CAJ | 학술논문

从青霉Penicillium sp. C6菌株基因组DNA中克隆到一个新的16家族的葡聚糖酶基因,命名为bglc6(GenBank登录号:JX854434).其cDNA全长为1044 bp,包含N端前20个氨基酸的信号肽序列和一个16家族的结构催化域. bglc6编码的氨基酸序列与Grosmannia clavigera kw1407的假设葡聚糖酶蛋白序列最高一致性为64%.成熟蛋白BglC6在毕赤酵母中已成功表达,酶学性质分析表明,BglC6最适pH为4.5,最适温度为50℃,在酸性范围内具有良好的稳定性.以大麦葡聚糖酶为底物时,BglC6的比活、Km及Vmax值分别为253.8 U/mg,8.42 mg/mL,478.4μmol/min·mg.蛋白BglC6对大多数金属离子有较好的抵抗能力.因此,该葡聚糖酶在动物饲料行业领域中具有潜在的应用前景.
종청매Penicillium sp. C6균주기인조DNA중극륭도일개신적16가족적포취당매기인,명명위bglc6(GenBank등록호:JX854434).기cDNA전장위1044 bp,포함N단전20개안기산적신호태서렬화일개16가족적결구최화역. bglc6편마적안기산서렬여Grosmannia clavigera kw1407적가설포취당매단백서렬최고일치성위64%.성숙단백BglC6재필적효모중이성공표체,매학성질분석표명,BglC6최괄pH위4.5,최괄온도위50℃,재산성범위내구유량호적은정성.이대맥포취당매위저물시,BglC6적비활、Km급Vmax치분별위253.8 U/mg,8.42 mg/mL,478.4μmol/min·mg.단백BglC6대대다수금속리자유교호적저항능력.인차,해포취당매재동물사료행업영역중구유잠재적응용전경.
Bglc6, a glycoside hydrolase with 16 families’ glucanase gene, was cloned from Penicillium sp. C6. The full-length of cDNA was 1 044 bp, including a putative signal peptide of 20 residues and a catalytic domain with glycosyl hydrolase of 16 families. The deduced amino acid sequence showed the highest identity (64%) with the putative endo-1,3(4)-β-glucanase from Grosmannia clavigera kw1407. The mature albumen BglC6 was successfully expressed in Pichia pastoris. The enzymatic characteristics analysis showed that the optimum pH and temperature for the recombinant glucanase BglC6 was pH 4. 5 and 50℃. It exhibited good stability within acid pH range. When using barley β-glucan as substrate, the specific activity, Km and Vmax values were 253. 8 U/mg, 8. 42 mg/mL and 478. 4μmol/min·mg, respectively. BglC6 had better resistance to majority metal ions. Therefore, this glucanase possess potential application in animal feed industry.