochemical tests and 16S rRNA sequencing analysis, the strain was identified as Bacillus cereus B03. The enzymatic properties test results indicated that the optimum reaction conditions are at 40 ℃, and pH 7.0. The enzyme stored in the solution had good stability within range of 4- 20 ℃ and pH 5.0-7.5. The ion Mg2+, Fe2+ and Mn2+ had different degree of activation on the enzyme and Mg2+ showed the strongest effect. Meanwhile, the ion Cu2+, Al3+ and Fe3+ had different degree of inhibitory effects and Al3+ showed the strongest effect. No obvious effects of K+, Ca2+ and Zn2+ on the enzyme activity were found. After adding 50 g/L NaCl or 100 g/L glycerol to the aseptic crude enzyme solution, the enzyme activity remained 93% for 30 days storage.
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