Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China.
J Ind Microbiol Biotechnol. 2012 Jun;39(6):869-76. doi: 10.1007/s10295-012-1087-z. Epub 2012 Feb 22.
A novel endo-1,3(4)-β-D-glucanase gene (bgl16C1) from Penicillium pinophilum C1 was cloned and sequenced. The 945-bp full-length gene encoded a 315-residue polypeptide consisting of a putative signal peptide of 18 residues and a catalytic domain belonging to glycosyl hydrolase family 16. The deduced amino acid sequence showed the highest identity (82%) with the putative endo-1,3(4)-β-glucanase from Talaromyces stipitatus ATCC 10500 and 60% identity with the characterized β-1,3(4)-glucanase from Paecilomyces sp. FLH30. The gene was successfully overexpressed in Pichia pastoris. Recombinant Bgl16C1 constituted 95% of total secreted proteins (2.61 g l⁻¹) with activity of 28,721 U ml⁻¹ in a 15-l fermentor. The purified recombinant Bgl16C1 had higher specific activity toward barley β-glucan (12,622 U mg⁻¹) than all known glucanases and also showed activity against lichenan and laminarin. The enzyme was optimally active at pH 5.0 and 55°C and exhibited good stability over a broad acid and alkaline pH range (>85% activity at pH 3.0-7.0 and even 30% at pH 11.0). All these favorable enzymatic properties make it attractive for potential applications in various industries.
从青霉（Penicillium pinophilum）C1 中克隆并测序了一种新型内切-1,3(4)-β-D-葡聚糖酶基因（bgl16C1）。该 945bp 全长基因编码一个由 18 个残基组成的假定信号肽和一个属于糖苷水解酶家族 16 的催化结构域的 315 个残基多肽。推导的氨基酸序列与来自塔宾曲霉（Talaromyces stipitatus）ATCC 10500 的假定内切-1,3(4)-β-葡聚糖酶具有最高的同源性（82%），与已鉴定的来自棘孢木霉（Paecilomyces sp.）FLH30 的 β-1,3(4)-葡聚糖酶具有 60%的同源性。该基因在巴斯德毕赤酵母（Pichia pastoris）中成功过表达。重组 Bgl16C1 构成了总分泌蛋白的 95%（2.61 g l⁻¹），在 15 升发酵罐中的活性为 28,721 U ml⁻¹。纯化的重组 Bgl16C1 对大麦β-葡聚糖（12,622 U mg⁻¹）的比活性高于所有已知的葡聚糖酶，并且对松藻聚糖和昆布多糖也具有活性。该酶在 pH 5.0 和 55°C 下具有最佳活性，在较宽的酸碱 pH 范围内具有良好的稳定性（在 pH 3.0-7.0 下活性超过 85%，甚至在 pH 11.0 下仍有 30%的活性）。所有这些有利的酶学特性使其在各个行业中具有潜在的应用吸引力。
