CAJ | 학술논문

为获得康氏木霉( Trichodermakoningii)内切β-葡聚糖苷酶组分,并研究其酶学性质,通过硫酸铵盐析、DEAE Sepharose FF阴离子交换层析和Sephadex G-75凝胶过滤层析对康氏木霉GIMP3.444纤维素酶主要组分进行分离纯化,SDS-PAGE电泳检测蛋白质纯度和分子量,MALDI-TOFMS鉴定蛋白质种类。结果显示,从康氏木霉所产的纤维素酶系中分离纯化获得一种内切β-葡聚糖苷酶组分,相对分子质量为44600,酶的比活力为19.65 U/mg。该内切酶的最适反应pH值为4.5,最适反应温度为55℃。以羧甲基纤维素钠(CMC-Na)为底物时,酶的米氏常数(Km)为7.22 mg/ml。不同金属离子对酶活性影响不同,其中Ca2+(0.3 mmol/L)对酶的抑制作用较强,抑制了近35%的活力。通过MALDI-TOFMS鉴定及数据库分析,确定该酶为一种新的内切β-葡聚糖苷酶组分。
위획득강씨목매( Trichodermakoningii)내절β-포취당감매조분,병연구기매학성질,통과류산안염석、DEAE Sepharose FF음리자교환층석화Sephadex G-75응효과려층석대강씨목매GIMP3.444섬유소매주요조분진행분리순화,SDS-PAGE전영검측단백질순도화분자량,MALDI-TOFMS감정단백질충류。결과현시,종강씨목매소산적섬유소매계중분리순화획득일충내절β-포취당감매조분,상대분자질량위44600,매적비활력위19.65 U/mg。해내절매적최괄반응pH치위4.5,최괄반응온도위55℃。이최갑기섬유소납(CMC-Na)위저물시,매적미씨상수(Km)위7.22 mg/ml。불동금속리자대매활성영향불동,기중Ca2+(0.3 mmol/L)대매적억제작용교강,억제료근35%적활력。통과MALDI-TOFMS감정급수거고분석,학정해매위일충신적내절β-포취당감매조분。
To identify the composition of cellulase produced by Trichodermakoningii, ammonium sulfate precipitati-on, DEAE-sepharose FF anion exchange chromatography and sephadex G-75 gel filtration chromatography were used for purification of the cellulase. The molecular weight and purity were determined by SDS-PAGE, and the protein was identi-fied by MALDI-TOFMS. A new endo-β-glucanase was identified with its molecular weight of 44 600 and the specific activity of 19. 65 U/mg. The optimum pH and temperature for the endo-β-glucanase were 4. 5 and 55 ℃. The Km value was 7. 22 mg/ml (CMC-Na). Ca2+(0. 3 mmol/L) could inhibit the enzymatic activity significantly by 35%. The enzymatic proper-ties of the new endo-β-glucanase was distinct from other endo-β-glucanase. It could be used to was analyze the synergetic mechanism of cellulases.