Gudzenko E V, Varbanets L D, Kurchenko I M, Naconechnaya L T
Mikrobiol Z. 2016 Mar-Apr;78(2):33-42.
The aim of this work was to study α-L-rhamnosidase [КФ 3.2.1.40] - enzyme, which hydrolyse the terminal non-reduced α-1,2-, α-1,4- and α-1,6-linked L-rhamnose. As a result of screening conducted among 30 strains of micromycetes ability to synthesize α-L-rhamnosidase revealed in Penicillium tardum 60, 39, 2929, 2962, 2963, 2964, 2965, 2966, P. rugulosum 2778, 1652, 2766, P. restrictum 425, 2756, P. aculeatum 202, 217, 329, 2973, 2974, 2975, 2976, 2977, 2979 activity, which ranged from 0.07 to 0.53 OD/mg protein. The most active is brought out P. amleatum 202. From culture supernatant of this micromycete by fractionation with ammonium sulfate (90 % saturation) complex enzyme preparation was obtained and its physico-chemical properties were studied. It was shown that enzyme has pH optimum 3.0, thermooptimum - 60 °C and displayed stability in pH values from 2.0 to 4.0 during 90 min. At pH 5.0 the activity of complex enzyme preparation insignificantly decreased and appears to be up 40 % from initial one. At optimal pH value 3.0 and temperature 15 °C α-L-rhamnosidase tested was stable during 3 days. In addition to α-L-rhamnosidase enzyme preparation of P. arnleatum 202 exerted also β-D-glucosidase activity.
本研究旨在探究α-L-鼠李糖苷酶[EC 3.2.1.40]，该酶可水解末端非还原的α-1,2-、α-1,4-和α-1,6-连接的L-鼠李糖。在对30株微霉菌进行筛选的过程中，发现迟缓青霉60、39、2929、2962、2963、2964、2965、2966，皱折青霉2778、1652、2766，局限青霉425、2756，针刺青霉202、217、329、2973、2974、2975、2976、2977、2979具有合成α-L-鼠李糖苷酶的能力，其活性范围为0.07至0.53 OD/毫克蛋白质。活性最高的是针刺青霉202。通过硫酸铵分级分离（90%饱和度）从该微霉菌的培养上清液中获得了复合酶制剂，并对其理化性质进行了研究。结果表明，该酶的最适pH为3.0，最适温度为60℃，在pH值2.0至4.0的范围内90分钟内表现出稳定性。在pH 5.0时，复合酶制剂的活性略有下降，仍比初始活性高40%。在最适pH值3.0和温度15℃下，所测试的α-L-鼠李糖苷酶在3天内保持稳定。除α-L-鼠李糖苷酶外，针刺青霉202的酶制剂还具有β-D-葡萄糖苷酶活性。
