[真菌Penicillium sp. IB-37-2合成外-β-氨基葡萄糖酶]。

G E Aktuganov, N F Galimzyanova, G A Teregulova, A I Melentjev
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引用次数: 0

摘要

分离到一株对壳聚糖(SD ~ 80 ~ 85%)具有活性但对胶体几丁质活性较低的青霉b -37-2。在含有壳聚糖或蟹壳甲壳素的营养培养基中,在28°C、pH为3.5-7.0、220 rpm的条件下,真菌的壳聚糖酶生物合成水平较高(1.5-3.0 U/mL)。通过超滤和疏水色谱纯化青霉IB-37-2的壳聚糖酶复合物,再进行变性电泳,得到两个优势蛋白,分子量分别为89和41 kDa。纯化后的酶复合物在50-55°C和pH为3.5-4.0的条件下具有最大的活性(溶解壳聚糖的最大水解速率)和稳定性。该酶制剂还能水解层粘连蛋白、β-(1,3)-(1,4)-聚糖和胶体几丁质。利用青霉b -37-2分离物对壳聚糖进行外水解,得到单产物d -氨基葡萄糖。
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[Synthesis of exo-β-glucosaminidase BY FUNGUS Penicillium sp. IB-37-2].

A new strain Penicillium sp. IB-37-2, which actively hydrolyzes chitosan (SD ∼80–85%) but possesses low activity against colloidal chitin, was isolated. The fungus was observed to have a high level chitosanase biosynthesis (1.5–3.0 U/mL) during submerged cultivation at 28°C, with a pH of 3.5–7.0 and 220 rpm in nutrient media containing chitosan or chitin from shells of crabs. Purification of the chitosanase enzyme complex from Penicillium sp. IB-37-2 by ultrafiltration and hydrophobic chromatography, followed by denaturing electrophoresis, revealed two predominant proteins with molecular weights of 89 and 41 kDa. The purified enzyme complex demonstrated maximal activity (maximal rate of hydrolysis of dissolved chitosan) and stability at 50–55°C and a pH of 3.5–4.0. The enzyme preparation also hydrolyzed laminarin, β-(1,3)-(1,4)-glycan, and colloidal chitin. Exohydrolysis of chitosan by the preparation isolated from Penicillium sp. IB-37-2 resulted in the formation of single product, D-glucosamine.

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